Melanocytes

MELANOCYTES/MELANOGENESIS
Milestones in Melanocytes/Melanogenesis
Vincent J. Hearing1
1
Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA
Correspondence: Vincent J. Hearing, E-mail: hearingv@nih.gov

doi:10.1038/skinbio.2011.1
Melanocytes and their production of

melanin pigment (a process called melanogenesis) have important roles in
determining the physiology of human
skin. The amount and type of melanin
produced, as well as its eventual distribution in the epidermis, dramatically
affects visible color, which ultimately
determines the various functions of
the pigment, such as photoprotection.
Normal pigmentation is regulated by
4250 genes (as per the latest count), and
they function during the development,
migration, survival, proliferation, and
differentiation of melanocytes and their
precursors (melanoblasts). Disruptions
of skin pigmentation can occur at all
of these stages, and mutations in these
pigment-related genes typically lead

to characteristic pigmentary diseases.
Pigmentation in the lower species is
critical for survival (as camouflage),
thermal regulation, etc. In humans,
however, it is important not only for
cosmetic reasons but also to protect the
skin from UV damage and the subsequent risk of skin cancers. With respect
to susceptibility to skin cancers, melanocyte function seems to be important at
many levels in addition to the simple
sunscreen effect of the melanin pigment
per se. The chemical and enzymatic
processes involved in the synthesis of
different types of melanins have also
been gradually elucidated. The study of
genes involved in regulating mammalian
pigmentation has had a dramatic advantage owing to the visible effects of

mutations in these genes, and pigment
variants of mice, fish, birds, and other
species have been collected for centuries, providing a rich resource for understanding the complex nature of their
interactions. Combined with the dramatic advances in methods for gene
sequencing, and functional analysis, our
understanding of the complex regulation
of human skin pigmentationand its
disruptions in pigmentary diseaseshas
advanced rapidly.
TO CITE THIS ARTICLE
Hearing VJ (2011) Milestones in melanocytes/
melanogenesis. J Invest Dermatol 131: E1
ARTICLES
Milestone no. 1
Milestone no. 2
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Key Discoveries in Melanocyte DevelopmentAkinori Kawakami and David E. Fisher

Unpacking Human Evolution to Find the Genetic Determinants of Human Skin
PigmentationEllen E. Quillen and Mark D. Shriver
Determination of Melanin Synthetic PathwaysVincent J. Hearing
The Genetics of Human Pigmentary DisordersJonathan L. Rees
Molecular Aspects of TanningBarbara A. Gilchrest
The Genetics of VitiligoRichard A. Spritz
MILESTONES | CUTANEOUS BIOLOGY
NOVEMBER 2011
E1
Key Discoveries in Melanocyte Development

Akinori Kawakami1 and David E. Fisher1
1
Department of Dermatology, Cutaneous Biology Research Center, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA
Correspondence: David E. Fisher, E-mail: dfisher3@partners.org

doi:10.1038/skinbio.2011.2
Melanocytes are melanin pigment-producing cells. Mammalian melanocytes

are categorized as cutaneous (follicular and epidermal) and extracutaneous (e.g., choroidal, cochlear).
Epidermal melanocytes contribute to
photoprotection and thermoregulation
by packaging melanin pigment into
melanosomes and delivering them to
neighboring keratinocytes. Melanocytes are derived from the neural
crest that is a migratory multipotent
population that gives rise to multiple
cell lineages, including neurons, glial
cells, medullary secretory cells, smooth
muscle cells, and bone and cartilage
cells. Coat color mutants in different
species have been useful for identifying genes involved in melanocyte
development.
Embryonic transplantation experiments played a significant role in early
efforts to investigate melanocyte development. Rawles (1947) conducted a
series of elegant transplantation experiments, which revealed that melanocytes originate from the neural crest.
The investigator transplanted various
axial levels of the embryonic central
nervous system, adjacent tissues of the
somite and lateral plate, and limb-bud
regions, separately and in combination,
from various developmental stages of
black mouse embryos, to the coelom of
White Leghorn chick embryos. Only
tissues containing the neural crest or
cells migrating from the neural crest
were found to produce melanophores.
In addition, Rawles found that the
portions of embryo that produce pigment cells vary at different developmental stages of donor embryos.
The quail nucleus exhibits condensed heterochromatin, which could
serve as a marker to distinguish quail
E2
cells from chick cells. Teillet and Le

Douarin (1970) studied melanoblast
migration from the neural crest using
a quail-chick xenograft transplantation
model. The investigators transplanted
different axial levels of quail neural
tube and neural crest to White Leghorn
chick embryos. They found that (1) at
embryonic days E4 and E5, the transplanted quail cells localize mainly in
the mesenchyme; (2) at E6 when the
dermis and the epidermis are formed,
quail cells (melanoblasts) begin to
migrate into the epidermis; (3) at E9
quail cells (melanoblasts) increase their
cytoplasmic volume and start producing melanin pigments; (4) at E10 and
E11 quail cells (melanoblasts) localize
in the basal layer of epidermis and
become dendritic.
Cell type-specific markers are useful tools to study the development
of certain cell types. However, it is
challenging to identify these markers.
Steel et al. (1992) found Tyrp-2/Dct to
be a specific marker of melanoblasts,
the precursors of melanocytes. Using
in situ hybridization, the investigators
found that Tyrp-2 expression was
detectable in melanoblasts as early as
10 days post coitum.
The finding of a marker for early
melanoblasts enabled scientists to look
for the mechanisms of coat color
mutations. Steel and W mutants exhibited a white spotting color pattern.
W and Steel encode, respectively, a
receptor tyrosine kinase, Kit, and its
ligand, which is known by several
different names: steel factor, stem cell
factor, mast cell growth factor, and Kit
ligand. The mutation Steel-dickie (Sld)
is a deletion of its transmembrane and
cytoplasmic domains so that only a
secreted form of stem cell factor is
NOVEMBER 2011
produced. Steel et al. (1992) found

that the number of melanoblasts in (Sld/
Sld) mutants began to decrease at
around 11 days post coitum. They also
found that the melanoblasts caudal of
the optic vesicle failed to migrate
toward the vesicle. These results suggested that the cell surface form of stem
cell factor is important for both the
survival and migration of melanoblasts.
Wehrle-Haller and Weston (1995) performed in situ hybridization with Kit,
Tyrp-2, and stem cell factor probes in Sl
(null) and Sld mutants to examine the
early dispersal and fate of melanoblasts
in order to elucidate the function of
stem cell factor in more detail. They
concluded that soluble stem cell factor
is sufficient for responsive melanoblast
precursors to initiate their dispersal
onto the lateral migration pathway,
and that cell-bound stem cell factor
was necessary for the survival of
melanoblasts in the newly formed
dermal mesenchyme.
Dorsky et al. (1998) found that
cranial neural crest cells destined to
encode pigment cells were located
adjacent to the Wnt-1 and Wnt-3a
expression domain, whereas neurons
were far from the Wnt-expressing
domain in zebrafish. Most lateral
cells, which become neurons when
forcibly overexpressing an activated
form of b-catenin, adopted a pigment-cell fate. Conversely, when the
investigators overexpressed a mutant
form of Tcf-3 or a dominant-negative
Wnt-1 to inhibit Wnt signaling in
medial neural crest cells, the number
of pigment cells decreased dramatically. Thus, Dorsky et al. provided key
evidence that Wnt signaling plays an
essential early role in pigment cell
formation.
Mutations in both Ednrb and Edn3,

piebald-lethal (Ednrbs-l), and lethal
spotting (Edn3ls) exhibit severe melanocyte defects. To investigate the function of EDNRB signaling during
melanocyte development, Shin et al.
(1999) expressed or repressed Ednrb
expression using a tetracycline-inducible system. Their results indicated
several important points: (1) Ednrb is
required for melanocyte development
between E10 and E12.5; (2) Ednrb may
not be required for premigratory melanocytes; (3) Ednrb may be required for
the initiation of melanoblast migration
and/or for their survival; (4) Ednrb is not
required for postmigratory epidermal
proliferation, survival, and differentiation of melanoblasts.
MITF is a master transcriptional
regulator of the melanocyte lineage.
MITF mutations cause Waardenburg
syndrome type 2a, an autosomal dominant (heterozygous) condition that is
characterized by deafness and partial
depigmentation due to localized deficiencies of melanocytes. Mutations at
the mouse Mitf locus show melanocyte
defects in the skin, eyes, and inner ears.
MITFs transcriptional target genes include lineage survival factors (e.g.,
apoptosis regulators) as well as pigmentation-related genes (several of
which are implicated in albinism).
Yasumoto et al. (1994) found that MITF
specifically activates the transcription
of tyrosinase through a consensus DNA
motif E-box (CA[C/T]TG) in the melanocyte lineage. Hemesath et al. (1994)
found that Mitf bound the same E-box

as either a homodimer or a heterodimer
with TFEB, TFEC, or TFE3, and proposed recognition of these factors as
a distinct MiT transcription factor
family. MITF, TFEB, and TFE3 have also
been implicated as amplified or translocated oncogenes in a variety of
human malignancies, including melanoma, soft tissue sarcomas, and renal
carcinomas.
More recent studies have sought to
identify and functionally characterize
melanocyte stem cell populations.
Nishimura et al. (2002) identified melanocyte stem cells and their niche
using Dct-lacZ transgenic mice, which
carry the lacZ reporter under the
control of the Dct promoter, and
ACK2, a blocking c-Kit targeted monoclonal antibody. The investigators
found that melanocyte stem cells
reside in the bulge region of the hair
follicle (lower permanent portion
of the follicle) and fulfill the criteria
for stem cells: immature, slow cycling,
and self-maintaining, with an ability
to regenerate progeny on activation at
early anagen. Subsequently, Nishimura
et al. (2005) observed that agerelated hair graying is associated
with defective self-maintenance of
melanocyte stem cells and physiological aging/depletion of melanocyte stem cells caused by ectopic
pigmentation or differentiation within
the niche.
Melanoblasts have, for many
years, been thought to arise from the
HH 22
HH 24
dr
dorsal neural tube and migrate dorsolaterally between the dermamyotome

and the ectodermatome and ventrally
through the developing dermis to
their final destination, the basal
layer of the epidermis, and the hair
follicles.
However,
melanoblasts
may localize to the limbs without a
dermal migration. Adameyko et al.
(2009) conducted an elegant combination of experiments to address this
question and reassess the origin of
melanocytes; they provided compelling
new evidence that Schwann cell
precursors may serve as the cells of
origin for a major fraction of skin
melanocytes (Figure 1).
Our knowledge of melanocyte
development has been expanding
rapidly, but many important questions remain unanswered. How do
signals and transcription factors cooperate during melanocyte development? How do the cells surrounding
melanoblasts contribute to melanocyte development? How does the
epigenetic status change at different
stages of melanocyte development?
Are melanocytes of different locations functionally the same, and
may they be interchanged for clinical
benefit? Uncharacterized pigmentation mutants and new technologies,
such as chromatin immunoprecipitation sequencing and the melanocyte
lineage-specific targeted knockout
mice (e.g., TyrHCRE, DctHCRE), are
permitting ever-expanding answers to
these questions.
HH 27
2b
MITF+, Sox10+
dm
nt
Sox10+
Isll+
drg
2a
snv
2a
Figure 1. Migratory pathways of chick trunk neural crest cells, adapted from Cell 2009; 139:366379. Region 1: Melanoblasts (Mitf/Sox10 positive cells)
migrating within the dorsolateral pathway (the classic melanoblast migration route). 2a: Melanoblasts located near ventral spinal nerves. 2b: Melanoblasts
along nerves of dorsal rami. nt, neural tube; drg, dorsal root ganglion; dm, dermamyotome; dr, dorsal ramus; snv, ventral branch of the spinal nerve; HH,
Hamburger Hamilton stage.
NOVEMBER 2011
E3
CONFLICT OF INTEREST
The authors state no conflict of interest.

Kawakami A, Fisher DE (2011) Key discoveries in
melanocyte development. J Invest Dermatol 131:
E2E4
REFERENCES
Adameyko I, Lallemend F, Aquino JB et al. (2009)
Schwann cell precursors from nerve innervation
are a cellular origin of melanocytes in skin. Cell
139:36679
E4
nocyte development, defines a discrete transcription factor family. Genes Dev 8:277080
Nishimura EK, Granter S, Fisher DE (2005)
Mechanisms of hair graying: incomplete melanocyte stem cell maintenance in the niche. Science
307:7204
Nishimura EK, Jordan SA, Oshima H et al. (2002)
Dominant role of the niche in melanocyte stemcell fate determination. Nature 416:85460
Rawles ME (1947) Origin of pigment cells from
neural crest in the mouse embryo. Physiol Zool
20:24870
Dorsky RI, Moon RT, Raible DW (1998) Control of

neural crest cell fate by the Wnt signalling
pathway. Nature 396:3703
Shin MK, Levorse JM, Ingram RS et al. (1999)

The temporal requirement for endothelin receptor-B signalling during neural crest development.
Nature 402:496501
Hemesath TJ, Steingrmsson E, McGill G et al.

(1994) Microphthalmia, a critical factor in mela-
Steel KP, Davidson DR, Jackson IJ (1992)

TRP-2/DT, a new early melanoblast marker,
NOVEMBER 2011
shows that steel growth factor (c-kit ligand)

is a survival factor. Development 115:
11119
Teillet MA, Le Douarin N (1970) The migration of
pigmentary cells studies by the method of
heterospecific grafts of neural tube in bird
embryo. C R Acad Sci Hebd Seances Acad Sci D
270:30958
Wehrle-Haller B, Weston JA (1995) Soluble
and cell-bound forms of steel factor activity
play distinct roles in melanocyte precursor
dispersal and survival on the lateral neural
crest migration pathway. Development 121:
73142
Yasumoto K, Yokoyama K, Shibata K et al. (1994)
Microphthalmia-associated transcription factor
as a regulator for melanocyte-specific transcription of the human tyrosinase gene. Mol Cell Biol
14:805870
Unpacking Human Evolution to Find the Genetic

Determinants of Human Skin Pigmentation
Ellen E. Quillen1 and Mark D. Shriver2
1
Department of Genetics, Texas Biomedical Research Institute, San Antonio, Texas, USA and
University Park, Pennsylvania, USA
Department of Anthropology, Penn State University,
Correspondence: Mark D. Shriver, E-mail: mds17@psu.edu

doi:10.1038/skinbio.2011.3
How many genes determine variation

in human skin color? The apparent
simplicity of the question is belied by
the complex evolutionary history of our
species with regard to this trait. This
question was first addressed in a
seminal 1964 paper, On the inheritance of human skin colour, by
Harrison and Owen (1964). This work
is notable in (1) using reflectance
spectroscopy to quantify skin pigmentation; (2) focusing on population
samples with varying levels of genetic
admixture; and (3) using a genetic
approach to investigate evolutionary
questions.
Prior to the adoption of reflectometry-based measurements of skin pigmentation, researchers relied on
subjective assessments of pigmentation using reference tiles like those
produced by Felix von Luschen.
Although they have been widely used
for 50 years, these tiles result in
rough categorical estimates with large
inter-observer error (unpublished data)
unsuitable for quantitative genetic
analysis. Harrison and Owen, on the
other hand, measured reflected light
across the visible spectrum in relatively
unadmixed West Africans and Europeans, as well as in four admixed
groups (F1 hybrid, European backcross,
West African backcross, and F2 hybrid).
By using measurements that were
truly continuous and considering both
population means and variance, these
investigators were able to estimate the
quantitative genetic underpinnings of
this trait and, in doing so, established
the importance of admixed human
populations in studies of skin color.
Without this innovative approach to

measuring skin color in appropriately
identified admixed populations, many
of the recent advances in our understanding of the genetics of human skin
color would not have been possible.
Human skin color may be the most
rapidly evolving trait in our species,
with relatively closely related populations showing quite large differences in
average pigmentation levels. One study
has estimated the interpopulation variation in pigmentation at 88%, compared to 1015% for arbitrarily selected
genetic markers (Relethford, 2002).
Variation in both cellular structure
and biochemical pathways underlies
pigmentary differences among groups.
For example, while all humans have
approximately the same number of
melanosomes, the melanosomes of
darkly pigmented individuals contain
more melanin, are larger, and are
distributed singly instead of being
grouped into a membrane (Sturm
et al., 1998). Additional variation stems
from mutations in the many genes that
compose the pigmentation pathway,
including differences in tyrosinase
activity (encoded by the TYR gene),
which is the rate-limiting enzyme for
melanogenesis (Fuller et al., 2001).
In the past few years we have made
dramatic advances in our understanding of the genetic basis of normal
pigmentation variation (for reviews,
see Parra, 2007; Sturm, 2009). To date,
11 genes are known to influence skin
pigmentation levels within and between various human populations. Studies have focused largely on candidate
genes or admixture linkage in recently
NOVEMBER 2011
admixed populations (primarily with

West African/European or Indigenous
American/European parental populations) and genome-wide association
studies in non-admixed populations.
One exception is a genome-wide
association study in South Asiansa
population that has substantial old
admixture (Stokowski et al., 2007). As
Harrison and Owen realized, admixed
populations are essential for understanding pigmentation differences
among populations, since the alleles
at many of the functional loci are fixed
or nearly fixed. In the absence of
genetic variation, it is impossible to
map the effect of a locus.
Based on their partitioning of variance
using data from F1, backcrossed, and F2
individuals, Harrison and Owen predicted that a minimum of six to eight
genes underlie the skin color differences
between European and West African
populations. More recently, researchers
considering contemporary admixed
populations, which had undergone admixture for many more generations,
estimated that at least 10 genes contribute
to population-level differences between
European and West African populations
(Parra et al., 2004). At present, SLC24A5,
SLC45A2 (MATP), KITLG, OCA2, TYR,
and ASIP have been implicated (reviewed
in Sturm, 2009). Additional genes and
separate mutations likely are responsible
for the differences between Indigenous
Americans and Europeans. Ancestry informative markers in CYP19A1, MYO5A,
and SLC24A5 have shown admixture
linkage in Hispanic populations (Hoggart
et al., 2003). However, many more genes
must be identified to account for the full
E5
range of variation within and among all

human populations.
Through analysis in admixed populations, rather than through the more
widely used genome-wide association
study approach, a picture of the evolutionary genetic architecture of human
skin pigmentation is emerging and
proving to be quite intricate. Lamason
et al. (2005) identified a mutation in
SLC24A5 associated with the golden
phenotype in zebrafish. An amino-acid
substitution in the human homolog
(SLC24A5*Thr111), discovered by the
HapMap project, was found to have the
single largest effect of any known gene
on skin color differences between
Europeans and West Africans. Interestingly, this mutation, which is fixed
across most of Europe, is rare (B2.5%)
in East Asia, suggesting that the mutations responsible for lighter skin pigmentation are not shared across these
populations. SLC45A2 has also been
shown to have a pattern or variation
similar to SLC24A5, with the European
skin-lightening allele being largely restricted to Europe and not found in East
Asian populations (Norton et al., 2007).
This finding suggests that the mutation
rose to fixation in Europe relatively
recently, possibly under substantial
selective pressure. To determine if
the SLC24A5*Thr111 allele found in
European populations has a functional
effect on melanogenesis, Ginger et al.
(2008) inserted it into the DNA of
cultured melanocytes. The resulting modification in the trans-Golgi
network-associated protein NCKX5
showed both a substantial decrease in
cation exchange and downregulation
of melanin production. Based on these
observations, Ginger et al. hypothesize
that this mutation could change the
acidity in the trans-Golgi network,
thereby modifying the activity of tyrosine, or, based on the timing of
SLC24A5 expression during cell proliferation, it could modulate the trafficking of pre-melanosomes. These
functional and admixture-based studies
have been supplemented recently by
several studies of genomic selection at
pigmentation genes.
The distributions of SLC24A5 and
SLC45A2, in particular, and skin pigmentation more generally, show a
E6
MITF, EDN3
1
ASIP, BCN2, KITLG,
MLPH, RGS19
SLC24A5, MATP,
TYRP1, MYO5A,
DTNBP1, EDA,
OCA2
DCT, EGFR,
DRD2
West
African
East
Asian
4
North
European
ADAM17, DCT,
ADAMTS20, ATRN, MC1R,
LYST, OCA2, EDA, TYRP1
EGFR, DRD2
Figure 1. The evolutionary genetic architecture of skin pigmentation in three populations. Although
gene flow has occurred among human populations, differences in the allele frequencies at pigmentation
genes are observed among. The genes listed have demonstrated signatures of selection as (1) shared
among all humans, (2) shared between East Asian and European populations, (3) unique to East Asian
populations, (4) unique to European populations, and (5) unique to West African populations.
steeper cline than is found for most

human genes and phenotypes. This
raises questions about the types of
evolutionary forces that would give rise
to such a pattern. Harrison and Owen
were among the first to take a genetic
approach to this question by examining
the differences in skin reflectance
levels (and, by inference, the genes of
persons of known admixture). The
variation among these admixed individuals sheds light on the differences that
have evolved between West African
and European populations. Recently,
dozens of scans for selection across the
human genome have confirmed strong,
repeated selection on skin pigmentation
genes in nearly every population considered (see Figure 1). This evidence is
found more frequently in non-African
populations than in African populations. This finding may be associated
with the relatively recent colonization
of new environments by these populations, as older selection is often more
difficult to detect. In the Old World, a
few genes show selection in both East
Asian and European populations (e.g.,
KITLG), but there is also evidence of
independent evolution of light skin
pigmentation in European and East
Asian
populationsmeaning
the
shared phenotype does not result from
a shared ancestral mutation. This is
illustrated in the example of SLC24A5
and SLC45A2, both of which show
signatures of selection in Europe, but
not elsewhere. These selective events
mirror the migration of humans into
NOVEMBER 2011
Europe and Asia within the past 50,000

years. However, not all selective events
were recent. For example, patterns of
genetic variation at EDN3 support
selection on pigmentation in the common ancestors of all humans prior to
their divergence (McEvoy et al., 2006).
Some researchers have suggested that
this may reflect the initial darkening of
skin pigmentation in pre-human ancestors as they lost the fur that protects the
light skin of our chimpanzee cousins.
Among the most striking findings of
many of these studies is the magnitude
of the selection seen at skin pigmentation genes. A detailed description of the
evolutionary pressures influencing skin
pigmentation can be found in Jablonski
and Chaplin (2000). Briefly, most
research supports the hypothesis that
darker skin pigmentation evolved
possibly more than oncebecause it
reduces photolysis of folic acid, which
protects against neural tube defects and
is essential in DNA replication and
repair, among other functions. There is
more controversy surrounding the prevailing hypotheses for the adaptive
benefit of lighter skin pigmentation. In
a low UVR environment, less dermal
melanin allows for the production of
more vitamin D3 (needed for normal
skeletal development as well as proper
functioning of the immune and cardiovascular systems). However, there is
ongoing debate over the primacy of
vitamin D in contributing to differential
fitness (Robins, 2009; Chaplin and
Jablonski, 2010).
Despite the ongoing investigations

into the most likely pressures, what is
clear is that selection on skin pigmentation has been as strong as or stronger
than the selection on genes involved
in immunity, reproduction, and diet
all of which are essential for human
fitness. Several groups found marked
over-representations of pigmentation
genes among the strongest genomic
signatures of selection in Europeans
(Lao et al., 2006; Voight et al., 2006;
Myles et al., 2007). Pigmentation genes
are more than twice as likely to show
evidence of selection than randomly
selected genes in both Chinese and
European populations (Williamson
et al., 2007), and are significantly more
likely to have high FST, a measure of the
genetic distance between two populations, than randomly chosen regions
of the genome (Pickrell et al., 2009).
Taken together, the more than 25
pigmentation genes showing evidence
of selection suggest a history of selection on skin color that likely stretches
back to before the emergence of
the first anatomically modern Homo
sapiens.
Despite the many genes already
implicated in human skin color variation, a substantial number of the
differences among populations cannot
be explained. There are more than
350 putative pigmentation loci identified in mouse models and cataloged
in the IFPCS color genes database (http://
www.espcr.org/micemut/); what, if any,
role these genes play in the unexplained variation in human pigmentation
remains unknown. The size of this
database gives credence to the hypothesis that many rare variants may contribute to pigmentation variation in
humans. Identifying these rare variants
will only be possible with the increased
use of dense marker panels and sequencing to assess non-European and
admixed populations. Moreover, it is
essential that genome sequences be
associated with phenotypes. Identification of variation among populations or
individuals at known pigmentation

genes fails to demonstrate that this
variation manifests in phenotypic differences.
Unpacking the ramifications of human evolution in shaping the genetic
architecture and distribution of human
skin pigmentation requires accurate
and objective measurement techniques. Although self-report and subjective assessments may be sufficiently
informative to identify genes for some
traits (e.g., albinism, red hair, and blue
eye color), more complete quantitative
genetic analyses require measurements
of the skin using reflectometry at
specific wavelengths. As a larger fraction of the variation in human skin
color is distributed across rather than
within populations, admixed populations are an efficient means to discover
genes and to study their combined
effects. These components, employed
together in Harrison and Owen more
than 45 years ago, have shaped modern
research into the evolutionary genetics
of human skin pigmentation.
The authors state no conflict of interest.

Quillen EE, Shriver MD (2011). Unpacking
human evolution to find the genetic determinants
of human skin pigmentation. J Invest Dermatol
131: E5E7
REFERENCES
Chaplin G, Jablonski NG (2010) Human skin
pigmentation as an adaptation to UV radiation.
Proc Natl Acad Sci USA 107:89628
Fuller BB, Spaulding DT, Smith DR (2001)
Regulation of the catalytic activity of preexisting
tyrosinase in black and Caucasian human melanocyte cell cultures. Exp Cell Res 262:197208
Ginger RS, Askew SE, Ogborne RM et al. (2008)
SLC24A5 encodes a trans-Golgi network protein
with potassium-dependent sodium-calcium exchange activity that regulates human epidermal
melanogenesis. J Biol Chem 283:548695
Harrison GA, Owen JJT (1964) Studies on the
inheritance of human skin colour. Ann Hum
Genet Lond 28:2737
Hoggart CL, Parra EJ, Shriver MD et al. (2003)
Control of confounding of genetic associations in
NOVEMBER 2011
stratified populations. Am J Hum Genet 72:

1492504
Jablonski NG, Chaplin G (2000) The evolution of
human skin coloration. J Hum Evol 39:57106
Lamason RL, Mohideen M-L, Mest J et al. (2005)
SLC24A5, a putative cation exchanger, affects
pigmentation in zebrafish and humans. Science
310:17826
Lao O, de Gruijter JM, van Duijn K et al. (2006)
Signatures of positive selection in genes associated with human skin pigmentation as revealed
from analyses of single nucleotide polymorphisms. Ann Hum Genet 71:35469
McEvoy B, Beleza S, Shriver MD (2006)
The genetic architecture of normal variation in
human pigmentation: an evolutionary perspective
and model. Hum Mol Genet Spec No. 2:
R17681
Myles S, Somel M, Tang K et al. (2007)
Identifying genes underlying skin pigmentation
differences among human populations. Hum
Genet 120:61321
Norton HL, Kittles RA, Parra EJ et al. (2007)
Genetic evidence for the convergent evolution of
light skin in Europeans and East Asians. Mol Biol
Evol 24:71022
Parra EJ (2007) Human pigmentation variation:
evolution, genetic basis, and implications for
public health. Yearbook Phys Anthropol 50:
85105
Parra EJ, Kittles RA, Shriver MD (2004) Implications of correlations between skin color and
genetic ancestry for biomedical research. Nat
Genet 36:S54 S60
Pickrell JK, Coop G, Novembre J et al. (2009)
Signals of recent positive selection in a worldwide
sample of human populations. Genome Res
19:82637
Relethford JH (2002) Apportionment of global
human genetic diversity based on craniometrics
and skin color. Am J Phys Anthropol 118:
3938
Robins AH (2009) The evolution of light skin
color: role of vitamin D disputed. Am J Phys
Anthropol 139:44750
Stokowski RP, Pant PV, Dadd T et al. (2007) A
genomewide association study of skin pigmentation in a South Asian population. Am J Hum
Genet 81:111932
Sturm RA (2009) Molecular genetics of human
pigmentation diversity. Hum Mol Gen 18:R917
Sturm RA, Box NF, Ramsay M (1998) Human
pigmentation genetics: the difference is only skin
deep. Bioessays 20:71221
Voight BF, Kudaravalli S, Wen X et al. (2006) A
map of recent positive selection in the human
genome. PLoS Biol 4:e72
Williamson SH, Hubisz MJ, Clark AG et al. (2007)
Localizing recent adaptive evolution in the human
genome. PLoS Genet 3:e90
E7
Determination of Melanin Synthetic Pathways

Vincent J. Hearing1
1
Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA
Correspondence: Vincent J. Hearing, E-mail: hearingv@nih.gov

doi:10.1038/skinbio.2011.4
Visible pigmentation of the skin, hair,

and eyes depends primarily on the
presence of melanin(s) in those tissues.
Melanins are produced by specific cells
called melanocytes. Not only is the
type of melanin produced important,
but also its eventual distribution in the
tissue dramatically affects visible color,
which ultimately determines the functions of the pigment, such as photoprotection (Gilchrest, 2011). Clearly,
the specification, migration, and differentiation during development of melanocyte precursors (melanoblasts) in
specific patterns are essential for eventual pigmentation in adults (Kawakami
and Fisher, 2011). Following is a
synopsis of critical findings that have
led to our current understanding of
the biochemical pathways and melanogenic factors involved in melanin
synthesis.
The key enzyme involved in the
synthesis of all types of melanins
from the initial precursor tyrosine is
tyrosinase (EC 1.14.18.1). Tyrosinases
have been described in many species,
including mammals and lower animals,
plants, and even fungi; in fact, the
earliest observations of the catalytic
function of tyrosinase were made in
extracts of mushrooms (Bourquelot
and Bertrand, 1895), which are still
widely used today as a highly enriched
source of that enzyme. All tyrosinases
depend on the binding of copper for
their catalytic function (Lerner et al.,
1950; Lerch et al., 1986), although
their substrate specificities and physical
properties can differ dramatically depending on the species (Lerner et al.,
1951; Hearing et al., 1980). The ratelimiting initial step in the biosynthesis
of melanin was initially thought to be
the hydroxylation of tyrosine to L-3,4E8
dihydroxyphenylalanine (DOPA) and

its immediate subsequent oxidation
to DOPAquinone (DQ). In melanocytic cells, the DQ formed will be
spontaneously converted to an orangecolored intermediate known as DOPAchrome. In vitro, the DOPAchrome
will spontaneously lose its carboxylic
acid group to form 5,6-dihydroxyindole
(DHI), which can then further oxidize
and polymerize to form a dense, highmolecular-weight complex now known
as DHI-melanin. This was initially
reported by Raper (1926), and the
pathway was later refined by Mason
(1948); hence, the biosynthetic pathway is frequently referred to as the
RaperMason pathway. Throughout
the 1950s, 1960s, and 1970s, the collaborative research groups at Yale
(headed by AB Lerner) and Harvard
(headed by TB Fitzpatrick) played key
roles in defining the involvement of
tyrosinase in the human skin pigmentation (Fitzpatrick et al., 1950), how its
activities were confined to melanosomes and how those organelles developed (Seiji et al., 1961; Szabo et al.,
1969), and the disruptions that occurred in those processes in many skin
pigmentary diseases (Breathnach et al.,
1965; Kawamura et al., 1971; Lerner
and Nordlund, 1978; Rees, 2011;
Spritz, 2011). Those findings, plus the
training of many post-doctoral fellows
and clinicians in their groups, played a
major role in establishing research
centers in Asia, Europe, and the Americas, which still have a strong influence
on studies of skin pigmentation and
related pigmentary diseases. As summarized in Figure 1, recent revisions of
this melanogenic pathway have shown
that DOPA is not a distinct intermediate produced initially from tyrosine, but
NOVEMBER 2011
is in fact produced later in the pathway

owing to the paired reduction of DQ
(Riley, 1999), and that downstream
tyrosinase-related enzymes can rearrange the DOPAchrome to form a
carboxylated intermediate (DHI-2-carboxylic acid) known as DHICA, as
discussed below.
Analysis of the structures of the
high-molecular-weight polymers of
melanins depended on the development of new techniques to analyze
these intractable pigments, and gradual
progress was made in defining those
structures, initially by Nicolauss group
in Naples and Swans group in the
United Kingdom (Swan, 1963; Nicolaus et al., 1964). In working with
melanins found in nature, it quickly
became apparent that there were two
major types, the brownblack melanins
now collectively known as eumelanins,
and the yellowred melanins now
collectively known as pheomelanins.
Protas group in Naples took the lead in
defining the structure of pheomelanin
and the involvement of sulfur as
responsible for its unique color and
properties (Prota, 1980). Prota and
colleagues, as well as Ito and Wakamatsu in Japan (Ito et al., 1984),
developed a series of more sensitive
and specific assays for eumelanin and
pheomelanin intermediates that gradually formed the basis for our understanding of how they are formed in
melanocytes and how they are copolymerized in situ. The critical
role of sulfhydryl groups in reacting
immediately with DQ upon its formation to form various combinations
of cysteinylDOPAs and downstream
reactions of those intermediates, via
cysteinylDOPA-quinones and benzothiazine intermediates, to produce
Tyrosine
Melanosome
TYR
DOPAquinone
Cysteine
TYR
O2
DOPA
CycloDOPA
5SCD or 2SCD
DQ
DOPA
Eumelanin
polymer
CD-quinones
DOPAchrome
Pheomelanin
polymer
DCT
o-Quinoneimine
CO2
DHI
DQ
DHICA
O2
O2
DOPA
1,4-Benzothiazine intermediates
TYR or
TYRP1
Eumelanin
Pheomelanin
Figure 1. Scheme showing the melanogenic pathway that occurs within melanosome, leading to the production of eumelanin and/or pheomelanin.
Abbreviations used are as defined in the text. Adapted from Expert Review of Dermatology 2011;6:97108, with permission from Expert Reviews
(Kondo and Hearing, 2011).
pheomelanins were gradually defined

by the groups of Rorsman in Sweden,
Ito in Japan, Thody in the United
Kingdom, and Prota in Italy (Agrup
et al., 1979; Ito and Fujita, 1985; Thody
et al., 1991; Napolitano et al., 1994).
Studies characterizing the structural
and physical properties of melanins in
various tissues have also been led by
the groups of Simon in the United
States, dIschia and Zecca in Italy, and
Sarna in Poland (Sarna, 1992; Zecca
et al., 2001; Liu et al., 2005; Pezzella
et al., 2009). The determination to
produce eumelanin and/or pheomelanin is regulated physiologically, primarily by the melanocortin 1 receptor
(MC1R) as modulated by its opposing
ligands, melanocyte stimulating hormone (MSH) and agouti signaling
protein (ASIP); this is discussed in more
detail by Rees (2011).
Unexpectedly, in 1980, Pawelek
and colleagues reported the novel
finding that a biological factor produced in melanocytes was able to
prevent the spontaneous decarboxylation of DOPAchrome to DHI, which
led to the production of a more soluble
and lighter-colored carboxylated melanin, now known as DHICA-melanin
(Korner and Pawelek, 1980). There was
some initial controversy about this
point, since that activity could be

mimicked in vitro by various divalent
metal cations (Palumbo et al., 1987),
but with the advent of molecular
biology and cloning, the race to
clone the tyrosinase gene led indirectly
to the identification of two tyrosinase-related proteins (now known as
TYRP1 and DCT); one of those was
quickly shown to have the enzymatic
activity of DOPAchrome tautomerase
(Tsukamoto et al., 1992). Orlow and
colleagues then demonstrated that
the three tyrosinase-related melanogenic enzymes polymerized in a complex within melanocytes that facilitated
their physiological interactions (Orlow
et al., 1994).
Because of its critical role in pigmentation, and the disruptions in normal pigmentation expected to arise
from mutations in its encoding gene,
Kwon and colleagues initially cloned
the gene encoding tyrosinase (Kwon
et al., 1987) as well as another key
melanosomal protein now known to be
critically involved in melanosome structure, Pmel17 (Berson et al., 2001). As
noted above, cloning of the tyrosinase
gene led to the cloning of two other
closely related genes, and a number of
groups were instrumental in identifying human pigmentary diseases assoNOVEMBER 2011
ciated with each of those genes, most

notably by the laboratories of King and
Spritz in the United States (Getting and
King, 1994; Spritz, 1994) and later by
many other groups (reviewed in Hearing and Leong, 2005; Nordlund et al.,
2006). Interestingly, the four known
forms of oculocutaneous albinism result
from molecular lesions that disrupt the
function of tyrosinase: OCA1, the most
severe form, results from mutations in
the gene encoding tyrosinase itself,
but OCA2, OCA3, and OCA4 (slightly
milder forms) result from mutations
in genes that affect the processing
and trafficking of tyrosinase to melanosomes (P, TYRP1, and MATP, respectively) (Toyofuku et al., 2001; Costin
et al., 2003). Many other pigmentary
disorders affect other basic processes
in addition to their effects on melanocytes and melanin production, most of
them by virtue of disrupting intracellular trafficking pathways involved in
organelle biogenesis (including melanosomes) and/or the intracellular transport
or transfer of melanosomes to neighboring keratinocytes. Such disorders include HermanskyPudlak syndrome,
Griscelli syndrome, and ChediakHigashi syndrome. Interested readers are
referred to an actively curated web site
that lists pigment-related genes and
E9
associated diseases (http://www.espcr.

org/micemut/).
One final consideration that is now
becoming more widely accepted is the
synthesis and function of melanins in
less obscure tissues than the skin, hair,
and eyes. Melanocytes do occur, usually as minor populations, in a wide
variety of other tissues, and their
functions are currently a matter of
conjecture and study. Among those
other tissues are the inner ear, the
substantia nigra, the heart, and adipose
tissue. Evidence is accumulating that
the melanins play important protective
roles in those tissues (Brito and Kos,
2008; Zecca et al., 2008; Randhawa
et al., 2009). A recent article summarizes many of those studies and
provides hints of interesting directions
they might take in the future (Brenner
and Hearing, 2009).
Melanins play important roles in
human skin for cosmetic appearance,
detoxification, and photoprotection,
among other functions. In lower species, melanins play critical roles in
survival (e.g., in camouflage of prey
and predator, in thermal regulation in
amphibians, etc.) In humans, melanin
content (and thus visible pigmentation
of the skin) has a dramatic effect on
skins resistance to UV radiation damage, and the risk of skin cancers in
lighter skin is 30- to 40-fold higher than
in darker skin. The roles of melanins in
other tissues are still being studied and
will no doubt provide a wealth of
information about their various functions in the skin as well.
The author states no conflict of interest.
ACKNOWLEDGMENTS
This research was supported in part by the
Intramural Research Program of the NIH, National
Cancer Institute.

Hearing VJ (2011) Determination of melanin
synthetic pathways. J Invest Dermatol 131:
E8E11
REFERENCES
Agrup G, Hansson C, Rorsman H et al. (1979)
Intracellular distribution of DOPA and 5-S-cysteinylDOPA in pigment cells with minimal pigment
formation. Acta Dermatol Venereol Suppl
59:3556
E10
Berson JF, Harper DC, Tenza D et al. (2001)

Pmel17 initiates premelanosome morphogenesis
within multivesicular bodies. Mol Biol Cell
12:345164
Lerner AB, Fitzpatrick TB, Calkins E et al. (1950)

Mammalian tyrosinase; the relationship of
copper to enzymatic activity. J Biol Chem 187:
793802
Bourquelot E, Bertrand A (1895) Le bleuissement et le noircissement des champignons.

Comp Rend Soc Biol 2:5824
Lerner AB, Fitzpatrick TB, Calkins E et al. (1951)

Mammalian tyrosinase; action on substances
structurally related to tyrosine. J Biol Chem
191:799806
Breathnach AS, Fitzpatrick TB, Wyllie LMA

(1965) Electron microscopy of melanocytes in
human piebaldism. J Invest Dermatol 45:2837
Brenner M, Hearing VJ (2009) What are melanocytes really doing all day long y? Exp Dermatol
18:799819
Brito FC, Kos L (2008) Timeline and distribution
of melanocyte precursors in the mouse heart.
Pigment Cell Melanoma Res 21:46470
Costin GE, Valencia JC, Vieira WD et al. (2003)
Tyrosinase processing and intracellular trafficking
is disrupted in mouse primary melanocytes carrying the uw mutation: a model for oculocutaneous
albinism (OCA) type 4. J Cell Sci 116:320312
Fitzpatrick TB, Becker Jr SW, Lerner AB et al.
(1950) Tyrosinase in human skin: demonstration
of its presence and of its role in human melanin
formation. Science 112:2235
Getting WS, King RA (1994) Molecular basis of
oculocutaneous albinism. J Invest Dermatol
103:131S6S
Gilchrest BA (2011) Molecular aspects of tanning. J Invest Dermatol 131:E147
Hearing VJ, Ekel TM, Montague PM et al. (1980)
Mammalian tyrosinase. Stoichiometry and measurement of reaction products. Biochim Biophys
Acta 611:25168
Hearing VJ, Leong SPL (2005) From Melanocyte
to Melanoma: The Progression to Malignancy. 1st
ed. New York: Humana Press
Ito S, Fujita K (1985) Microanalysis of eumelanin
and pheomelanin in hair and melanomas by
chemical degradation and liquid chromatography.
Anal Biochem 144:52736
Ito S, Fujita K, Takahashi H et al. (1984)
Characterization of melanogenesis in mouse and
guinea pig hair by chemical analysis of melanins
and of free and bound DOPA and 5-S-cysteinylDOPA. J Invest Dermatol 83:124
Kawakami A, Fisher DE (2011) Key discoveries
in melanocyte development. J Invest Dermatol
131:E24
Kawamura K, Fitzpatrick TB, Seiji M (1971)
Biology of Normal and Abnormal Melanocytes.
Baltimore: University Park Press
Kondo T, Hearing VJ (2011) Update on the
regulation of melanocyte function and skin
pigmentation. Expert Rev Dermatol 6:97108
Korner AM, Pawelek JM (1980) DOPAchrome
conversion: a possible control point in melanin
biosynthesis. J Invest Dermatol 75:1925
Kwon BS, Haq AK, Pomerantz SH et al. (1987)
Isolation and sequence of a cDNA locus for human
tyrosinase that maps at the mouse c-albino locus.
Proc Natl Acad Sci USA 84:74737
Lerch K, Huber M, Scheider HJ et al. (1986)
Different origins of metal binding sites in binuclear copper proteins, tyrosinase and hemocyanin. J Inorg Chem 26:2137
NOVEMBER 2011
Lerner AB, Nordlund JJ (1978) Vitiligo what is it?

Is it important? J Am Med Assoc 239:11837
Liu Y, Hong L, Wakamatsu K et al. (2005)
Comparison of structural and chemical properties
of black and red human hair melanosomes.
Photochem Photobiol 81:13544
Mason HS (1948) The chemistry of melanin:
mechanism of the oxidation of dihydroxyphenylalanine by tyrosinase. J Biol Chem 172:8399
Napolitano A, Costantini C, Crescenzi O et al.
(1994) Characterization of 1,4-benzothiazine
intermediates in the oxidative conversion of 5S-cysteinyldopa to pheomelanins. Tetrahedron
Lett 35:63658
Nicolaus RA, Piattelli M, Fattorusso E (1964)
The structure of melanins and melanogenesis. IV.
On some natural melanins. Tetrahedron 20:
116372
Nordlund JJ, Boissy RE, Hearing VJ et al.
(2006) The Pigmentary System: Physiology and
Pathophysiology. 2nd ed. Edinburgh: Blackwell
Science
Orlow SJ, Zhou BK, Chakraborty AK et al. (1994)
High-molecular-weight forms of tyrosinase and
the tyrosinase-related proteins: evidence for a
melanogenic complex. J Invest Dermatol 103:
196201
Palumbo A, dIschia M, Misuraca G et al.
(1987) Effect of metal ions on the rearrangement
of DOPAchrome. Biochim Biophys Acta 925:
2039
Pezzella A, Iadonisi A, Valerio S et al. (2009)
Disentangling eumelanin black chromophore:
visible absorption changes as signatures of
oxidation state- and aggregation-dependent dynamic interactions in a model water-soluble 5,6dihydroxyindole polymer. J Am Chem Soc 131:
152705
Prota G (1980) Recent advances in the chemistry
of melanogenesis in mammals. J Invest Dermatol
75:1227
Randhawa M, Huff T, Valencia JC et al. (2009)
Evidence for the ectopic synthesis of melanin in
human adipose tissue. FASEB J 23:83543
Raper HS (1926) The tyrosinase-tyrosine reaction: production of L-3.4-dihydroxyphenylalanine
from tyrosine. Biochem J 20:73542
Rees JL (2011) The genetics of human pigmentary disorders. J Invest Dermatol 131:E123
Riley PA (1999) The great DOPA mystery: the
source and significance of DOPA in phase I
melanogenesis. Cell Mol Biol (Noisy-le-grand)
45:95160
Sarna T (1992) Properties and function of the
ocular melanina photobiophysical view. J
Photochem Photobiol 12:21558
Seiji M, Fitzpatrick TB, Birbeck MSC (1961) The
melanosome: a distinctive subcellular particle of
mammalian melanocytes and the site of melanogenesis. J Invest Dermatol 36:24352
Spritz RA (1994) Molecular genetics of

oculocutaneous albinism. Hum Mol Gen 3:
146975
Thody AJ, Higgins EM, Wakamatsu K et al. (1991)

Pheomelanin as well as eumelanin is present in
human epidermis. J Invest Dermatol 97:3404
Spritz RA (2011) The genetics of vitiligo. J Invest

Dermatol 131:E1820
Toyofuku K, Wada I, Valencia JC et al. (2001)

Oculocutaneous albinism (OCA) types 1 and 3 are
ER retention diseases: mutations in tyrosinase
and/or Tyrp1 influence the maturation, degradation of calnexin association of the other. FASEB J
15:214961
Swan GA (1963) Chemical structure of melanins.

Ann NY Acad Sci 100:100519
Szabo G, Gerald AB, Pathak MA et al.
(1969) Racial differences in the fate of
melanosomes in human epidermis. Nature
222:1081
Tsukamoto K, Jackson IJ, Urabe K et al. (1992)

A second tyrosinase-related protein, TRP2, is a
NOVEMBER 2011
melanogenic enzyme termed DOPAchrome tautomerase. EMBO J 11:51926

Zecca L, Bellei C, Costi P et al. (2008) New
melanic pigments in the human brain that
accumulate in aging and block environmental
toxic metals. Proc Natl Acad Sci USA 105:
1756772
Zecca L, Tampellini D, Gerlach M et al. (2001)
Substantia nigra neuromelanin: structure, synthesis, and molecular behaviour. J Clin Pathol Mol
Pathol 54:4148
E11
The Genetics of Human Pigmentary Disorders

Jonathan L. Rees1
1
Department of Dermatology, The University of Edinburgh, Edinburgh, UK
Correspondence: Jonathan L. Rees, E-mail: jonathan.rees@ed.ac.uk

doi:10.1038/skinbio.2011.5
Interest in the genetics of human and

animal pigmentation is longstanding.
Variation in human pigmentary form
of skin, hair, and eyesis one of the
most striking polymorphic human
traits. Ever since Charles Darwin, biologists have been asking how and why
did Nature fashion men so differently
across the earth? Moving beyond what
we might consider natural physiological variation in skin color to the
pathological, human pigmentary disorders such as albinism is often clinically
striking. Therefore, some syndromes
were described early in modern medical history, and what we would now
recognize as modern genetic insights
into their nature were present almost as
early as the rediscovery of Gregor
Mendels work just over a century ago.
The two decades spanning the start
of the twenty-first century have been
extraordinarily productive for those
interested in human pigment genetics.
Of course, the influence and power
of what was once called the New
Genetics has been felt across much
of modern biomedicine, but to this
skin-watcher, at least, few areas of
study have seen such progress with
apparently so modest an investment in
funding. Recounting the story of this
advance is necessary not merely to pay
homage to those who laid the foundations that later successes were built on,
but also as a lesson for those working in
other fields of biomedicine where the
problems often appear less tractable.
The development of the mouse
fancy, and the subsequent institutionalization of this in the twentieth century,
provided a fertile resource for those
keen to understand mammalian biology (Lamoreux et al., 2010). Interest in
this topic, however, was not limited to
E12
those, such as skin biologists or dermatologists, whose interests areprofessionally speakingskin deep, but
included those far-sighted biologists
who recognized that the astonishing
diversity of murine coat color mutants
offered a way to address the general
problem of how genes work. When
technological advances in molecular
genetics made mammalian gene identification almost routine, the resource
of the mouse fancy offered an unrivaled
experimental model system both for
those interested in human pigmentation
and for those whose goals were more
general.
PIGMENT GENES: MENDEL FIRST
In 1990, the genetic basis of oculocutaneous albinims 1 (OCA1), a common

form of albinism, was shown to be
attributable to mutations in the enzyme
tyrosinase (Giebel et al., 1990, 1991).
Subsequently, many of the other forms
of albinism and related disorders were
attributed to mutations in a range of
other genes: thus mutations at the P
locus caused OCA2 (Rinchik et al.,
1993); mutations of tyrosinase-related
protein (TRP-1), OCA3 (Manga et al.,
1997); and mutations of a membrane
transporter protein (MATP), OCA4
(Newton et al., 2001). Genes causing
the various types of Hermansky Pudlak
syndrome and Griscelli syndromes
were discovered, as well as the gene
underpinning the Chediak-Higashi syndrome (for reviews see Nordlund et al.,
2006). During the same period, the role
of the melanocortin 1 receptor (MC1R)
in causing the quasi-Mendelian trait of
red hair, as well as freckling and sun
sensitivity, was described (Valverde
et al., 1995; Flanagan et al., 2000).
Much of this work relied on prior
NOVEMBER 2011
experimental work in the mouse and

various degrees of extrapolation to man
(Robbins et al., 1993). More recently,
other model systems, notably the
zebrafish, have provided key insights
(Lamason et al., 2005). Finding a gene
does not mean that a disorder is
understood (let alone cured), but gene
identification accelerates future advances by providing a set of tools that
facilitate cell biology and physiology.
In turn, the clinician benefits from the
clinical bootstrapping gene identification allows, with a rationalization of
our understanding of the various syndromes and a fresh canvas on which to
sketch out relations between genotype
and phenotype.
PIGMENT GENES: GETTING MORE COMPLEX
Although often proposed as a research

strategy, there is no a priori reason why
genes involved in highly penetrant and
rare disorders should underpin normal
physiological variation. In the case of
human pigmentation, however, this
strategy has been remarkably successful, meaning that many of the genes
involved in the highly penetrant disorders such as albinism have proven to
be key determinants of physiological
variation in skin, hair, and eye color in
the general population. There are two
practical implications of this. First, such
loci contribute to variation in human
skin disease, particularly human skin
cancer. Hence, whereas red hair approximates to an autosomal recessive
trait, variants of the responsible gene
(MC1R) contribute to susceptibility to
sun sensitivity, most notably to skin
cancer risk (Smith et al., 1998;
Healy et al., 2000). Perhaps the
clearest example of this was work
showing that particular MC1R alleles
were determinants of the age of onset

of melanoma in families harboring p16/
CDKN2A mutations (Box et al., 2001),
and other interactions between MC1R
and OCA2 on melanoma incidence
(Duffy et al., 2004). Whereas the
Mendelian disorders allowed us to
cleave nature at the joints, facilitating
gene identification, what we learned
could then be used to study that more
subtle variations within the normal
population.
Second, the availability of largescale DNA analysis and genome-wide
scans, together with our existing
knowledge of the genes involved in
pigmentation, has opened up the possibility for robust testing of theories of
human evolution. Do we really believe
most variation in human pigmentary
characteristics is a result of selection, or
could neutral change (drift) account for
much of it? Is it just skin color that is
being selected for, or are other characteristics such as hair color also
important? Until the last two decades
it was simply not possible to rigorously
consider how we could test these
theories. Coupling large-scale DNA
analysis, knowledge of the many genes
contributing to human pigmentation,
and examination of diverse world populations through the Human Genome
Diversity-CEPH Panel, has moved the
study of human evolution from an
almost historical science to an experimental one in a way few could have
hoped for even in the mid 1980s
(Voight et al., 2006; Sabeti et al.,
2007; Pickrell et al., 2009; Pritchard
et al., 2010; Hancock et al., 2011).
It is of interest to ask why the study
of pigmentation genetics has been so
fruitful in comparison with the study
of other complex traits, such as the
inflammatory skin diseases. The importance of the mouse fancy cannot be
overstated, allowing the detection of
subtle and complex variation in coat
color, with our eyes still arguably
outperforming our biochemical or
cell-based assays. Similarly, and with
some pride to any dermatologist, our
ability to notice subtle and myriad
visual variations in form on human
skin and hair easily outperform any
machine-based assays. (Imagine a blind
observer trying to work out the genetics
of freckling based on measures of skin

cancer rates.)
Human pigmentary characteristics
have a high hereditability, and key
physiological components of the system
are either visible or easily demonstrable
(i.e., erythema). Finally, we have a
remarkably good understanding of
many aspects of human pigmentary
physiologydisorders such as albinism
and vitiligo coupled with the historic
geographic covariation of skin color
and skin cancer afford a coherent view
of the importance of pigmentation in
the ecological relation between sun and
skin color. And we know that this
interaction between ourselves and the
environment has been consistent over
longer periods of time than we can
safely say for infectious or inflammatory
disease, where changes in our diet and
social structures may have confounded
the relation between our genetic history
and modern phenotypes.
What of the future? It is clear that we
have not identified all the genes involved in human pigmentary variation.
We also do not have adequate quantitative models of how the known genes
interact. Nor can we say with confidence that we really understand the
fine details linking crude measures of
skin color with sensitivity to the effects
of ultraviolet radiation. What seems
clear, however, is that ever more
mining of our genome and how it
relates to our pigmentary diversity
across the earth will tell us more and
more about the human story.

Rees JL (2011) The genetics of human pigmentary disorders. J Invest Dermatol 131: E12E13
REFERENCES
Box NF, Duffy DL, Chen W et al. (2001) MC1R
genotype modifies risk of melanoma in families
segregating CDKN2A mutations. Am J Hum
Genet 69:76573
Duffy DL, Box NF, Chen W et al. (2004)
Interactive effects of MC1R and OCA2 on
melanoma risk phenotypes. Hum Mol Genet
13:44761
Flanagan N, Healy E, Ray A et al. (2000)
Pleiotropic effects of the melanocortin 1 receptor
(MC1R) gene on human pigmentation. Hum Mol
Genet 9:25317
NOVEMBER 2011
Giebel LB, Strunk KM, King RA et al.

(1990) A frequent tyrosinase gene mutation in
classic, tyrosinase-negative (type IA) oculocutaneous albinism. Proc Natl Acad Sci USA
87:32558
Giebel LB, Tripathi RK, Strunk KM et al. (1991)
Tyrosinase gene mutations associated with type
IB (yellow) oculocutaneous albinism. Am J
Hum Genet 48:115967
Hancock AM, Witonsky DB, Alkorta-Aranburu G
et al. (2011) Adaptations to climate-mediated
selective pressures in humans. PLoS Genet
7:e1001375
Healy E, Flannagan N, Ray A et al. (2000)
Melanocortin-1-receptor gene and sun sensitivity
in individuals without red hair. Lancet
355:10723
Lamason RL, Mohideen MA, Mest JR et al.
(2005) SLC24A5, a putative cation exchanger,
affects pigmentation in zebrafish and humans.
Science 310:17826
Lamoreux ML, Delamas V, Larue L et al. (2010)
The Colors of Mice. A Model Genetic Network.
Wiley-Blackwell: Chichester, UK
Manga P, Kromberg JG, Box NF et al.
(1997) Rufous oculocutaneous albinism in
southern African Blacks is caused by mutations
in the TYRP1 gene. Am J Hum Genet 61:
1095101
Newton JM, Cohen-Barak O, Hagiwara N et al.
(2001) Mutations in the human orthologue of the
mouse underwhite gene (uw) underlie a new form
of oculocutaneous albinism, OCA4. Am J of Hum
Genet 69:9818
Nordlund JJ, Boissy RE, Hearing VJ et al. (eds).
(2006) The Pigmentary System. Mass, USA:
Blackwell Publishing
Pickrell JK, Coop G, Novembre J et al. (2009)
Signals of recent positive selection in a worldwide
sample of human populations. Genome Res
19:82637
Pritchard JK, Pickrell JK, Coop G (2010) The
genetics of human adaptation: hard sweeps, soft
sweeps, and polygenic adaptation. Curr Biol
20:R20815
Rinchik EM, Bultman SJ, Horsthemke B et al.
(1993) A gene for the mouse pink-eyed dilution
locus and for human type II oculocutaneous
albinism. Nature 361:726
Robbins LS, Nadeau JH, Johnson KR et al.
(1993) Pigmentation phenotypes of variant
extension locus alleles result from point mutations that alter MSH receptor function. Cell
72:82734
Sabeti PC, Varilly P, Fry B et al. (2007) Genomewide detection and characterization of positive
selection in human populations. Nature
449:9138
Smith R, Healy E, Siddiqui S et al. (1998)
Melanocortin 1 receptor variants in an Irish
population. J Invest Dermatol 111:119
Valverde P, Healy E, Jackson I et al. (1995)
Variants of the melanocyte-stimulating hormone
receptor gene are associated with red hair and fair
skin in humans. Nat Genet 11:328
Voight BF, Kudaravalli S, Wen X et al. (2006) A
map of recent positive selection in the human
genome. PLoS Biol 4:e72
E13
Molecular Aspects of Tanning

Barbara A. Gilchrest1
1
Department of Dermatology, Boston University School of Medicine, Boston, Massachusetts, USA
Correspondence: Barbara A. Gilchrest, E-mail: bgilchre@bu.edu

doi:10.1038/skinbio.2011.6
Tanning is commonly understood as

increased melanization of the epidermis observed in skin following UV
exposure. It is further understood to
represent a host response that protects
against future UV-induced damage.
Although best studied in humans, tanning can also be observed in other
mammals and even in more primitive
animals such as sharks.
The molecular basis of tanning was
poorly understood well into the 1990s,
although UV irradiation of murine
melanoma cells had been shown to
increase the number of cell surface
receptors to a-melanocytestimulating
hormone (a-MSH) and to increase the
melanogenic response to a-MSH via
increased protein levels and activity of
tyrosinase, the rate-limiting enzyme
in melanin synthesis (Bolognia et al.,
1989; Chakraborty et al., 1991). However, the initiating molecular events
were unknown. Identification of
the responsible molecules ultimately
evolved from studies of the UV action
spectrum for critical events in human
skin, as well as from a philosophical
appreciation of tanning as a genome
protective response.
In the 1980s, investigators at Harvards Wellman Laboratories exposed
normal skin of volunteers to a wide
range of light doses using narrow band
sources across the UV spectrum and
observed them for several days to
determine the lowest dose at each
wavelength capable of producing a
delayed tan (Parrish et al., 1982). In
other experiments, they used the same
approach to define the action spectrum
for induction of cyclobutane pyrimidine dimers (CPDs), the most common
form of DNA damage following UV
exposure (Freeman et al., 1989). The
E14
spectra were virtually superimposable,

peaking at B300 nm, suggesting a
cause-and-effect relationship between
the immediate DNA damage and subsequent increased production and dispersion of epidermal melanin.
Eller et al. (1996) first documented
that agents acting exclusively on DNA,
such as DNA restriction enzymes,
stimulate melanin production in cultured pigment cells, at least in part
by increasing a-MSH binding and
the melanogenic response to a-MSH.
Although this did not exclude a role for
UV-mediated membrane effects, it did
directly implicate UV-mediated DNA
damage in the tanning response. Similarly, accelerating DNA damage repair
by providing UV-irradiated pigment
cells with T4 endonuclease V, the
bacterial enzyme that catalyzes the first
step in CPD resolution, also enhanced
the melanization response (Gilchrest
et al., 1993).
By the 1990s, DNA damage responses were recognized to be largely
mediated by the transcription factor
and tumor suppressor protein p53, also
termed the guardian of the genome
(Lane, 1992), and two groups independently asked whether p53 was involved
in mediating the tanning response.
Using p53 null osteosarcoma cells
transfected with wild-type p53, Nylander et al. (2000) showed that p53
activation increased read-out from a
transfected tyrosinase promoter linked
to a reporter gene, implying that p53
could directly or indirectly stimulate
tyrosinase transcription. This link between p53 and tanning was expanded
and refined using both human melanoma cells and a mouse model.
Compared with a p53-null parental
melanoma line and to the same line
NOVEMBER 2011
transfected with an empty vector,

melanoma cells transfected with wildtype p53 increased their tyrosinase
mRNA levels progressively over
72 hours following p53 activation
(Khlgatian et al., 1999; Khlgatian
et al., 2002). Confirming an earlier
report (Kichina et al., 1996), these
studies also showed an inverse relationship between total p53 protein (inactive) and tyrosinase levels. The
requirement for p53 activation to increase tyrosinase protein level and
epidermal melanin content in vivo
was confirmed by documenting the
tanning ability of wild type versus p53
knockout mice (Khlgatian et al., 2002).
The question remained whether p53
directly increased tyrosinase transcription, as no p53 consensus sequence
had been identified in its promoter
region.
The key observation that the UV
action spectrum for tanning is virtually
identical to that for CPD production,
had suggested that thymidine dinucleotide (abbreviated TT), the obligate
substrate for the majority of CPDs,
might itself serve as a molecular signal
for increased melanogenesis (tanning).
Using various models, the responses to
UV irradiation versus this DNA fragment were therefore compared. Indeed, TT increased mRNA and protein
expression of tyrosinase as well as
melanin content of cultured human
and murine pigment cells and of intact
guinea pig skin-containing melanocytes in the interfollicular epidermis,
in a time frame similar to that observed
after UV irradiation (Eller et al., 1994).
As well, in guinea pig skin, the
histological features were virtually
identical following either treatment:
increased total melanin with prominent
supranuclear melanin caps in the

keratinocytes, the familiar distribution
pattern for UV-tanned human skin, and
understood to maximally protect nuclear DNA from further UV damage.
Similar to UV irradiation, TT supplementation also increased cell surface
binding of a-MSH in cultured pigment
cells, presumptively through increased
a-MSH production by the cultured
pigment cells and/or increased binding
affinity for a-MSH to its cognate cell
surface receptor, the melanocortin 1
receptor (MC1R). In other experiments,
TT was observed to upregulate and
activate p53 in a variety of cell types
(Eller et al., 1997), bringing together the
two paths of investigation.
THE ROLE OF TELOMERE DISRUPTION
During the 1990s, initially outside of

the dermatological and pigment cell
biology communities, there was great
interest in the biological role of telomeres. Telomeres, the terminal portion
of all mammalian chromosomes, consist of short-tandem repeats of a noncoding DNA sequence, TTAGGG, in
all mammalian species. The de Lange
group observed that telomeres are not
linear but rather form a loop structure
and that disruption of this telomere
loop structure, exposing the singlestranded TTAGGG overhang, that is,
normally concealed within the proximal double-stranded telomere, leads
to DNA damage signaling (Karlseder
et al., 1999). These investigators determined that, depending on the experimental manipulation, loop disruption
led to activation of ataxia telangiectasia-mutated (Karlseder et al., 1999) or
ataxia telangiectasia-related (Denchi
and de Lange, 2007) kinases known to
mediate responses to DNA damage due
to gamma irradiation or UV, respectively, followed by activation of p53
and ultimately by apoptosis or senescence of the treated cells, depending
on cell type. Thus, experimental disruption of the telomere loop with
exposure of the TTAGGG repeat sequence rapidly led to the same cellular
responses as observed after acute DNA
damage due to UV (among other
injurious agents). This suggested that
introduction of DNA damage such as
CPDs, or influx of DNA repair proteins

to sites of such damage, might also
expose single-stranded TTAGGG repeats and, hence, that TTAGGG-containing DNA sequences might alone
suffice as a cellular DNA damage
signal (Gilchrest et al., 2009).
Interestingly, TT, one-third of the
repeat sequence, is the favored substrate for CPDs; guanine (GGG, onehalf the sequence) is the substrate for
almost all oxidative damage; and G
residues or adjacent adenineguanine
(AG) residues are the favored site for
formation of chemical adducts to DNA,
making telomeres an ideal target for
DNA damage (Gilchrest et al., 2009).
For example, it has been shown that
following UV irradiation, CPDs are
seven times more frequent in telomeric
DNA than in other portions of the
genome (Rochette and Brash, 2010). It
was therefore tempting to ask whether
TT was effective in stimulating melanogenesis precisely because it was a
part of the telomere repeat sequence
and whether full telomere repeat sequences would be even more effective.
Experiments demonstrated that an 11
base oligonucleotide GTTAGGGTTAG
was indeed far more effective on a
molar basis than TT in stimulating
tanning of cultured pigment cells or of
human skin supplemented ex vivo
(Gilchrest et al., 2009). Furthermore,
disrupting the telomere loop by the
same method as in the de Lange group
experiments also upregulated tyrosinase and tanned cultured human
melanocytes (Gilchrest et al., 2009).
Furthermore, telomere homolog oligonucleotides, termed T-oligos, increase
expression not only of tyrosinase, but
also of other melanogenic gene products such as TRP1, Mart1, and gp100
(Puri et al., 2004).
TYROSINASE ACTIVATION
It had long been recognized that the

rate of melanin production at baseline
and following UV exposure in either
cultured pigment cells or intact skin
depends on tyrosinase activity,
rather than simply on total tyrosinase
protein. Experiments by Park et al.
(1999) established that tyrosinase is a
phosphoprotein, active only when
NOVEMBER 2011
phosphorylated on serine residues in

its cytoplasmic domain that extends
beyond the melanosome membrane.
Only after this phosphorylation event,
mediated by protein kinase C-b (PKCb), does the intra-melanosomal portion
of tyrosinase catalyze the production of
eumelanin. Interestingly, PKC-b, which
is activated by diacylglycerol generated
from UV-irradiated cell membranes, is
among the gene products transcriptionally upregulated following UV irradiation (Park et al., 2006).
MELANOCYTEKERATINOCYTE INTERACTIONS
Tanning occurs to a far greater degree

in intact skin or in cocultures of
melanocytes or keratinocytes than in
isolated melanocyte cultures, suggesting that keratinocyte-derived products
contribute to the UV-induced tanning
response (Archambault et al., 1995).
Accordingly, many laboratories have
identified keratinocyte-derived gene
products that are upregulated by UV
irradiation and then act as paracrine
factors in the skin to stimulate survival,
melanogenesis, and/or melanin transfer
by melanocytes, including many
induced by p53 and/or T-oligos
(reviewed in Park et al., 2009).
Using an elegant mouse model in
which UV irradiation causes tanning
when the melanocytes express wildtype MC1R, but not when they express
loss-of-function MC1R variants associated in man with blond or red hair
and poor tanning ability, the Fisher
group demonstrated that the MC1R
ligand a-MSH, a cleavage product of
pro-opiomelanocortin (POMC), is regulated in keratinocytes by p53 via a p53
consensus sequence in the POMC gene
promoter (Cui et al., 2007). This
observation expanded the concept that
tanning is an integrated response of the
epidermis to DNA damage and identified the first direct target of p53
transcriptional activation involved in
melanogenesis. POMC production,
cleavage to a-MSH and other peptides,
upregulation by UV exposure, and
extracellular release had been documented in human keratinocytes by
several groups (Wintzen et al., 1996),
but the precise mechanism of UV
induction had been unknown. Whether
E15
the increased melanogenesis observed

in UV-irradiated cultured pigment
cells, in the absence of keratinocytes,
reflects POMC and subsequent a-MSH
upregulation by p53 in melanocytes
(an autocrine rather than paracrine
event) or the existence of additional
as yet unrecognized p53-regulated
genes involved in melanogenesis
remains to be determined (Park et al.,
2009).
Binding of keratinocyte-derived
a-MSH by wild-type MC1R on the
melanocyte surface results in increased
cyclic AMP production that in turn
leads to protein kinase A-mediated
transcriptional upregulation of the
microphthalmia transcription factor
(MITF; reviewed in Park et al., 2009).
MITF in turn transcriptionally upregulates the tyrosinase, TRP1, TRP2, and
PKC-b genes, increasing production of
melanin and specifically of the photoprotective brownblack eumelanin
polymer, highly capable of absorbing
UV photons that might otherwise
damage DNA (Park et al., 2009).
TANNING INCLUDES ENHANCED DNA
REPAIR CAPACITY
As noted above, it has long been

appreciated that tanning is the skins
major protective response against acute
and chronic UV damage. However, the
increased epidermal melanin content
associated with a good tan offers at
best a modest degree of photoprotection. For example, compared with
lightly pigmented epidermis, an excised darkly pigmented epidermis
placed over intact viable skin reduces
UV damage by at best a factor of 4
(Halder and Bridgeman-Shah, 1995).
This suggests that the tanning response
might consist of numerous changes
within the viable epidermis in addition
to enhanced melanogenesis. This speculation is made more appealing by the
recognition that p53 activation is a
critical initial tanning step, as described
above. Indeed, it was possible to
document that within 24 hours UV
irradiation of human or murine skin or
of cultured skin-derived cells resulted
in a 2- to 3-fold increase in the level of
several nucleotide excision repair proteins (Goukassian et al., 1999), many
E16
but not all of which are known to be

p53 regulated, as well as upregulation
of superoxide dismutases (SOD 1 and
2; Leccia et al., 2001), the enzymes
responsible for reduction of oxidative
damage due to UV or other insults. This
results in an accelerated rate of repair
for DNA damage following a subsequent UV exposure, compared with
that after an initial UV exposure (Arad
et al., 2007), as well as a reduction in
mutations and photocarcinogenesis
(Goukassian et al., 2004; Arad et al.,
2008). At least some of these effects
appear to be mediated through a-MSH
binding to MC1R (Kadekaro et al.,
2005), followed by induction of cAMP
(DOrazio et al., 2006; Passeron et al.,
2009) rather than through p53-stimulated direct upregulation of DNA repair
proteins. Thus, it seems that the UVinduced increase in epidermal melanin
content (conventionally termed tanning) is, but one aspect of a p53mediated DNA damage response that
includes upregulation of DNA repair
capacity and antioxidant defenses in
addition to an increased rate of melanogenesis, a protective adaptation
strikingly reminiscent of the SOS response first identified in bacteria (Eller
et al., 2008).
To date, the major molecular variations identified as impacting the tanning
response are MC1R polymorphisms
and splice variants (Rouzaud et al.,
2006; Cui et al., 2007). However, it
seems likely that variations in p53 and
other DNA damage response genes will
also be found to influence the tanning
response.

Gilchrest BA (2011) Molecular aspects of tanning. J Invest Dermatol 131: E14E17
REFERENCES
Arad S, Konnikov N, Goukassian DA et al. (2007)
Quantification of inducible SOS-like photoprotective responses in human skin. J Invest
Dermatol 127:262936
Arad S, Zattra E, Hebert J et al. (2008) Topical
thymidine dinucleotide treatment reduces
development of ultraviolet-induced basal cell
carcinoma in Ptch-1+/
mice. Am J Pathol
172:124855
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Archambault M, Yaar M, Gilchrest BA (1995)

Keratinocytes and fibroblasts in a human skin
equivalent model enhance melanocyte survival
and melanin synthesis after ultraviolet irradiation. J Invest Dermatol 104:85967
Bolognia J, Murray M, Pawelek J (1989) UVBinduced melanogenesis may be mediated through
the MSH-receptor system. J Invest Dermatol
92:6516
Chakraborty AK, Orlow SJ, Bolognia JL et al.
(1991) Structural/functional relationships between internal and external MSH receptors:
modulation of expression in Cloudman melanoma
cells by UVB radiation. J Cell Physiol 147:16
Cui R, Widlund HR, Feige E et al. (2007) Central
role of p53 in the suntan response and pathologic
hyperpigmentation. Cell 128:85364
DOrazio JA, Nobuhisa T, Cui R et al. (2006)
Topical drug rescue strategy and skin protection
based on the role of Mc1r in UV-induced tanning.
Nature 443:3404
Denchi EL, de Lange T (2007) Protection of
telomeres through independent control of ATM
and ATR by TRF2 and POT1. Nature
448:106871
Eller MS, Asarch A, Gilchrest BA (2008) Photoprotection in human skina multifaceted SOS
response. Photochem Photobiol 84:33949
Eller MS, Maeda T, Magnoni C et al. (1997)
Enhancement of DNA repair in human skin cells
by thymidine dinucleotides: evidence for a p53mediated mammalian SOS response. Proc Natl
Acad Sci USA 94:1262732
Eller MS, Ostrom K, Gilchrest BA (1996) DNA
damage enhances melanogenesis. Proc Natl Acad
Sci USA 93:108792
Eller MS, Yaar M, Gilchrest BA (1994) DNA
damage and melanogenesis. Nature 372:4134
Freeman SE, Hacham H, Gange RW et al. (1989)
Wavelength dependence of pyrimidine dimer
formation in DNA of human skin irradiated in
situ with ultraviolet light. Proc Natl Acad Sci USA
86:56059
Gilchrest BA, Eller MS, Yaar M (2009) Telomeremediated effects on melanogenesis and skin
aging. J Investig Dermatol Symp Proc 14:2531
Gilchrest BA, Zhai S, Eller MS et al. (1993)
Treatment of human melanocytes and S91
melanoma cells with the DNA repair enzyme
T4 endonuclease V enhances melanogenesis
after ultraviolet irradiation. J Invest Dermatol
101:66672
Goukassian DA, Eller MS, Yaar M et al. (1999)
Thymidine dinucleotide mimics the effect of solar
simulated irradiation on p53 and p53-regulated
proteins. J Invest Dermatol 112:2531
Goukassian DA, Helms E, Van Steeg H et al.
(2004) Topical DNA oligonucleotide therapy
reduces UV-induced mutations and photocarcinogenesis in hairless mice. Proc Natl Acad Sci
USA 101:39338
Halder RM, Bridgeman-Shah S (1995) Skin
cancer in African Americans. Cancer 75:66773
Kadekaro AL, Kavanagh R, Kanto H et al.
(2005) Alpha-melanocortin and endothelin-1
activate antiapoptotic pathways and reduce DNA
damage in human melanocytes. Cancer Res
65:42929
Karlseder J, Broccoli D, Dai Y et al. (1999) p53and ATM-dependent apoptosis induced by telomeres lacking TRF2. Science 283:13215
Khlgatian MK, Eller M, Yaar M et al. (1999)
Tyrosinase expression is regulated by p53.
J Invest Dermatol 112:548
Khlgatian MK, Hadshiew IM, Asawanonda P et al.
(2002) Tyrosinase gene expression is regulated
by p53. J Invest Dermatol 118:12632
Kichina J, Green A, Rauth S (1996) Tumor
suppressor p53 down-regulates tissue-specific
expression of tyrosinase gene in human melanoma cell lines. Pigment Cell Res 9:8591
Lane DP (1992) Cancer. p53, guardian of the
genome. Nature 358:156
Leccia MT, Yaar M, Allen N et al. (2001) Solar
simulated irradiation modulates gene expression
and activity of antioxidant enzymes in cultured
human dermal fibroblasts. Exp Dermatol 10:
2729
Nylander K, Bourdon JC, Bray SE et al.

(2000) Transcriptional activation of tyrosinase
and TRP-1 by p53 links UV irradiation to
the protective tanning response. J Pathol 190:
3946
Passeron T, Namiki T, Passeron HJ et al. (2009)

Forskolin protects keratinocytes from UVB-induced apoptosis and increases DNA repair
independent of its effects on melanogenesis.
J Invest Dermatol 129:1626
Park HY, Kosmadaki M, Yaar M et al.

(2009) Cellular mechanisms regulating human
melanogenesis. Cell Mol Life Sci 66:1493506
Puri N, Eller MS, Byers HR et al. (2004)

Telomere-based DNA damage responses: A new
approach to melanoma. FASEB J 18:137381
Park HY, Perez JM, Laursen R et al. (1999)

Protein kinase C-beta activates tyrosinase by
phosphorylating serine residues in its cytoplasmic domain. J Biol Chem 274:164708
Rochette PJ, Brash DE (2010) Human telomeres

are hypersensitive
to UV-induced DNA
Damage and refractory to repair. PLoS Genet
6:e1000926
Park HY, Wu C, Yonemoto L et al. (2006) MITF

mediates cAMP-induced protein kinase C-beta
expression in human melanocytes. Biochem J
395:5718
Rouzaud F, Costin GE, Yamaguchi Y et al. (2006)

Regulation of constitutive and UVR-induced skin
pigmentation by melanocortin 1 receptor isoforms. FASEB J 20:19279
Parrish JA, Jaenicke KF, Anderson RR (1982)

Erythema and melanogenesis action spectra of
normal human skin. Photochem Photobiol
36:18791
Wintzen M, Yaar M, Burbach JP et al.

(1996) Proopiomelanocortin gene product regulation in keratinocytes. J Invest Dermatol
106:6738
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E17
The Genetics of Vitiligo

Richard A. Spritz1
1
Human Medical Genetics Program and Departments of Pediatrics, Biochemistry and Molecular Genetics, and Craniofacial Biology, University of Colorado
School of Medicine, Aurora, Colorado, USA
Correspondence: Richard A. Spritz, E-mail: Richard.Spritz@ucdenver.edu
doi:10.1038/skinbio.2011.7
Vitiligo is one of the most striking of

all human disease phenotypes, and is
perhaps the most common pigmentary
disorder. The patchy loss of skin pigmentation and the marked contrast
between involved and uninvolved
skin particularly impacts persons of
color, with consequent stigmatization
that has long been recognized to result
in significant social inequality and
morbidity in affected individuals (Brito,
1885). Because of its visually evident
cutaneous manifestations, vitiligo has
been known for thousands of years.
However, only relatively recently has
there been real progress in understanding its pathobiological basis, which,
it is hoped, may facilitate progress in
vitiligo treatment and ultimately even
prevention. Surprisingly, in several
instances key discoveries seem to have
been overlooked or discounted, only
to be rediscovered and republished by
others a decade or more later.
Clinical studies of vitiligo remained
relatively primitive until the late 1950s,
when Lerner (1959) reported the first
systematic investigation of a large
patient series, suggesting the descriptive classification that is still essentially
in use today. In the treatment arena,
repigmentation of vitiligo by topical
or oral use of Ammi majus or Psoralia
corylifolia plant extracts or seeds,
particularly when combined with sun
exposure, was known in ancient Egypt,
India, China, and Japan, but was not
part of Western medical practice (Lindsay, 1932). It was not until after the
Second World War that specific treatment of vitiligo was pioneered by
Menon (1945) in India using UV light
and by El Mofty (1948) in Egypt using
the crystallized active components of
E18
Ammi majus, mainly 8-methoxypsoralen, both alone and in combination

with exposure to sunlight or UV light.
Despite extensive (and sometimes
fairly wild) speculation, the pathobiology of vitiligo remained completely
unknown until the next milestone, the
elegant demonstration by Hu et al.
(1959) that the affected skin of patients
with generalized vitiligo, the most
common form of the disorder, largely
lacked melanocytes, whereas melanocytes were present in both uninvolved
and repigmented skin of the same
patients. These same authors also
noted the transfer of pigment from
melanocytes to epithelial cells following UV irradiation, which was probably
the first recognition and description
of what later came to be called
the epidermal-melanin unit. These
observations led them to conclude
that successful treatment of vitiligo
therefore must depend on either
proliferation of residual functional
melanocytes within vitiligo lesions or
inward migration and proliferation of
melanocytes from surrounding normal
skin, as well as an intact ability of
epithelial cells to receive pigment from
active melanocytes.
Although the specific pathogenic
mechanism underlying melanocyte disappearance or destruction in vitiligo
remained obscure, the earliest clue to
an autoimmune origin, and perhaps the
earliest milestone in vitiligo, was the
report by Addison (1855) of a patient
with idiopathic adrenal insufficiency,
generalized vitiligo, and pernicious
anemia. The frequent concomitant
occurrence of multiple autoimmune
diseases, including generalized vitiligo,
was subsequently documented in many
NOVEMBER 2011
reports, particularly in that of Schmidt

(1926), and was later codified by
Neufeld and Blizzard (1980). In an
1872 lecture to the Silesian Society for
National Culture, Kobner, an eccentric
German dermatologist and mycologist,
described the isomorphe Reizeffekt
(Kobner, 1876), now known as the
Koebner phenomenon, the appearance of new, localized lesions at
sites of skin injury in psoriasis, which
was quickly extended to vitiligo and
many other skin disorders that have a
major autoimmune or autoinflammatory component (Hebra and Kaposi,
1874). Langhof et al. (1965) reported
autoantibodies to melanocyte components, and Gross et al. (1987) identified
perilesional infiltrates of cytotoxic
T-lymphocytes in patients with vitiligo.
There were also numerous case reports
of vitiligo developing in patients
with malignant melanoma, particularly
in those treated by immunotherapy
(Burdick and Hawk, 1964). Despite all
of these indicators, there was surprisingly enduring resistance to the acceptance of generalized vitiligo as an
autoimmune disease.
The role of genetic factors in vitiligo
was also considered early because of
the frequent clustering of cases among
close relatives (Stuttgen, 1950; Teindel,
1950), and eventual genetic epidemiological studies by Das et al. (1985)
supported multifactorial, polygenic
inheritance, which currently is termed
complex disease. Typical of the
1960s and 1970s, ABO, haptoglobin,
erythrocyte enzymes, and various
serum proteins were tested as genetic
markers for vitiligo, with negative
results. Beginning in the 1970s, a
plethora of analyses of HLA in vitiligo
were reported, with equivocal and

conflicting findings. Subsequently,
genetic association of vitiligo with many
other candidate genes was studied and
reported, though only PTPN22 and
perhaps CTLA4, both of which encode
immune regulators, are given high
credence today. The modern era of
genetic studies of generalized vitiligo
began indirectly, with the identification
of SLEV1 on chromosome 17p13, a
linkage signal in systemic lupus erythematosus families that also included
at least one relative with vitiligo (Nath
et al., 2001), underscoring a causal
genetic relationship between these two
diseases. SLEV1 was later identified as
NLRP1 (NALP1), a key regulator of the
innate immune system (Jin et al., 2007).
The most important recent vitiligo
developments were two large-scale
genomewide association studies of
generalized vitiligo, one in Caucasians
(Jin et al., 2010) and the other in
Chinese (Quan et al., 2010), which
together identified and confirmed at
least 16 different loci that contribute
to generalized vitiligo susceptibility. All
but one of these genes encode proteins
involved in regulation of the immune
system and/or have been genetically
associated with susceptibility to other
autoimmune diseases. The sole exception is TYR, encoding tyrosinase, the
key enzyme of melanin biosynthesis
and the principal vitiligo autoimmune
antigen. In Caucasians, a common TYR
missense variant, R402Q, confers both
relative protection from generalized
vitiligo and relative susceptibility to
malignant melanoma, by modulating
the presentation of the TYR peptide by
HLA-A2*01, thereby modulating recognition of melanocytes by the immune
system. These genes together account
for a relatively small fraction of the
genetic risk of generalized vitiligo,
indicating that many additional vitiligo susceptibility genes undoubtedly
remain to be discovered.
The results of the genetic studies
thus far show that generalized vitiligo is
a typical polygenic, multifactorial disorder, involving numerous different
susceptibility genes, and that the great
majority of these genes encode proteins
that regulate or mediate recognition or
destruction of melanocytes by the
immune system. Coupled with the

previous epidemiological and immunological evidence, there is no longer
any doubt that generalized vitiligo is a
complex autoimmune disease, some
genes determining general susceptibility to autoimmunity, and others
determining specific autoimmunity to
melanocytes. These findings begin to
highlight biological pathways that may
mediate the response of genetically
susceptible individuals to as-yet unknown environmental triggers of disease
onset and their response to factors that
mediate or modify its clinical course, as
well as their response to treatment.
Furthermore, recent evidence suggests that immune phenomena may
also contribute to the pathogenesis of
segmental vitiligo, a less common
form of the disorder, in which melanocyte loss remains quite localized,
often on the face, mostly occurring in
children and usually not associated
with other autoimmune diseases. Van
Geel et al. (2010) described a lymphocytic infiltrate of interferon-g-producing CD8 and some CD4 T cells
at the lesional margin in early segmental vitiligo, similar to those observed
in generalized vitiligo. Although occasionally segmental vitiligo and generalized vitiligo occur in the same families,
it is uncertain whether this represents
more than mere coincidence; thus, the
true pathobiological relationship between generalized vitiligo and segmental vitiligo remains to be elucidated.
This deeper understanding of vitiligo
pathogenesis allows us to anticipate
future milestones. Of course, it is
hoped that new insights into disease
pathogenesis and pathways will provide clues to new drugs or other therapeutic approaches. However, even in
the absence of new treatment modalities, such deeper understanding also
dictates a reconsideration of the current
approaches to vitiligo treatment. Vitiligo is a chronic autoimmune disease.
Melanocyte destruction remains ongoing even during attempts at treatment, possibly accounting, in part,
for the uneven therapeutic response
and long-term clinical course. It may
therefore prove optimal to investigate
combined approaches to treatment,
including both immunosuppressive
NOVEMBER 2011
and immunoregulatory approaches

that aim to reduce melanocyte destruction, while also including regenerative
approaches that aim to promote melanocyte repopulation of depigmented
regions. Likewise, an improved understanding of what vitiligo is also helps
clarify what it is not. The first Prime
Minister of India, Jawaharlal Nehru,
reportedly ranked vitiligo, leprosy, and
tuberculosis as the three most important medical problems in India (later
citations state malaria rather than
tuberculosis). Particularly in India, vitiligo and leprosy have long been confused, with a consequent inappropriate
stigmatization of vitiligo. In an effort to
combat this erroneous stigmatization,
on 27 December 2010, the Indian State
of Tamil Nadu issued a Government
Order that the terms ven kushtam
and ven kuttam, both meaning
white leprosy and often used as
equivalent to vitiligo, should be
abandoned. Truly, a milestone for
Indian vitiligo patients.

Spritz RA (2011) The genetics of vitiligo. J Invest
Dermatol 131: E18E20
REFERENCES
Addison T (1855) On the constitutional and
local effects of disease of the suprarenal
capsules. In: A Collection of the Published
Writing of the Late Thomas Addison, M.D.,
Physician to Guys Hospital, New Sydenham
Society, London, 1868. Reprinted in Med
Classics 1937; 2:24493
Brito PS (1885) On leucoderm, vitiligo, ven kuttam
(Tamil) or cabbare (Singhalese), and several new
methods of treatment. Br Med J 1:8345
Burdick KH, Hawk WA (1964) Vitiligo in a
case of vaccinia virus-treated melanoma. Cancer
17:720812
Das SK, Majumder PP, Majumdar TK et al.
(1985) Studies on vitiligo. II. Familial aggregation and genetics. Genet Epidemiol 2:25562
El Mofty AM (1948) A preliminary clinical report
on the treatment of leukoderma with Ammi majus
Linn. J Roy Egyptian MA 31:6515
Gross A, Tapia FJ, Mosca W et al. (1987) Mononuclear cell subpopulations and infiltrating
lymphocytes in erythema dyschromicum perstans
and vitiligo. Histol Histopathol 2:27783
Hebra F, Kaposi M (1874) On Diseases of the
Skin, Including the Exanthemata (Tay W, trans. and
ed.)., vol. III. London: New Sydenham Society
E19
Hu F, Fosnaugh RP, Lesney PF (1959) In vitro

studies on vitiligo. J Invest Dermatol 33:26780
Lindsay HCL (1932) Leukodermaits treatment.

Cal West Med 37:3641
susceptibility loci at 6q27 and the MHC. Nat

Genet 42:6148
Jin Y, Birlea SA, Fain PR et al. (2010) Variant of

TYR and autoimmunity susceptibility loci in generalized vitiligo. New Engl J Med 362:168697
Menon AN (1945) Ultra-violet therapy in cases of

leukoderma. Ind Med Gaz 80:6124
Schmidt M (1926) Eine biglanduiare Erkrankung

(Nebennieren und Schilddruse) bei Morbus
Addisonii. Verh Dtsch Ges Pathol 21:21221
Jin Y, Mailloux CM, Gowan K et al. (2007) NALP1

in vitiligo-associated multiple autoimmune
disease. New Engl J Med 356:121625
Kobner H (1876) Zur aetiologie der psoriasis.
Vjschr Dermatol 8:55961
E20
Nath SK, Kelly JA, Namjou B et al. (2001) Evidence

for a susceptibility gene, SLEV1, on chromosome
17p13 in families with vitiligo-related systemic
lupus erythematosus. Am J Hum Genet 69:14016
Langhof H, Feuerstein M, Schabinski G (1965)

Melaninantikorperbildung bei vitiligo. Hautarzt
16:20921
Neufeld M, Blizzard RM (1980) Polyglandular

autoimmune diseases. In: Symposium on Autoimmune Aspects of Endocrine Disorders (Pinchera
A, Doniach D, Fenzi GF et al., eds). New York:
Academic Press, 35765
Lerner AB (1959) Vitiligo. J Invest Dermatol

32:285310
Quan C, Ren YQ, Xiang LH et al. (2010) Genomewide association study for vitiligo identifies
NOVEMBER 2011
Stuttgen G (1950) Die Vitiligo in erbbiologischer

Betrachtung. Z Haut Geschlechtskr 9:4517
Teindel H (1950) Familiare Vitiligo. Z Haut
Geschlechtskr 9:45762
van Geel NA, Mollet IG, De Schepper S et al.
(2010) First histopathological and immunophenotypic analysis of early dynamic events in
a patient with segmental vitiligo associated
with halo nevi. Pigment Cell Melanoma Res 23:
37584

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MELANOCYTES/MELANOGENESIS

Correspondence: Vincent J. Hearing, E-mail: hearingv@nih.gov

Melanocytes and their production of

pigment-related genes typically lead

pigmentation has had a dramatic advantage owing to the visible effects of

Key Discoveries in Melanocyte DevelopmentAkinori Kawakami and David E. Fisher

MILESTONES | CUTANEOUS BIOLOGY

Key Discoveries in Melanocyte Development

Correspondence: David E. Fisher, E-mail: dfisher3@partners.org

Melanocytes are melanin pigment-producing cells. Mammalian melanocytes

cells from chick cells. Teillet and Le

produced. Steel et al. (1992) found

MILESTONES | CUTANEOUS BIOLOGY

Mutations in both Ednrb and Edn3,

found that Mitf bound the same E-box

dorsal neural tube and migrate dorsolaterally between the dermamyotome

MILESTONES | CUTANEOUS BIOLOGY

TO CITE THIS ARTICLE

Dorsky RI, Moon RT, Raible DW (1998) Control of

Shin MK, Levorse JM, Ingram RS et al. (1999)

Hemesath TJ, Steingrmsson E, McGill G et al.

Steel KP, Davidson DR, Jackson IJ (1992)

shows that steel growth factor (c-kit ligand)

MILESTONES | CUTANEOUS BIOLOGY

Unpacking Human Evolution to Find the Genetic

Department of Anthropology, Penn State University,

Correspondence: Mark D. Shriver, E-mail: mds17@psu.edu

How many genes determine variation

Without this innovative approach to

admixed populations (primarily with

range of variation within and among all

steeper cline than is found for most

Europe and Asia within the past 50,000

Despite the ongoing investigations

MILESTONES | CUTANEOUS BIOLOGY

individuals at known pigmentation

TO CITE THIS ARTICLE

stratified populations. Am J Hum Genet 72:

Determination of Melanin Synthetic Pathways

Correspondence: Vincent J. Hearing, E-mail: hearingv@nih.gov

Visible pigmentation of the skin, hair,

dihydroxyphenylalanine (DOPA) and

is in fact produced later in the pathway

pheomelanins were gradually defined

point, since that activity could be

ciated with each of those genes, most

associated diseases (http://www.espcr.

TO CITE THIS ARTICLE

Berson JF, Harper DC, Tenza D et al. (2001)

Lerner AB, Fitzpatrick TB, Calkins E et al. (1950)

Bourquelot E, Bertrand A (1895) Le bleuissement et le noircissement des champignons.

Lerner AB, Fitzpatrick TB, Calkins E et al. (1951)

Breathnach AS, Fitzpatrick TB, Wyllie LMA

Lerner AB, Nordlund JJ (1978) Vitiligo what is it?

MILESTONES | CUTANEOUS BIOLOGY

Spritz RA (1994) Molecular genetics of

Thody AJ, Higgins EM, Wakamatsu K et al. (1991)

Spritz RA (2011) The genetics of vitiligo. J Invest

Toyofuku K, Wada I, Valencia JC et al. (2001)

Swan GA (1963) Chemical structure of melanins.

MILESTONES | CUTANEOUS BIOLOGY

Tsukamoto K, Jackson IJ, Urabe K et al. (1992)

melanogenic enzyme termed DOPAchrome tautomerase. EMBO J 11:51926

The Genetics of Human Pigmentary Disorders