Nucleus:

Nucleus (L., nux, nut) is the most important part of the cell situated in the cytoplasm. All the
cellular activities are controlled by it. Nucleus is a directing and organizing unit without which
the cell could not exist. It was discovered by Robert Brown (1831) in flowering plants and is
now recognized as the structure that contains the hereditary material of the cell. The study of
nucleus or karyosome constitutes karyology (Gr., Karion, and nucleus).

Image Curtsey: http://2.bp.blogspot.com/Banswer.jpg
Position:
The location of nucleus varies in the cell depending upon the species. Usually it is situated in the
centre of the cell surrounded on all sides by cytoplasm. In green algae, Acetabularia, it shows
various positions, though mainly present in the basal part of cell. Generally the nuclei are
scattered in the cytoplasm.
Morphology:
1. Shape:
The shape of nucleus is variable according to cell type. It is generally spheroid but ellipsoid or
flattened nuclei may also occur in certain cells. The nuclear margins are generally smooth but
they may be lobulated bearing small infoldings of nuclear membrane as in leucocytes. In certain
white blood corpuscles the nucleus is dumbbell-shaped and exhibits variation during life history
stages. In human neutrophil, it is trilobed.
2. Number:
Mostly cell contains a single nucleus, known as mononucleate cell. Cells containing two nuclei
are known as binucleate cells (e.g., Paramecium), and cells of cartilage and liver. Sometimes
more than two nuclei (3 to 100 nuclei) are present in a single cell. Such cells are called
polynucleate cells.

Such cells in animals are called syncytial cells (e.g., osteoblast) and such plant cells are termed
coenocytes (e.g., siphonal algae). Cells having distinct nucleus are called eukaryotic types,
whereas cells without definite nucleus are called prokaryotic cells, (e.g., bacteria, etc.). The latter
possess scattered chromatin material (DNA) in the cytoplasm.
The mammalian erythrocytes (eukaryotic cells) possess no nuclei. In certain algae, like
Vaucheria and some fungi (e.g., black bread mold, Rhizopus nigricans) there occur numerous
nuclei in a single cell which is called a coenocyte. In animal cells, similarly, many nuclei may
occur in a single cell, as in malarial parasite, forming a syncytium. The striated muscle fibres are
good example of syncytium.
3. Size:
Size of nucleus is not constant and is generally correlated with DNA contents. The nuclear size is
a function of chromosome number, i.e., the size is variable depending upon the number of
chromosomes (DNA contents). Usually, there exists a ratio between the nuclear volume (Nv) and
the volume of the cytoplasm (Cv) for each cell type. It can be expressed by an equation called
nucleoplasmic index (NP) given by R. Hertwig which is as follows:
NP = Nv / Cv Nv
This index is fixed for each cell and any disturbance of equilibrium of NP acts as stimulus for
cell division. Cells containing more than one set of chromosome are called diploid or polyploid
cells, having larger number of chromosomes.
Thus, cells with more than the diploid number of chromosomes have usually larger nuclei. The
nucleus of resting period, when it does not undergo any division, is called the metabolic nucleus,
while dividing nucleus, is called kinetonucleus or arbeitonukleus in German language.

The nucleus consists of the following main parts: (1) Nucleolemma or nuclear membrane
(karyotheca) (2) Nuclear sap or karyolymph or nucleoplasm (3) Chromatin network or
fibres (4) Nucleolus (5) Endosomes!
[I] Nuclear membrane (karyotheca):

The nucleus is separated from the cytoplasm by a limiting membrane called as karyotheca or
nuclear membrane. This membrane plays an important role for the transport of the material
between the nucleus and the cytoplasm. Nuclear envelope regulates nucleocytoplasmic
exchanges and coordinates gene action with cytoplasmic activity.
It has direct connections with the endoplasmic reticulum and during cell division, this nuclear
envelope (nuclear membrane) develops as an extension of the endoplasmic reticulum applied to
the nucleus and subsequently modified. In the end of mitosis, i.e, in telophase, the cisternae of
the endoplasmic reticulum gather around the chromosomes and by fusing together they form the
nuclear membrane.
Structure:
The nuclear membrane appears to be a double membrane having interruptions or pores at
intervals. The outer one is called ectokaryotheca and inner one is termed endokaryotheca. Each
of the membranes is about 75 to 90 A thick and both the membranes enclose an intervening
space of about 100-150 A (Robertis et al., 1970), or 100 to 300 A (Cohn, 1970) or 400 to 700 A
(Burke, 1970), called as perinuclear space or cisterna.
Outer nuclear membrane is comparatively thicker than the inner membrane and has a rough
outline due to the presence of attached RNA particles which may form spirals, parallel lines or
crescents. The inner membrane is smooth having no ribosomes. The nuclear membrane is
followed by a supporting membrane, the fibrous lamina, of uniform thickness (300 A thick).
Nuclear pores:
The nuclear membrane possesses a number of nuclear pores or annuli, which vary from 40 to
145 per square micro-meter in nuclei of various plants and animals. Watson (1959) stated the
number of pores in mammalian cells as 10 percent of the total surface of the nucleus. In
amphibian oocytes, certain plant cells and protozoa the surface occupied by the pores may be as
high as 20 to 36 percent.
The nuclear pores are octagonal in shape, their diameter varies from 400-1000 A, and they are
separated from each other by a space of 1500 A. The nuclear pores are enclosed by circular
annuli. At the annulus the inner and outer membranes of the nuclear envelope fuse.
The pores and annuli collectively form the pore complex. Eaqh annulus consists of eight
granules of about 15 nm, which are present on both the nuclear and cytoplasmic surfaces. Inside
the pore is a central granule. Fine fibrils (about 30 A in diameter) extend from central granule to
the peripheral granules, forming a cartwheel structure.
A less defined amorphous annular material is present in the opening itself. This material is
digested by trypsin and remains unaffected when exposed to ribonuclease and
deoxyribonuclease. It means that annular material is protein in nature. The pore complex is a
rigid structure present in a fixed number according to cell type. In certain physiological stages,
however, they may change in number.


For example, they are reduced in number in maturing erythroblasts and spermatids and it is due
to low transcriptional activity of these cells. In some cases, the pore complex is covered by a thin
membrane. It has been suggested that the annulus may serve as a sphincter, alternately
decreasing and increasing the size of the pore with varying conditions.
Some evidence suggests the presence of myosin in the annulus area (Du Praw, 1970). Annulus is
supposed to be a hollow cylinder fitting into the nuclear pore (Witschnitzer, 1958). The lumen of
the cylinder is 500 A in diameters representing the nuclear pore.
The wall of the cylinder consists of eight evenly placed microtubules or microcylinders. Each
microtubule is about 200 A in diameters. According to Viviers, a central microtubule of 150 to
180 A diameters is present with in the lumen of annulus and is attached to its inner wall by
fibrous struts.


Amorphous annular material extends beyond the pore margins (Franke and co-workers, 1966-
74). The materials exchanged between nucleus and cytoplasm must traverse the nuclear pore
complexes. Thus, annuli or pores control the passage of some molecules and particles, even
some ribosome components, between nucleus and cytoplasm. This exchange is very selective
and allows passage of only certain molecules of either low or very high molecular weight. The
nuclear envelope is a diffusion barrier for ions as small as K
+
, Na
+
or l
-
.
On the other hand, very large structures such as ribosomal subunits, which are assembled in the
nucleolus, are able to leave the nucleus through the nuclear pore complexes. The unit membranes
of karyotheca are composed of protein and lipid, like plasma membrane.
At the margins of these pores, the two unit membranes are continuous and at certain places this
nuclear membrane joins the membrane of endoplasmic reticulum. In the apothecial
(reproductive) cells of Mollisia (an ascomycetes fungus), the nuclear membrane makes direct
contact with the plasma membrane of cell.
[II] Nuclear sap (karyolymph or nucleoplasm):
The nucleus contains a transparent, semi-solid, granular and homogeneous matrix during
interphase called as nuclear sap or karyolymph (enchylema). This karyolymph is a fluid
substance containing many particles and network. Primarily it is composed of proteinous
material and is the main site for enzyme activity. This nuclear sap also shows variable
appearance during different stages of cell division. There is some evidence that karyolymph
contributes to the formation of the spindle apparatus in plants.
Recently, Bernhard (1968) has described the localization of RNP (ribonucleoprotein) in the
nucleus, being mainly fibrillar and granular types. Besides, in nucleoplasm, there are (i)
interchromatic RNP fibrils located near the chromatin, (ii) interchromatic RNP granules
measuring about 250A in diameter and present around chromatin, (iii) Perichromatic RNP
granules of about 450A diameter, and (iv) large RNP coiled bodies and RNA granules of 300-
400A in diameter. In nuclei of Amoeba, there are present helices of about 700|i length and 300-
800A in diameter.
Nuclear constituents:
The nucleus contains RNA, DNA, proteins of two kinds, histone and nonhistone; some lipids;
various organic phosphorus compounds; and various inorganic compounds, mostly salts
(Davidson, 1976).
DNA:
In Salmon sperm heads, which are mostly occupied by nucleus, may constitute 48.5 % of the dry,
fat-free substance. Generally less than half the dry weight is DNA but the amount varies from
species to species. DNA remains constant and do not vary with nutrition or during starvation,
while proteins in nuclei vary, presistent (ultra-structure,)
RNAs:
Three kinds of RNA are present in cells: ribosomal (rRNA), transfer (tRNA) and messenger
(mRNA). RNA contains nucleotides of the purines, adenine and guanine and pyrimidines,
cytosine and uracil. In any of these RNAs the ratio of the bases to one another is constant for a
species. However, rRNA, tRNA and mRNA separated from one another disclose different base
ratio, indicating difference^ in their chemical composition.
The tRNAs have low molecular weight of about 25,000, the rRNAs have molecular weights
running to several million, and mRNAs are from half a million to several million. Differences
also exist in the affinities of the various RNAs for other molecules (Lehninger, 1975). The rRNA
is synthesized and assembled in the nucleolus, tRNAs and mRNAs are synthesized on the
chromosomes, and all the RNAs enter the cytoplasm through the nuclear pores.
Enzymes:
The nucleus contains a number of enzymes and performs metabolism, including synthesis of
DNA and various RNAs. Nucleus lacks the enzymes for aerobic metabolism that are found in
mitochondria, but it contains enzymes for anaerobic metabolism and for formation of high-
energy phosphates. It also contains enzymes for coenzyme synthesis (nicotinamide adenine
dinucleotide or NAD).
Proteins:
A variety of proteins is present in the nucleus: nucleoproteins, enzymes and structural proteins.
The nucleoproteins form two classes, deoxyribonucleoproteins and ribonucleoproteins.
Deoxyribonucleoproteins:
These largely form the chromosomes; consist primarily of histories and DNA in about equal
amounts. However, chromosomes also contain non-histone proteins in smaller amounts.
Unlike histones, most of the nonhistone proteins are acidic, and they vary qualitatively in
different cell types of the same organism.
Non-histone proteins are complexed to areas of DNA whose information is being expressed.
Hence it has been suggested that nonhistone proteins, along with chromosomal RNA which also
binds to certain active portions of DNA, may somehow be involved in the specific control of
gene expression. However, if nonhistone proteins do regulate gene expression, we do not know
how this occurs.
Nonhistone proteins contain the aromatic amino acid tryptophan. A considerable amount of the
contractile proteins actin, myosin, tropomyosin and tubulin are said to be present. Nonhistone
proteins have a more rapid turnover than histones do.
Both histones and nonhistone proteins are synthesized in the cytoplasm and enter the nucleus
through the nuclear envelope. Histones are synthesized only when DNA is replicated, whereas
nonhistone proteins are synthesized continuously. Histones induce a compact structure in the
chromosome.
Histones are also considered as stabilizers against heat damage (Tashiro and Kurakawa, 1976)
and against nucleases (Toczko et al., 1975). Activation and repression of genic expression are
thought to be carried out by nonhistone proteins. However, the mechanism by which this is done
in eukaryotic cells is less clear than it is in prokaryotic cells. All the proteins are synthesized in
the cytoplasm and then transported into the nucleus.
[III] Chromatin: DNA plus histones:
DNA is the main genetic constituent of cells, carrying information in a coded form from cell to
cell and from organism to organism. Within cells, DNA is not free but is complexed with
proteins in a structure called chromatin.
Chromatin:
It appears as a viscous, gelatinous substance which contains DNA, RNA, basic proteins called
histones, and nonhistone (more acidic) proteins. The content of RNA and non-histone protein is
variable between different chromatin preparations, but histone and DNA are always present in a
fixed ratio about one to one by weight. The nonhistone proteins are very heterogeneous; they
vary in different tissues and include RNA and DNA polymerase.
Histones:
These are small proteins which are basic because they have a high content (10 to 20 percent) of
the basic amino acids arginine and lysine. Being basic, histones bind tightly to DNA which is
acid. The four main histones, H2A, H2B and H4 are very similar in different species.
These four histones are present in equimolar amounts, two of each being present every 200 pairs
of DNA. Histone HI is not conserved between species and has tissue-specific forms. It is present
only once per 200 base pairs of DNA and is rather loosely associated with chromatin (it can be
eluted from DNA by adding low concentrations of salt).
It is not a component of the DNA-histone structural unit called the nucleosome core, but it is
bound to the linker segments of DNA that join neighbouring neucleosome. All histones consist
of a single polypeptide chain and have molecular weights between 11,000 and 21,000.
Functions of histones:
It is quite likely that they aid in the packing of DNA within the nucleus, and the binding of
histone to DNA serves to prevent the expression of hereditary information. All the hereditary
information present in any one cell is not expressed at all times. At any one moment, only a very
small proportion of the total information is expressed, i.e., only during a short period of
embryonic life. Prokaryotes do not have histones but a basic protein that might serve the same
function.
Chromatin has a repeating structure of beads about 10 nm in diameter connected by a string of
DNA. The 10 nm fibre represents the first level of organization of chromatin within cells.
The chromatin appears as thread-like, coiled and elongated structure which can be stained with
basic stains (e.g., basic fuchsin, orcein or Giemsa). These structures are termed chromatin fibres
or chromatin substance (Gr., chrome, colour). These are visible during interphase stage.
In interphase, chromatin of the chromosomes spreads out as a fine threads of linin, but at certain
regions, the chromatin remains condensed in the form of darkly stained chromatin mass. These
condensed regions are heterochromatic regions or heterochromatin, and the dispersed regions are
euchromatin. Both regions are formed of DNA.
During cell division, chromatin fibres become thick ribbon-like structure known as
chromosomes. The chromatin fibres remain twisted or form a network in the nucleoplasm. The
chromatin material is of two types:
1. Heterochromatin:
Certain regions of chromosomes containing chromatin mass become more darkly stained than
other regions. These particular darkly-stained parts are called as heterochromatic regions or
simply heterochromatin. The darkly staining regions show numerous bead-like bodies along the
chromosomes, called as chromomeres.
There are evidences that heterochromatic regions have a higher ribonucleic acid content than the
euchromatic regions. The heterochromatin is supposed to be metabolically and genetically inert
since it possesses small amount of DNA and large amount of RNA.
2. Euchromatin:
The light stained region of chromatin is called the euchromatic part or euchromatin. It contains
relatively more DNA.
[IV] Nucleolus:
Embedded in the matrix of nucleus there is a dense rounded, oval and acidophilic body called as
nucleolus, first described by Fontana in 1781 (nucleolus meaning small nucluns). Nucleolus
has no membrane of its own and is more dense than the surrounding nucleoplasm and hence is
distinctly visible.
Size:
The size of the nucleolus is related with the synthetic activity of the cell. Cells with little or no
synthetic activity (e.g., muscle cells, blastomeres, sperm cells, etc.) are found to contain smaller
or no nucleoli. On the other side, the secretory cells, neurons and oocytes which synthesize
proteins or other substances possess relatively large nucleoli. In the living cell, nucleoli are
highly refringent bodies due to large concentration of solid material.
Number:
The number of nucleoli in nucleus depends upon the number of chromosomes and species. There
may be only one nucleolus in many plants and animal cells for each haploid set of chromosomes.
In others, there may be two or more nucleoli for each haploid set of chromosomes. In man, there
are two pairs of nucleoli in each diploid nucleus.
The nucleolus stains with pyronine and other stains and absorbs ultraviolet light at 260 nm.
Treatment with ribonuclease shows that the capacity to absorb this basophilic stain and
ultravoilet radiation depends on the presence of RNA.
Position:
There are certain heterochromatic regions of specific chromosomes found in association with the
nucleolus, constituting nucleolar organizing regions of chromosomes. This indicates that
although all of the chromosomes contribute to the formation of nucleolar material but these
certain organizing regions are responsible for its constitution. The nucleolar organizers are now
known to contain the genes coding for 18S, 28S and 5.8S and RNAs.
The 18S, 5.8S and 28S RNAs are synthesized in the nucleolus, where as 5S RNA is synthesized
on the chromosomes outside the nucleolus and the 70 ribosomal proteins are synthesized in the
cytoplasm.
All these components migrate to the nucleolus, where they are assembled into ribosomes and
transported to the cytoplasm. The nucleolar organizer is usually located in a secondary
constriction on the chromosome, i. e., in a chromosomal site that becomes less condensed during
mitosis.
There may be more than one nucleolar organizer region among the chromatin strands in a
nucleus, and hence more than one nucleolus may be present in a single cell.
The nucleolus is known to be the cellular site for the synthesis of ribosomal RNA, the RNA
component of the ribosomes.
1. Chemical composition of nucleolus:
Maggio, Palade (1963) and Vincent (1952) have described the chemical composition of
nucleolus. In liver cells, nucleoli contain RNA as 3-5%, whereas in pea embryo, RNA contents
are 10% or 20% of total nuclear RNA. The protein components include mainly phosphoproteins.
Histones have not been reported in the isolated nucleoli but Tandler (1962) has described a high
concentration of orthophosphates, which may serve as a precursor of the RNA phosphorus.
Besides, Sirlin, Jacob and Tandler (1963) have verified the presence of acid phosphatase,
nucleoside, phosphorylase, DPN synthetase and RNA methylase enzymes, although some of
them may be lacking in different nucleoli.
2. Types of nucleoli:
According to E.B. Wilson, there are two or three categories of nucleoli, namely plasmosomes or
true nucleoli and karyosomes (Ogata) or chromatin-nucleoli, etc.
(a) Plasmosomes:
The plasmosomes are of oxyphilic nature, i.e., get staind with acidic stains. Their outer part is
transparent, called as halo, and inner one is dense.
(b) Karyosomes (Ogata):
These are basophilic in nature, i.e., get stained with basic dyes. Montgomery calls them as
chromatin nucleoli. They are associated with the chromosomes formation during cell division.
The karyosomes are of 3 kinds: Firstly, they may be net-knot type (i.e., in the form of nodes
composing spireme threads).
Secondly, they include chromosome nucleoli which occur only in gametocytes and are
spheroidal bodies. They represent a group of chromosomes in a condensed form during resting
phase. The third category includes karyospheres which are called nucleolusnoyanx by Carnoy.
They are also spheroidal bodies having basic chromatin.
(c) Amphinucleoli:
Sometimes plasmosomes and karyosomes are closely associated to form a double nucleolus or
amphinucleolus. These are very common in the eggs of molluscs, annelids, etc.
3. Components of nucleolus:
During cell division the nucleolus generally disappears during the first stage or prophase stage,
but it reappears in the daughter cells. Morphologically, two components may be defined in a
nucleolus:
(a) Pars amorpha:
This is a component of nucleolus which first disappears but reappears at the end of division. This
is the amorphous part which begins to disappear just prior to the breakdown of nuclear
membrane during cell division and reappears in daughter nuclei as division completes.
(b) Nucleolonema:
This is the second and permanent component which does not disappear but remains persistent
throughout the cell cycle. It is a filamentous structure having 80A fibrils with which 150A
particles of ribonucleoprotein are attached.

4. Ultra-structure of nucleolus:
Recently, as a result of high resolution studies (ultra-structure), it has been demonstrated that
there exists a definite submicroscopic organization within the nucleolus. The various elements as
described by De Robertis et al., (1971) are as follows:
(a) Granular portion being made up of dense granules arranged peripherally and measuring about
150-200 A in diameter. This granular zone occupies the peripheral region of the nucleolus, which
is surrounded by the associated chromatin.
It consists of ribonucleic proteins. Granular zone contains ribosome precursor particles at various
stages of assembly and processing. Once the ribosomal subunits are mature, they are released
from the nucleolus and exit into the cytoplasm through the nuclear pores.
(b) Fibrillar portion having the fibrils of 50-80A length and composed of ribonucleoprotein. This
region is also known as nucleolonema. It generally occupies the central region of the nucleolus.
Both the granular and fibrillar zones are digested by ribonuclease. Fibrillar zone contains rRNA
genes that uncoil from the chromosome and penetrate the nucleolus, together with nascent RNA
molecules attached to it.
(c) Amorphous region having low electron density (dielectronic) and formed of certain proteins
which can be hydrolysed easily by the pepsin enzyme.
(d) Nucleolus associated chromatin present around the nucleolus which sometimes extends into
the nucleolus. The nucleolus has no limiting membrane and the calcium ions are believed to
maintain the intact organization of the nucleolus.
The nucleolus also contains vacuoles having no limiting membrane and it possesses certain
proteins. The chromatin of the nucleolar organizer becomes associated with fibrillar and granular
material to form the nucleolus.
During cellular reproduction, when the chromatin is condensed into discrete chromosomes,
nucleoli usually disappear, but they re-form later in the interphase nuclei of the new daughter
cells.
5. Functions of nucleolus:
It serves the following functions:
(a) Role in mitosis:
The nucleolus plays a significant part in mitosis. In grasshopper neuroblasts, there are two
nucleoli in each nucleus. If any one of these nucleoli is injured by ultra-violet radiation or by
other source, the mitosis is permanently stopped. It gives a clear evidence that both nucleoli must
be present for the initiation of mitosis.
(b) Help in protein synthesis:
The nucleoli help in protein synthesis by the formation of ribonucleic acid. This ribonucleic acid
(RNA) plays an important role in the formation of proteins. The cells with a high rate of protein
synthesis have large nucleoli with a high RNA content, whereas the cells with a low rate of
protein synthesis have small undeveloped nucleoli.
The nucleolus is a factory for ribosomes. The nucleolus is formed at the nucleolar organizer,
which is a chromosomal site that contains tandem (one behind another) repeats of the genes
coding for 18S and 28S rRNA.
This DNA becomes uncoiled and penetrates the nucleolus, where it is actively transcribed. In
addition, other ribosomal components such as 5S RNA and the ribosomal proteins, which are
synthesized in other parts of the cell, converge on the nucleolus, where the assembly of
ribosomal subunits starts.
(c) As intermediator in the transmission of genetic information:
The nucleolus serves as an intermediary for carrying the genetic information from generation to
generation. Evidences for this function are given by the structure of the salivary gland cell of
Bradysia mycorum (Sciaridae).
These gland cells contain large multiple nucleoli which arise from the primary nucleoli
associated with the chromosomes. These primary nucleoli become detached from chromosomes
and become congregated to form multiple nucleoli which are considered to be the main site of
genetic information.
6. Biogenesis of nucleolus cycle:
During mitosis the nucleoli undergo cyclic changes. Nucleoli are formed around the DNA loop
that extends from the nucleolar organizer. Thus, nucleoli are tormed from loops of one or more
chromosomes combining with specific proteins. There may be several nucleoli per cell, but
frequently they tend to fuse into one or a few nucleoli at this stage.
During late prophase the DNA loop containing rRNA genes gradually retracts and coils into the
nucleolar organizer of the corresponding chromosome. Since this DNA is highly extended as a
result of intense RNA synthesis, the nucleolar organizer region is one of the last to undergo
condensation, thus producing a secondary constriction on the chromosome.
The fibrillar and granular regions are gradually dispersed into the nucleoplasm. After the cell
divides, during telophase, the nucleolar organizer DNA uncoils and the nucleolus is reassembled.
The chromatin material of the nucleolar organizer carries the information that directs the
formation of the nucleolus and of the ribosomal RNA. In the nucleolus, the newly synthesized
ribosomal RNA (probably the fibrillar material of nucleolus) combines with proteins, which are
apparently synthesized in the cytoplasm and transported to the nucleolus, to form particles
(nucleolar granules) that are precursors to cytoplasmic ribosomes.
[V] Endosomes:
These are rather smaller chromatin bodies present in the nucleoplasm of nucleus. They are like
nucleolus but smaller in size, showing changeable structure.
Cytoplasmic Filaments: Composition, Distribution and Functions
By Anandita Biology
Cytoplasmic filaments are elongate, un-branched, proteinaceous strands that consist of bundles
or groups of protein molecules sometimes wound into a helical shape. Although organized arrays
of cytoplasmic filaments were first described for muscle cells and have been most extensively
studied in this tissue, it is now apparent that essentially all eukaryotic cells contain these
filaments.
Cytoplasmic filaments can be subdivided into three major classes based on filament size;
these are:
(1) Microfilaments (also called thin filaments), which have a diameter of about 6 n and are
composed primarily of the globular protein actin;
(2) Intermediate filaments, which have diameters between 7 and 11 nm and, depending on the
tissue source, are formed from five different classes of proteins (see below); and
(3) Myosin filaments (also called thick filaments), which have diameters up to 22 nm and are
rich in the fibrous protein myosin.
Muscle cells, which are especially differentiated for contraction, are rich in microfilaments and
myosin filaments, but all cells contain cytoplasmic filaments and it has been shown that actin can
account for as much as 20% of the total cell protein in some non-muscle cells.
Microfilaments:
Although actin molecules are globular proteins (i.e., G-actin), in microfilaments the actin
molecules are arranged to form two interwoven helical chains, each of which is called a proto-
filament (Fig. 23-6); in this polymeric form, the protein is called F-actin. Microfilaments are
intimately associated with all cellular activities that involve movement.

This is vividly demonstrated when cells are treated with cytochalasin B. In the presence of this
substance, microfilaments dissociate and the loss of the microfilaments is accompanied by a loss
of certain cellular functions. Some of the more common processes sensitive to cytochalasin B are
phagocytosis, pinocytosis, and exocytosis, cytokinesis, cytoplasmic streaming (in plant cells),
movements of microvilli, cilia, and flagella, movements of the cytoskeleton, and of course
muscle contraction.
Intermediate Filaments:
Five different classes of proteins comprise the intermediate filaments of cells and tissues; these
are desmin in muscle cells (one type of protein of MW 52,000), vimentin in mesenchymal cells
(one protein of MW 53,000), cytokeratins in epithelial cells (more than a dozen different proteins
having MWs ranging from 40,000 to 70,000), neuro-filament proteins in nerve cells (three
proteins having MWs of 65,000, 105,000, and 135,000), and glial fibrillary acidic protein in
astrocytes (one protein of MW 50,000).
Although a dynamic role cannot be excluded for the intermediate filaments, in most cells they
appear to play a structural role. For example, in epidermal and mesenchymal cells, bundles of
intermediate filaments form a basketlike weave that encircles the nucleus and is intimately
associated with the nuclear envelope at the nuclear pores.
From there the filaments appear to radiate to the margins of the cells, where they terminate at
desmosomes. Because of their intimate association with the nuclear pores, a role in mediating the
transport of materials between the nucleus and the cyotplasm cannot be precluded.
In nerve cells, the long axes of the intermediate filaments are oriented parallel to the long axes of
the cells long processes (e.g., axons and dendrites) and are believed to provide these processes
with a structural framework.
Myosin (Thick) Filaments:
Striated, smooth, and cardiac muscle cells contain vast numbers of cytoplasmic filaments that
function during the contraction of these cells. Most of these filaments are microfilaments (i.e.,
thin filaments formed primarily from F-actin but also. containing the proteins tropomyosin and
troponin) and thick filaments (formed from myosin).
The two types of cytoplasmic filaments are arranged in parallel rows and interact with each other
through cross-bridges that enable the filaments to slide past one another and effect a shortening
(i.e., contraction) of the cell. The contraction of striated muscle cells shortens the muscle, which
then moves the skeleton. It is in striated muscle cells that the number of filaments is greatest and
their geometric distribution most highly ordered.
As seen in Figure 23-7, equally spaced about each thick filament are six thin filaments. Units of
several hundred thick and thin filaments are bundled together to form myofibrils, and each cell
contains many hundreds of myofibrils.

Cytokinesis:
Cell division in animal cells involves two separate mechanisms mitosis and cytoplasmic
cleavage, or cytokinesis. In animal cells, cytokinesis begins toward the end of anaphase and is
characterized by the appearance of a constriction about the midline of the spindle. The
constriction deepens as the plasma membrane moves inward at the cleavage furrow. Material
collecting at the midline of the spindle becomes quite dense, forming a structure known as the
mid-body (Fig. 23-8). Just before the in-folding edges of the plasma membrane meet and fuse,
the mid-body disappears.

The furrowing or pinching-in of the plasma membrane at the cleavage furrow is reminiscent of
the action of a purse string or of a rubber band tightening about a soft object. The process clearly
involves the action of microfilaments and these are seen in abundance in the area of the cleavage
furrow. Microfilament involvement is also supported by the observation that cytochalasin B
inhibits the process.
The presence of a band of microfilaments, the contractile ring, just underneath the plasma
membrane in the area of constriction can be seen in the electron photomicrographs of Figure 23-
9. Here Arbacia sea urchin eggs are observed at various stages of cleavage. Prior to the onset of
cleavage, no microfilaments are seen in the area that is about to constrict (Figs. 23-9a and 23-
9d).

However, once cleavage begins, microfilaments quickly appear about the area of constriction,
forming the contractile ring (compare Figs. 23-9d and 23-9e). Because the bundles of
microfilaments are arranged in a circle just below the cleavage furrow, their long axes run
perpendicular to the plane of the sections seen in Figures 23-9b and 23-9e.
As cleavage is completed, the spindle fibers and mid-body fade and the contractile ring and
microfilaments disappear (Fig. 23-9c). The molecular basis of the constriction that characterizes
cytokinesis in animal cells remains uncertain, but the clear presence of actin and the suspected
presence of myosin has led to the speculation that the mechanism involves filament sliding
similar to that which occurs during muscle contraction.
Plasma Membrane Movement:
Intestinal epithelial cells have thousands of small fingerlike projections called microvilli that
cyclically shorten and extend into the lumen of the intestine. This action facilitates the absorption
of digested food by greatly increasing the surface area of the cells.
Microfilaments in the intestinal microvilli are bundled together to form a core by the interaction
of F-actin with the two proteins villin and fimbrin. The microfilament bundle runs parallel to the
length of the microvillus and is anchored to the plasma membrane at the microvillus apical tip.
No myosin appears to be present in the microvilli. At the base of the microvilli is another
microfilament network called the terminal web. Myosin has been localized in the terminal web
and its interaction there with actin is believed to support the microvilli and perhaps also create a
contractile mechanism that lowers the microvilli into the cell. A second possibility is that in the
presence of high Ca
2+
, villin can act to cleave F-actin into short fragments. This would cause the
microvilli to collapse into the cell in a mechanism that is independent of myosin. Calmodulin has
also been implicated in the regulation of movements of microvilli.
Amoeboid Movement:
The locomotion of amoebae, slime molds, white blood cells, and a number of other cells involves
the formation of pseudopodialarge cytoplasmic extensions from the main body of the cell and
into which the remaining cytoplasm subsequently streams. Most amoeboid cells from more than
one pseudopod at a time, but continued locomotion in one direction require the reversal of the
process in the non-dominant pseudopodia.
In pseudopodia-forming cells, the outer margin of the cytoplasm, called ectoplasm, is very
viscous or gel-like and is generally free of granules and other cytoplasmic inclusions; the
remaining cytoplasm, called endoplasm, is more fluid or sol-like. During locomotion, the fluid
endoplasm flows forward into the advancing pseudopod, and as it reaches the anterior end of the
pseudopod, it flows laterally and is converted into gel, thereby forming new ectoplasm. At the
rear of the moving cell, in a region called the uroid, the ectoplasm solates, streams deeper into
the cell, and becomes endoplasm (Fig. 23-10).

Two different views have dominated attempts to explain amoeboid movement. According to the
front- zone contraction theory of R. D. Allen and others, the endoplasm in the anterior region
of a pseudopod undergoes contraction as it is transformed into ectoplasm. In effect, this
contraction pulls the endoplasm forward and into the pseudopod. An older view advanced in the
1920s by S.O. Mast and called the ectoplasmic tube contraction theory suggests that the
ectoplasm toward the rear of the cell contracts, pushing the endoplasm forward.
Regardless of the site of origin of the force that causes the cytoplasm to stream forward,
overwhelming evidence implicates actin microfilaments in the process. For example, the actin-
binding drug cytochalasin B inhibits amoeboid movement.
The presence of myosin filaments in amoeboid cells has also been reported as has an ATPase
similar to one known to be important in the chemistry of muscle contraction. Contractions of the
ectoplasm of amoeboid cells can be induced by adding ATP and Ca
2 +
to the cells. Although not
fully understood, amoeboid movement appears to use an actinmyosin contractile system similar
to that of muscle cells.

An amoeboid like but much slower form of locomotion is also exhibited by mammalian
connective tissue cells (i.e., fibroblasts) grown in culture. As a fibroblast moves across a surface
(e.g., the culture dish), the cell continuously forms sheetlike extensions, called lamellipodia, and
micro spikes at its advancing edge (Figs. 23-11 and 23-12). Some of the lamellipodia attach to
the surface (at anchor points called adhesion plaques) and others sweep backward over the cells
upper surface in a wavelike motion called surface ruffling.

Electron-microscopic studies reveal that the lamellipodia contain large numbers of actin
microfilaments, and like true amoeboid motion, the movements of fibroblasts are inhibited by
cytochalasin B. As the leading edge of the cell stretches forward, the trailing edge is drawn out
into long retraction fibers that are eventually pulled free from their points of attachment to the
underlying surface. The latter action may leave behind small pieces of plasma membrane and
cytoplasm.
Capping, Membrane Flow, and Cell Locomotion:
Substances bound to mobile plasma membrane receptors migrate laterally within the membrane
and are concentrated in clathrin-coated pits prior to endocytosis. During endocytosis, these
receptors and the ligands that they have bound are internalized along with small pieces of the
plasma membrane. The receptors are eventually returned to the plasma membrane during
exocytosis, which also restores membrane surface area lost during endocytosis. In non-motile
cells, endocytosis and exocytosis occur at positions scattered over much of the cell surface.
Many membrane proteins avoid internalization during endocytosis because their rates of
diffusion within the membrane preclude their concentration within the coated pits. These
membrane proteins are referred to antibodies against non-cycling membrane proteins, the
proteins are cross-linked to form large aggregates. Because the diffusion rates of cross-linked
proteins are so much lower than the diffusion rates of individual proteins, the aggregate can be
clustered together.
In motile cells, cross-linked proteins cluster at one point on the cell surface called a cap, and the
process is called capping. Capping is a general phenomenon displayed by aggregates in the
plasma membrane and is not peculiar to certain proteins; non-motile cells, however, do not cap
cross-linked surface proteins. Cytochalasin B and another drug called colchicine prevent
capping, suggesting that membrane flow involves the actions of microfilaments (and perhaps
also microtubules).
When fibroblasts are allowed to migrate over a substrate littered with tiny carbon particles, the
particles become attached to the upper surface of extended lamellipodia and are slowly swept
backward toward the trailing edge of the cell. This implies that the membrane as a whole is
flowing backward. It has therefore been suggested that a moving fibroblast internalizes portions
of the plasma membrane-at the rear of the cell and reinserts this membrane at the leading edge of
the cell (Fig. 23-13).

This would provide a mechanism by which the cell could move itself forward along a substrate,
for if the cell forms transient connections with the substrate at its anterior end, the forces that
cause the membrane to flow backward would simultaneously propel the cell forward. The action
would be similar to that of a tractor or tank track