Journal of Neuroendocrinology 20 (Suppl.

1), 90–93
ª 2008 The Authors. Journal Compilation ª 2008 Blackwell Publishing Ltd
REVIEW ARTICLE

The Cannabinoid System and Male Reproductive Functions

M. Rossato, C. Pagano and R. Vettor
Endocrine-Metabolic Laboratory, Clinica Medica 3, Department of Medical and Surgical Sciences, University of Padova, Padova, Italy.

Journal of
Neuroendocrinology
Correspondence to:
Dr Marco Rossato, Endocrine-
Metabolic Laboratory, Clinica Medica
3, Department of Medical and
Surgical Sciences, University of
Padova, Via Ospedale 105, 35128
Padova, Italy (e-mail: marco.rossato@
unipd.it).
Cannabinoids, the main active components of marijuana, have been shown to exert different
adverse effects on male reproduction both in vertebrates and invertebrates. In vivo, cannabinoids
exert negative effects on hypothalamic–hypophyseal reproductive hormone secretion and testi-
cular endocrine and exocrine functions. Furthermore, a large amount of experimental data
obtained in vitro have clearly shown that cannabinoids negatively influence important sperm
functions, including motility and acrosome reaction, two fundamental processes necessary for
oocyte fertilisation. These inhibitory effects are mediated by the direct action of cannabinoids on
sperm through the activation of the cannabinoid receptor subtype CNR1 that has been shown
to be expressed in mature sperm. In the present paper, we briefly review the effects of cannabi-
noids and endocannabinoids, a particular group of endogenously produced cannabinoids, on
male reproductive function.
Key words: cannabinoids, sperm, male reproduction.
doi: 10.1111/j.1365-2826.2008.01680.x

Marijuana, the most widely used recreational drug worldwide (1),
contains a large number of different active compounds known as
cannabinoids, and D9-tetrahydocannabinol (THC) has been recog-
nised as the main active molecule (2). Beside natural cannabi-
noids, different tissues can synthesise a group of related
unsaturated fatty acid derivatives named endocannabinoids that
act as endogenous ligands for the cannabinoid receptors. This so
called ‘endocannabinoid system’ regulates many different func-
tions in all tissues via interaction with two specific receptors,
cannabinoid receptor subtype 1 (CNR1) and cannabinoid receptor
subtype 2 (CNR2), whose tissue distribution and function have
been previously described (3–5). A number of different endocann-
abinoids have been isolated within biological fluids, anandamide
(N-arachidonoylethanolamine, AEA) and 2-arachidonoyl glycerol
(2-AG) being the main components (3). In particular, AEA is the
most abundant in reproductive secretions, thus suggesting a pos-
sible role for the endocannabinoid system in reproduction (6).
CNR receptors have been identified along male reproductive tract:
CNR1 receptors have been detected in the testis, prostate and
vas deferens (7–10). Furthermore, functional CNR1 receptors have
been found in sperm in different vertebrate and invertebrate spe-
cies (10–12). To date, unequivocal data have been reported show-
ing that cannabinoids control male reproductive functions both
directly and indirectly, depending of the expression of the CNR
receptor subtypes along the tissues controlling the testicular
function (Table 1) with important final effects on fertility in the
male (10–12).
Cannabinoids and spermatogenesis
Cannabinoids and hypothalamus-pituitary-testis axis
Spermatogenesis is a long a complex process controlled by a num-
ber of different mechanisms (13). Gonadotrophins (FSH [follicle
stimulating hormone] and LH [luteinising hormone]) play a crucial
role in the induction and maintenance of spermatogenesis in mam-
mals including man (14), involving a stimulatory effect of Sertoli
and Leydig cells and a complex series of secreting events that fol-
low. It is known that in males, cannabis smoking decreases plasma
LH levels, leading to decreased plasma testosterone (15), although
contradictory results have been reported (16, and references
therein). FSH secretion has also been shown to be modulated by
cannabinoids (17) and the combined effects on the gonadotrophins
lead to decreased sperm count in males that has been consistently
observed in men who smoke cannabis (18).
The effects of cannabinoids on gonadotrophin secretion seem to
be localised upstream to the pituitary, and, in particular, at the pre-
optic area of the hypothalamus where CNR1 receptors have been
localised and where LH releasing hormone (LHRH)-secreting
neurones reside (19). At this level, cannabinoids seem to modulate
 Cannabinoids and male reproduction 91

Table 1. Expression of the Cannabinoid Receptor Subtypes in the Tissues grating brain and testicular messages are modulated by cannabi-
Participating in the Control of the Mammalian Male Reproductive Tract noids at both the hypothalamic–pituitary and the testicular level.
Functions.

Cannabinoid receptor Cannabinoids and sperm functions

Tissue subtype Reference
Cannabinoids and sperm motility
Hypothalamus
GnRH secreting cells CNR1 ⁄ CNR2 (20) Since the first report published by Perez et al. (26) in 1981, experi-
Pituitary mental data have accumulated confirming that cannabinoids nega-
Gonadotrophs CNR1 (21) tively influence the motility of sperm in different mammalian
Testis species. Apart from observations obtained in sea urchins showing
Germ cells CNR1 (10) no significant effects of cannabinoids on sperm motility (12), these
Sertoli cells CNR2 (24) substances inhibit sperm motility in different mammalian species,
Leydig cells CNR1 (8) including man (10, 27). The observations on the negative effects of
Epididymis n.d. –
cannabinoids on sperm motility were reported in the mid 1970s
Vas deferens CNR1 (38)
and early 1980s; however, CNR1 and CNR2 were not identified until
Seminal vesicles n.d. –
Prostate CNR1 (37)
the early 1990s, when expression of CNR1 was demonstrated in the
Corpus cavernosum CNR1 (39) testis (28). Only recently, taking a pharmacological experimental
approach, the inhibitory effects of cannabinoids on mammalian
CNR1, cannabinoid receptor subtype 1; CNR2, cannabinoid receptor subtype sperm motility have been attributed to CNR1 activation (10). Very
2; n.d., not determined. recently, Cobellis et al. (11) replicated these results in sperm from
Rana esculenta, taking the same pharmacological approach.
negatively the activity of LHRH-secreting neurones by direct and Furthermore, the same authors extended these observations by
indirect mechanisms (20). Beyond the well known effect of canna- means of a molecular genetic experimental design, showing that
binoids on gonadotrophin secretion at the hypoyhalamic level mod- sperm from CNR1-knockout mice show a dramatic increase in
ulating gonadal function, previous observations have localised motility percentages in the caput epididymis. This clearly demon-
CNR1 in the anterior pituitary, suggesting a direct effect of canna- strates that the lack of CNR1 signalling increases the acquisition of
binoids on LH and FSH secretion at the pituitary level (21). sperm motility (9).
Although the studies evaluating the effects of cannabinoids on
sperm motility are not abundant, it seems that in general the acti-
Cannabinoids and male germ cells
vation of CNR1 inhibits motility in invertebrate and vertebrate
Evidence primarily from animal studies has shown that CNR1 is sperm, at least in vitro. Contradictory results have been reported,
expressed in germ cells, from spermatogonia to mature sperm. For however, regarding the effects of cannabinoids ingestion, as well as
instance Gye et al. (8) demonstrated the presence of this receptor marijuana smoking, on human sperm motility in vivo. Variation in
in spermatogonia, primary spermatocytes and sperm in mouse tes- these results may be owing to the experimental design or the small
tis, and these observations have been replicated by Cobellis et al. number of subjects participating to the different clinical studies
(11) in the testis of invertebrates. Current knowledge clearly shows (29).
that cannabinoids negatively influence the spermatogenetic process,
reducing germ cell proliferation and reproductive organ weight (11,
Cannabinoids and sperm capacitation
22). Sertoli cells constitute one of the main components of the
seminiferous tubule, and their number – fixed at puberty – is clo- After ejaculation, sperm have to undergo a complex – and still
sely related to the total sperm production. Sertoli cells surround unclear – series of biochemical and morphological events collec-
and nurse germinal cells during their maturation. By mediating the tively known as ‘capacitation’, before they are capable of fertilising
actions of all hormonal stimuli regulating the spermatogenetic pro- an egg (30). This includes remodelling the lipid plasma membrane,
cess they provide a finely tuned microenvironment leading to germ which is required to improve the ability of sperm to respond to
cell maturation from spermatogonium to mature sperm (23). stimuli that induce the acrosome reaction, facilitating the interac-
CNR2 has been recently described in Sertoli cells, modulating tion with the egg. These changes are activated by still not fully elu-
apoptosis in these cells (24). Moreover, Leydig cells have been cidated mechanisms involving the adenylate cyclase ⁄ cAMP ⁄ protein
shown to express CNR1, which, when activated, induces a reduction kinase A (PKA) signalling pathway, leading to tyrosine phosphoryla-
of testosterone production, thus further inhibiting the spermatoge- tion of different proteins (31). These processes are inhibited by
netic process (because high local levels of testosterone are required cannabinoids and CNR1 activation (27, 31). These effects are
to maintain spermatogenesis (8)). Furthermore, testosterone is specific: they are counteracted by sperm pre-incubation with the
necessary for epididymis and seminal vesicle activity, which CNR1 antagonist SR141716.
influence important sperm functions such as motility (25). Thus, the
complex events coordinating the male spermatogenesis and inte-

ª 2008 The Authors. Journal Compilation ª 2008 Blackwell Publishing Ltd, Journal of Neuroendocrinology, 20 (Suppl. 1), 90–93
92 M. Rossato et al.
Physiologically, sperm capacitation occurs within the female gen-
ital tract and it has been demonstrate that sperm leaving seminal
plasma and reaching the uterus and oviduct swim in an external
milieu presenting progressively reduced concentrations of endo-
cannabinoids (6). As sperm approach the egg, they encounter the
lowest concentrations of endocannabinoids present in the female
genital tract, thus suggesting that the activation of the endo-
cannabinoid system in sperm seems to maintain the sperm in an
uncapacitated, quiescent condition before interacting with the egg
(10).
Cannabinoids and sperm acrosome reaction
Sperm capacitation is a fundamental pre-requisite for exocytosis of
the acrosome, a specialised secretory granule located at the apex of
sperm head (32). Sperm acrosome reaction occurs as the sperm
faces the egg, and leads to the release of proteolytic enzymes nec-
essary for penetration of the egg’s coats, the exposure of specia-
lised sperm membrane sites that allow attachment and fusion with
the egg’s plasma membrane and then to fertilisation (33). Sperm
that do not undergo the acrosome reaction are not able to bind to
the egg’s zona pellucida and cannot fertilise it. Substantially homo-
geneous data exist indicating that CNR1 activation reduce the abil-
ity of sperm to undergo acrosome reaction both in vertebrates and
invertebrates (10, 12, 27, 31). These data further confirm the inhibi-
tory effects of cannabinoids on sperm fertilising ability.
Importantly, since cannabinoids do not affect fertility of eggs,
these effects are directed only to sperm (27). The mechanisms
involved in cannabinoid inhibition of the sperm acrosome reaction
are still unknown. It is well known that ion channels and regulation
of ion fluxes through the sperm plasma membrane are crucial for
the acrosome reaction, and the most important physiological regu-
lator appears to be Ca2+ (34). In this regard we have previously
shown that the endocannabinoid AEA did not modify intracellular
Ca2+ concentrations in human sperm, possibly ruling out any inter-
fering effect of cannabinoids on Ca2+ signalling (10). CNR1 is a
G-protein-coupled receptor that has been shown to inhibit adenyl-
ate cyclase activity (3). Intracellular levels of cAMP rise during acro-
some reaction, cell-permeant cAMP analogues induce the acrosome
reaction, and pharmacological inhibitors of cAMP-dependent PKA
reduce this exocytotic event (35, 36): these observations suggest
that the adenylate cyclase ⁄ cAMP ⁄ PKA system plays a primary role
in the signalling pathway leading to sperm acrosome reaction and
egg fertilisation. The inhibitory effect of cannabinoids on this fun-
damental signalling pathway, as recently described in mammalian
sperm (31), might explain the negative effect these agents have,
through CNR1 activation, on sperm capacitating and acrosome
reaction.
Conclusions
Marijuana is the most commonly used recreational drug worldwide
and it is now well recognised that the active components of
marijuana – as well as endogenously synthesised endocannabinoids
– negatively affect sperm functions, both in invertebrates and ver-
ª 2008 The Authors. Journal Compilation ª 2008 Blackwell Publishing Ltd, Journal of Neuroendocrinology, 20 (Suppl. 1), 90–93
tebrates, impairing the fertilisation process and thus reproductive
function. This is important, given the prevalence of marijuana
smoking particularly in the young. These observations, together
with the demonstration that the male reproductive tract possesses
the whole enzymatic machinery for the synthesis and metabolism
of endocannabinoids, could suggest the existence of particular clini-
cal situations, beyond marijuana smoking, characterised by a patho-
logical increase of endocannabinoids within the male reproductive
tract, that could lead to an impairment of sperm function.
Furthermore, if we consider that the use of cannabinoids has
been legally authorised in some countries for the treatment of
some pathological conditions (such as appetite stimulation in
patients with AIDS, treatment of nausea and vomiting, reduction of
symptoms in some neurological diseases, reducing intraocular pres-
sure in glaucoma patients, treatment of depression and pain), the
possible negative effects of these therapeutic cannabinoid agonists
on human male fertility have to be taken into careful account.
Conflicts of interest
MR, CP and RV have received speaker’s fees from Sanofi-Aventis.
Received: 7 January 2008,
accepted 8 February 2008
References
1 Di Marzo V. A brief history of cannabinoid and endocannabinoid phar-
macology as inspired by the work of British scientists. Trends Pharmacol
Sci 2006; 27: 134–140.
2 Di Marzo V, De Petrocellis L, Bisogno T. The biosynthesis, fate and phar-
macological properties of endocannabinoids. Handb Exp Pharmacol
2005; 168: 147–185.
3 Felder CC, Glass M. Cannabinoid receptors and their endogenous agon-
ists. Annu Rev Pharmacol Toxicol 1998; 38: 179–200.
4 Pertwee RG. Pharmacology of cannabinoid CB1 and CB2 receptors. Phar-
macol Ther 1997; 74: 129–180.
5 McPartland JM, Glass M, Pertwee RG. Meta-analysis of cannabinoid
ligand binding affinity and receptor distribution: interspecies differences.
Br J Pharmacol 2007; 152: 583–593.
6 Schuel H, Burkman LJ, Lippes J, Crickard K, Forester E, Piomelli D, Giuffri-
da A. N-Acylethanolamines in human reproductive fluids. Chem Phys
Lipids 2002; 121: 211–227.
7 Pertwee RG, Ross RA, Craib SJ, Thomas A. Cannabidiol antagonizes can-
nabinoid receptor agonists and noradrenaline in the mouse vas deferens.
Eur J Pharmacol 2002; 456: 99–106.
8 Gye MC, Kang HH, Kang HJ. Expression of cannabinoid receptor 1 in
mouse testes. Arch Androl 2005; 51: 247–255.
9 Ricci G, Cacciola G, Altucci L, Meccariello R, Pierantoni R, Fasano S,
Cobellis G. Endocannabinoid control of sperm motility: the role of epi-
didymus. Gen Comp Endocrinol 2007; 153: 320–322.
10 Rossato M, Ion Popa F, Ferigo M, Clari G, Foresta C. Human sperm
express cannabinoid receptor Cb1, the activation of which inhibits motil-
ity, acrosome reaction, and mitochondrial function. J Clin Endocrinol
Metab 2005; 90: 984–991.
11 Cobellis G, Cacciola G, Scarpa D, Meccariello R, Chianese R, Franzoni MF,
Mackie K, Pierantoni R, Fasano S. Endocannabinoid system in frog and
rodent testis: type-1 cannabinoid receptor and fatty acid amide hydro-
lase activity in male germ cells. Biol Reprod 2006; 75: 82–89.

12 Schuel H, Goldstein E, Mechoulam R, Zimmerman AM, Zimmerman S.
Anandamide (arachidonylethanolamide), a brain cannabinoid receptor
agonist, reduces sperm fertilizing capacity in sea urchins by inhibiting
the acrosome reaction. Proc Natl Acad Sci U S A 1994; 91: 7678–7682.
13 Foresta C, Bettella A, Merico M, Garolla A, Plebani M, Ferlin A, Rossato
M. FSH in the treatment of oligozoospermia. Mol Cell Endocrinol 2000;
161: 89–97.
14 Foresta C, Bettella A, Rossato M, La Sala G, De Paoli M, Plebani M. Inhi-
bin B plasma concentrations in oligozoospermic subjects before and
after therapy with follicle stimulating hormone. Hum Reprod 1999; 14:
906–912.
15 Kolodny RC, Masters WH, Kolodner RM, Toro G. Depression of plasma
testosterone levels after chronic intensive marihuana use. N Engl J Med
1974; 290: 872–874.
16 Park B, McPartland JM, Glass M. Cannabis, cannabinoids and reproduc-
tion. Prostaglandins Leukot Essent Fatty Acids 2004; 70: 189–197.
17 Wenger T, Rettori V, Snyder GD, Dalterio S, McCann SM. Effects of D-9-
tetrahydrocannabinol on the hypothalamic-pituitary control of luteiniz-
ing hormone and follicle-stimulating hormone secretion in adult male
rats. Neuroendocrinology 1987; 46: 488–493.
18 Hembree WC III, Nahas GG, Zeidenberg P, Huang HF. Changes in human
spermatozoa associated with high dose marihuana smoking. Adv Biosci
1978; 23: 429–439.
19 Matsuda LA, Bonner TI, Lolait SJ. Localization of cannabinoid receptor
mRNA in rat brain. J Comp Neurol 1993; 327: 535–550.
20 Gammon CM, Freeman GM Jr, Xie W, Petersen SL, Wetsel WC. Regula-
tion of gonadotropin-releasing hormone secretion by cannabinoids.
Endocrinology 2005; 146: 4491–4499.
21 Wenger T, Fernández-Ruiz JJ, Ramos JA. Immunocytochemical demon-
stration of CB1 cannabinoid receptors in the anterior lobe of the pitui-
tary gland. J Neuroendocrinol 1999; 11: 873–878.
22 Maccarrone M, Falciglia K, Di Rienzo M, Finazzi-Agrò A. Endocannabi-
noids, hormone-cytokine networks and human fertility. Prostaglandins
Leukot Essent Fatty Acids 2002; 66: 309–317.
23 Brinster RL. Male germline stem cells: from mice to men. Science 2007;
316: 404–405.
24 Maccarrone M, Cecconi S, Rossi G, Battista N, Pauselli R, Finazzi-Agrò A.
Anandamide activity and degradation are regulated by early postnatal
aging and follicle-stimulating hormone in mouse Sertoli cells. Endocri-
nology 2003; 144: 20–28.
25 Higgins SJ, Burchell JM. Effects of testosterone on messenger ribonucleic
acid and protein synthesis in rat seminal vesicle. Biochem J 1978; 174:
543–551.
ª 2008 The Authors. Journal Compilation ª 2008 Blackwell Publishing Ltd, Journal of Neuroendocrinology, 20 (Suppl. 1), 90–93
Cannabinoids and male reproduction 93
26 Perez LE, Smith CG, Asch RH. delta 9-tetrahydrocannabinol inhibits fruc-
tose utilization and motility in human,rhesus monkey, and rabbit sperm
in vitro. Fertil Steril 1981; 35: 703–705.
27 Schuel H, Burkman LJ, Lippes J, Crickard K, Mahony MC, Giuffrida A,
Picone RP, Makriyannis A. Evidence that anandamide-signaling regulates
human sperm functions required for fertilization. Mol Reprod Dev 2002;
63: 376–387.
28 Gérard CM, Mollereau C, Vassart G, Parmentier M. Molecular cloning of
a human cannabinoid receptor which is also expressed in testis. Biochem
J 1991; 279: 129–134.
29 Whan LB, West MC, McClure N, Lewis SE. Effects of delta-9 tetrahydro-
cannabinol, the primary psychoactive cannabinoid in marijuana, on
human sperm function in vitro. Fertil Steril 2006; 85: 653–660.
30 Salicioni AM, Platt MD, Wertheimer EV, Arcelay E, Allaire A, Sosnik J, Vis-
conti PE. Signalling pathways involved in sperm capacitation. Soc Reprod
Fertil Suppl 2007; 65: 245–259.
31 Maccarrone M, Barboni B, Paradisi A, Bernabò N, Gasperi V, Pistilli MG,
Fezza F, Lucidi P, Mattioli M. Characterization of the endocannabinoid
system in boar spermatozoa and implications for sperm capacitation and
acrosome reaction. J Cell Sci 2005; 118: 4393–4404.
32 Yanagimachi R. Mammalian fertilization. In: Knobil E, Neill JD, eds. The
Physiology of Reproduction. New York: Raven Press, 1994: 189–281.
33 Wassarman PM, Jovine L, Qi H, Williams Z, Darie C, Litscher ES. Recent
aspects of mammalian fertilization research. Mol Cell Endocrinol 2005;
234: 95–103.
34 Darszon A, Labarca P, Nishigaki T, Espinosa F. Ion channels in sperm
physiology. Physiol Rev 1999; 79: 481–510.
35 Beltrán C, Vacquier VD, Moy G, Chen Y, Buck J, Levin LR, Darszon A. Par-
ticulate and soluble adenylyl cyclases participate in the sperm acrosome
reaction. Biochem Biophys Res Commun 2007; 358: 1128–1135.
36 De Jonge CJ, Han HL, Lawrie H, Mack SR, Zaneveld LJ. Modulation of the
human sperm acrosome reaction by effectors of the adenylate cyclase ⁄
cyclic AMP second-messenger pathway. J Exp Zool 1991; 258: 113–125.
37 Tokanovic S, Malone DT, Ventura S. Stimulation of epithelial CB1 recep-
tors inhibits contractions of the rat prostate gland. Br J Pharmacol
2007; 150: 227–234.
38 Christopoulos A, Coles P, Lay L, Lew MJ, Angus JA. Pharmacological
analysis of cannabinoid receptor activity in the rat vas deferens. Br J
Pharmacol 2001; 132: 1281–1291.
39 Ghasemi M, Sadeghipour H, Dehpour AR. Anandamide improves the
impaired nitric oxide-mediated neurogenic relaxation of the corpus cav-
ernosum in diabetic rats: involvement of cannabinoid CB1 and vanilloid
VR1 receptors. BJU Int 2007; 100: 1385–1390.