Reviews of Reproduction (1999) 4, 4855

Leptin and reproduction

Iain J. Clarke and B. A. Henry
Prince Henrys Institute of Medical Research, PO Box 5152,Clayton, Victoria 3168, Australia

In the few years since leptin was identied as a satiety factor in rodents, it has been implicated in the regulation of various physiological processes. Leptin has been shown to promote sexual maturation in rodent species and a role in reproduction has been investigated at various sites within the hypothalamopituitarygonadal axis. This review considers the evidence that leptin (or alteration in amount of body fat) can affect reproduction. There is evidence that leptin plays a permissive role in the onset of puberty, probably through action on the hypothalamus, where leptin receptors are found in cells that express appetite-regulating peptides. There is little evidence that leptin has a positive effect on the pituitary gonadotrophs and the gonads. There is also very little indication that leptin acts in an acute manner to regulate reproduction in the short term. It seems more likely that leptin is a barometer of body condition that sends signals to the brain. Studies in vitro have shown negative effects on ovarian steroid production and there are no reports of effects on testicular function. Leptin concentrations in plasma increase in women during pregnancy, owing to production by the placenta but the functional signicance of this is unknown. A number of factors that affect the production and action of leptin have yet to be studied in detail.

Leptin is a peptide produced by adipocytes and achieves hormonal status by virtue of its secretion into the bloodstream (Considine et al., 1996). It is a potent satiety factor (Campeld et al., 1995; Halaas et al., 1995) but may serve other functions, such as the regulation of neuroendocrine systems and the gonads. This brief review will outline the sites within the hypothalamopituitary axis at which leptin may inuence reproductive function, indicating the location of leptin receptors and detailing effects within the hypothalamopituitarygonadal axis (Fig. 1). Since leptin may affect the secretion of various hormones, the way it inuences reproduction could either be direct or indirect (Fig. 1). For example, leptin could regulate the secretion of growth hormone (GH) or prolactin (see below), which could have indirect effects on the gonad (Adashi, 1995; Linzer and Arey, 1995).

Bodyweight, nutritional status and reproductive function

The concentrations of leptin in plasma have been shown to be a direct reection of the amount of body fat (Dagago-Jack et al., 1996; McGregor et al., 1996), and reproductive function is substantially affected by bodyweight and nutritional status (Martin et al., 1994; Yen, in press). Gonadotrophin secretion is reduced in animals and humans that are nutritionally restricted (Cameron and Nobisch, 1991; Loucks and Heath, 1994; Foster et al., 1995), and gonadal function is compromised. However, ovarian and testicular function can be enhanced in sheep by the feeding of high protein supplements (Martin et al., 1994; Pomares et al., 1995). Nevertheless, the relationship between bodyweight and reproductive function is not straightforward. One well cited study is that of the Kung San (Bushmen) of Africa in whom there is a seasonal cycle of fertility that

relates to the availability of food (Van der Walt et al., 1978). This may not be an example of a simple function between nutrition and reproduction because, at the time when the women of this population are anovulatory, they not only have nutritional restriction but also are subjected to strenuous physical exercise while seeking food. Furthermore, there are situations in which reduced function of the reproductive axis under chronic undernutrition can eventually be resolved. In one example, we (Barker-Gibb and Clarke, 1996) have shown that the plasma LH and FSH concentrations in fat and thin sheep were not signicantly different after maintenance on either ad libitum or restricted diets for 16 months. Furthermore, studies of women on the Indian sub-continent have shown that fertility is reduced in cases of chronic undernutrition, but that the women eventually become fertile without any improvement in their diet (Shetty, 1946). This is a salient issue because it points to the likelihood that long-term maintenance of signicant differences in bodyfat (and, presumably, plasma leptin concentrations) will not always indicate different fertility rates. Studies on the hypothalamopituitary axis have not identied any single blood-borne component that could act as a metabolic indicator of the reproductive axis at this level (Schreihofer et al., 1996). The concentrations of sugar in the blood need to be maintained within physiological limits if GnRH secretion is to be normal (Clarke et al., 1990), but this cannot be regarded as a specic metabolic signal because reduced blood glucose concentrations will cause generalized gluconeuropaenia. It is possible that leptin provides an appropriate signal of metabolic status to the reproductive system, causing altered function with altered bodyweight (fatness) or nutritional status. However, there is no convincing evidence that leptin signals are an acute regulator

1999 Journals of Reproduction and Fertility 1359-6004/99 $15.00

Leptin and reproduction

49

of reproductive hormone secretion. In addition, the debate continues as to whether leptin is a primary stimulus of the reproductive axis or acts as a permissive factor (especially with respect to puberty).

Indirect

Direct

Effects of leptin on the hypothalamus and pituitary gland

The leptin receptor has various splice variants, with the long form being functional in terms of intracellular signalling (Lee et al., 1996) and the short form possibly important for transport within the choroid plexus (Lee et al., 1996; Malik and Young, 1996). In the hypothalamus, the long form of the receptor is found in the arcuate nucleus and has been localized to neurones containing neuropeptide Y (NPY) (Mercer et al., 1996) and pro-opiomelanocortin (POMC) (Cheung et al., 1997a). It is also found in other parts of the hypothalamus in dened cell types (Mercer et al., 1996; Dyer et al., 1997; Hakansson et al., 1998) and in the visceral nuclei of the brainstem (Goldstone et al., 1997), so action is not conned to the arcuate nucleus. The exact percentage of each cell type that contains the leptin receptor has not been ascertained and it is unclear with which parts of the brain these leptin receptor-containing cells communicate. Cells from the arcuate nucleus project to a number of brain regions, including the lateral hypothalamic nuclei, the paraventricular nucleus and the pre-optic area. Therefore, cells of the arcuate nucleus may regulate the function of cells in the appetite regulating areas (for example, the ventromedial hypothalamus), stress centres (for example, the paraventricular nucleus) and areas important for reproduction (for example, the pre-optic area). With respect to reproduction, the question of whether leptin has a direct effect on GnRH-containing cells of the brain is important. In most species, most GnRH cell bodies are found in the preoptic area, but also extend rostrally into the diagonal band of Broca, ventrally into the organum vasculosum of the lamina terminalis and caudally into the medial septum. A smaller number of cells is found in the mediobasal hypothalamus of most species, with the notable exception of primates. Zamorano et al. (1997) have reported that mRNA encoding the leptin receptor may be found in immortalized GnRH cells by reverse transcriptionpolymerase chain reaction (RTPCR) analysis but it is not known whether the receptor is expressed in normal GnRH cells. Leptin can inuence the secretion of GnRH from the immortalized cells provided they are rst transfected with the long form of the leptin receptor (S. Moenter, personal communication). The lateral hypothalamus and the visceral nuclei of the rostral brain stem are brain areas important for the regulation of appetite and food intake (Morley, 1987). Within these regions, large numbers of hypothalamic peptides and neurotransmitters are involved in the regulation of appetite, and the integration of these various factors is obviously complex (for review, see Morley, 1987). The possible means by which leptin acts to regulate appetite as well as its neuroendocrine function are indicated (Fig. 2). Many of the peptides and neurotransmitters implicated in the regulation of appetite and food intake are also known to inuence the secretion of GnRH (for review, see Morley, 1987; Kalra, 1993) and there may well be common mechanisms that subserve both functions. A good example of

Releasing and inhibiting factors GnRH GH PRL GH PRL

Appetiteregulating peptides

Brain

Pituitary

L L Fat

L L

L Gonads

Fig. 1. Leptin (L) may act at different levels and affect the reproductive system directly. It may also act to regulate the secretion of other hormones such as growth hormone (GH) or prolactin (PRL), which then act on the gonads (indirect action).

commonality is found in the case of NPY. This peptide is the most potent orexigenic peptide known, and its expression in the arcuate nucleus is reduced by leptin (Ahima et al., 1996), consistent with the role of leptin as a satiety factor. NPY cells contain oestrogen receptors and project to GnRH cell bodies (Kalra, 1993). NPY also inhibits the secretion of LH when an intracerebroventricular (i.c.v.) infusion is given to gonadectomized rats (Kalra, 1993) and sheep (Barker-Gibb et al., 1995). A stimulatory effect of NPY is seen in steroid-primed ovariectomized rats and there is increased secretion of NPY into the hypophysial portal blood at the time of the preovulatory LH surge (for review, see Kalra, 1993). An i.c.v. infusion of NPY antiserum blocks the preovulatory LH surge in sheep (Porter et al., 1993). In the broadest sense, a role for NPY can be invoked in the generation of the preovulatory surge, and a decit or reduction of this peptide would compromise the event. If leptin plays a permissive or stimulatory role in the regulation of GnRH secretion, the reduction of NPY expression by leptin is not consistent with this. However, it is possible that leptin regulates the secretion of GnRH through a number of other neuronal systems, such as melanin-concentrating hormone (MCH) (Gonzalez et al., 1997) and POMC (Sahu, 1998 and Thornton et al., 1997). POMC produces the endogenous opioid, -endorphin which is a negative regulator of GnRH secretion (Kalra, 1993). However, opioids stimulate appetite (Morley, 1987). If leptin inhibits the expression of POMC, this would be consistent with effects that stimulate the reproductive axis while inhibiting appetite.

50

I. J. Clarke and B. A. Henry

Hypophysiotrophic peptides
GnRH CRF AVP TRH GRF SRIF

? NPY

Appetiteregulating peptides
EOP MCH GLP-1 GAL

Pituitary hormones
LH FSH GH TSH PRL ACTH/end

Fat

Testes/ovaries

Fig. 2. Leptin (L) may regulate appetite and reproduction in a co-ordinated manner, acting on cells that express neuropeptide Y (NPY), endogenous opioid peptides (EOP), melanin-concentrating hormone (MCH), glucagon-like peptide 1 (GLP-1) or galanin (GAL), and these neuropeptides could, in turn, act on hypophysiotrophic systems within the hypothalamus. ACTH/end, adrenocorticotrophic hormone/-endorphin; AVP, arginine vasopressin; CRF, corticotrophin-releasing factor; GH, growth hormone; GnRH, gonadotrophin-releasing hormone; GRF, growth hormone-releasing factor: PRL, prolactin; SRIF, somatostatin; TRH, thyroid-releasing hormone; TSH, thyroid-stimulating hormone.

Various peptides regulating appetite localize to the lateral hypothalamus and may also regulate GnRH secretion. A good example is MCH, which is found in this region (Bittencourt et al., 1992). MCH was originally shown to inhibit food intake (Presse et al., 1996) but was later found to stimulate appetite (Qu et al., 1996; Rossi et al., 1997). There is a greater amount of mRNA encoding MCH in the lateral hypothalamus of ob/ob animals (Qu et al., 1996). With regard to a role in reproduction, Gonzalez et al. (1997) presented evidence that MCH stimulates GnRH secretion after central administration to rats. Yu et al. (1997) reported an effect of leptin on the secretion of GnRH from arcuate nucleusmedian eminence fragments in vitro. A very small (< 20%) stimulatory effect of leptin was seen with lower doses (10121010mol l1), whereas higher doses had no effect. With i.c.v. injection of leptin at one dose (10 g), a stimulatory effect on the maximum increase in plasma LH during the rst hour was seen in ovariectomized female rats treated with 10 g oestrogen 72 h earlier. Whether these animals were in a state of negative or of positive feedback is not clear and there did not appear to be any consideration of diurnal effects on LH secretion in this model. The reduction in pulsatile LH secretion that occurs with fasting in

ovariectomized rats can be overcome by treatment with leptin (Nagatani et al., in press), suggesting that leptin may convey information to centres within the brain that regulate GnRH secretion. Furthermore, these authors showed that the leptin effect could be obtained whether or not the animals were treated with oestrogen, suggesting that leptin does not necessarily modulate the feedback effect of this gonadal steroid. Carro et al. (1997) have shown that i.c.v. infusion of leptin antiserum decreases LH pulsatility in female rats and suppresses ovarian cyclic activity. Taken together, these data suggest that leptin plays a role in the regulation of GnRHLH secretion in rats. In normally fed ovariectomized ewes, a 3 day i.c.v. infusion of leptin had no effect on the secretion of gonadotrophins, prolactin, GH or cortisol, despite a highly signicant reduction in food intake that drove the animals into negative energy balance (Henry et al., 1998). These data suggest that the effects of high doses of leptin on the centres in the brain that regulate appetite need not necessarily result in effects on the secretion of hypophysiotrophic hormones. Further work needs to be done in non-rodent species before a generalized role for leptin in the regulation of reproduction can be invoked. The long form of the leptin receptor is expressed in the pituitary gland (Dyer et al., 1997; Zamorano et al., 1997) but the the cell types in which it is localized have not yet been identied. Roh et al. (in press) have shown that leptin stimulates basal GH secretion but inhibits growth hormone-releasing factor-stimulated GH secretion from ovine pituitary somatotrophs in culture, providing some indication that this cell type may express the receptor. Yu et al. (1997) have presented evidence that leptin can inuence gonadotrophin secretion from acutely prepared rat hemi-pituitaries. It would be interesting to know whether similar results could be obtained with cells that were conditioned in culture, or in animal preparations in which the pituitary gland is isolated from the brain and receives physiological pulsatile GnRH stimulation. It is most likely that any physiological response to leptin will be the outcome of a variety of actions at different levels. A simplistic representation of this concept is shown (Fig. 3). One way to reduce the number of variables in the overall equation is to identify all the cell types that contain leptin receptors and then study the various parameters in these cells. It should be remembered that acute excursions in the plasma concentrations of leptin are not seen, for example, after a meal. There are diurnal changes but it seems more likely that concentrations that are maintained over days or weeks will be the most relevant in terms of providing a setting in which a physiological system may work. Therefore, it will be important to study the effects of leptin administered over extended periods rather than observing the effects of a single injection. In this way, the role of leptin will become more clearly elucidated and may be related to the situation that pertains in obesity or extreme reduction in body mass index.

Effects of leptin on the gonads

The short form of the leptin receptor is found in both the ovary (Karlsson et al., 1997) and the testis (Hoggard et al., 1997). Leptin appears to inhibit ovarian function on the basis of a variety of parameters. For example, it inhibits the insulininduced secretion of progesterone and androstenedione from

Leptin and reproduction

51

Brain HPTH
(e.g. GnRH)

ARP II
(e.g. MCH)

ARP I
(e.g. NPY)

Pituitary gland L L (e.g. gonadotroph) LH/FSH L Fat Gonads L

Steroids

(e.g. granulosa cell)

Fig. 3. Leptin (L) may act at various levels. Within the brain, it may regulate appetite regulating peptides (ARP) and there could be effects in series (for example, ARPIARPII). Any of these systems could affect the secretion of hypophysiotrophic hormones (HPTH) such as GnRH. Leptin receptors (red bars) have been identied in the brain, within the pituitary gland and in the gonads. The cell types in which the receptors are found within the pituitary gland has not yet been determined. Any of the actions of leptin may be modied by steroids, but such effects are not yet well dened. MCH, melanin-concentrating hormone; NPY, neuropeptide Y.

cultured bovine thecal cells (Spicer and Francisco, 1998) and the insulin-induced production of oestradiol and progesterone from bovine granulosa cells (Spicer and Francisco, 1997). Zachow and Magofn (1997) showed an inhibitory effect of leptin on combined insulin-like growth factor I and FSHstimulated oestrogen production from rat granulosa cells. In cultured human granulosa cells, Karlsson et al. (1997) found that leptin inhibited LH-stimulated oestradiol secretion, leading them to suggest that the short form of the receptor acts in a dominant negative manner in the ovary. We are not aware of any published results of effects of leptin on the testis. It remains to be determined whether leptin has a positive or a negative effect on gonadal function in vivo. However, on the basis of available information, it seems that a direct effect of leptin on the gonads cannot explain the advancement of puberty or the restoration of reproductive function in the ob/ob mouse (see below).

Advancement of puberty by leptin treatment

Various studies have demonstrated the importance of leptin in the initiation of puberty in rodents. The majority of genetically

obese (ob/ob) male mice and all ob/ob female mice are infertile, probably because of reduced hypothalamic GnRH content and low plasma gonadotrophin concentrations. This infertility can be corrected in both sexes by the administration of leptin (Chehab et al., 1996; Mounzih et al., 1997), which stimulates the secretion of gonadotrophins, and gonadal function (Barash et al., 1996). Treatment with recombinant leptin also advances puberty in normal female mice (Chehab et al., 1997) and rats (Cheung et al., 1997b) and allows the onset of puberty in severely foodrestricted rats (Gruaz et al., 1998). It has been suggested that leptin is the signal that informs the brain that metabolic stores are adequate for the initiation of reproductive function (Chehab et al., 1997), but there are other possibilities. First, the aforementioned studies do not rule out a direct effect of leptin on the gonads (see above). In spite of indications from other studies that leptin has a negative effect on steroidogenesis, Zamorano et al. (1997) reported that the treatment of ob/ob mice with leptin increases the amount of mRNA for side-chain cleavage enzyme and 17-hydroxylase, so there may be a stimulatory effect in vivo. Second, leptin may be permissive and not necessarily an initiator of puberty (Cheung

52

I. J. Clarke and B. A. Henry

et al., 1997b). In support of this hypothesis, plasma leptin concentrations in monkeys do not change around the onset of puberty (Plant and Durrant, 1997). Studies of plasma leptin concentrations in humans across the prepubertal years and into early puberty also suggest that leptin may play a facilitatory role in the initiation of puberty (Clayton et al., 1997; Matkovic et al., 1997; Ricardo et al., 1997), but whether it acts at the brain, the pituitary gland or the gonads is not delineated. A strong case can be made for leptin as a permissive factor in the initiation of puberty.

the human menstrual cycle, reaching peak values during the luteal phase (Hardie et al., 1997). A profound increase in plasma leptin concentrations is seen during pregnancy (Hardie et al., 1997) and this appears to be due to the production of leptin by the placenta (Masuzaki et al., 1997) and may be related to the well-being of the fetus. There does not appear to be any placental production of leptin in mice, even though plasma concentrations increase during pregnancy due to increased production by adipose tissue (Kawai et al., 1997; Tomimatsu et al., 1997).

Relationship of leptin to plasma sex steroid concentrations and acute responses of the reproductive axis to altered nutritional status and leptin production
The feeding of high protein foodstuff to animals causes an acute response in terms of gonadotrophin release, and the use of ushing to improve ovulation rates has long been practised in agriculture. Whether these effects are involved in the secretion of leptin is not clear. There are various examples of an effect of short-term nutritional manipulation on reproductive function. Cameron and colleagues have shown that gonadotrophin secretion is severely compromised in monkeys that miss a meal, but restoration of function occurs immediately upon re-feeding (Cameron and Nobisch, 1991). This acute effect of feeding may be related to leptin production but this remains to be determined. The fasted animals have lowered blood insulin and blood glucose concentrations that are restored with re-feeding, and these could lead to a short-term increase in plasma leptin concentrations. Intragastric infusion of casein and lipids, which does not restore blood glucose concentrations but does restore insulin concentrations, is also able to re-initiate pulsatile LH secretion (Schreihofer et al., 1996). The stimulation of LH secretion on re-feeding may be due to recognition of the metabolic state of the animal but no particular blood borne factor has been identied. It seems unlikely that this factor is leptin because plasma concentrations in fasted individuals do not increase for a number of hours after re-feeding (Weigle et al., 1997) and the LH response in monkeys occurs earlier than this. Martin et al. (1994) have shown that the feeding of lupin grain, which has high digestible amounts of energy and protein, is able to stimulate gonadotrophin secretion in sheep over a period of 5 days. This is a slower response than is seen in re-fed monkeys and may well be due to stimulation of the GnRH system by leptin. The relationship between feeding and insulin and leptin production is directly pertinent to the above issues. Insulin stimulates the secretion of leptin by adipocytes but this response is slow. The increase in insulin secretion after feeding does not stimulate leptin production in the short term (Dagogo-Jack et al., 1996) but this occurs over a number of hours. The gradual increase in plasma leptin concentrations that occurs throughout the day and into the evening is consistent with this. Likewise, the nadir that is reached by early morning is consistent with the lack of food intake during the night. Plasma concentrations of leptin are lower in human males than in females (Ricardo et al., 1997), perhaps due to the multifarious effects of testosterone in males (Arslanian and Suprasongsin, 1997). Leptin concentrations may vary across

Important points to consider Effects on appetite versus effects of leptin

Since reduced food intake will compromise reproduction and leptin reduces food intake, it is important to delineate these two effects. If leptin acts as a messenger of metabolic status and stimulates the onset of puberty, how is this reconciled with its appetite-reducing properties? It is quite possible that some of its functions are achieved at concentrations below those required for appetite reduction. Careful studies need to be conducted in which the effects of leptin are examined at doses below those that reduce food intake.

Season
There is a distinct seasonal appetite cycle in most species (Forbes, 1982). Studies need to be performed that investigate whether the effects of leptin are different at different times of the year and whether there are seasonal alterations in the expression of appetite-regulating peptides within the hypothalamus.

Effects of steroids
Steroids are known to inuence the expression of peptides within the hypothalamus that affect both reproduction and appetite. For example, expression of NPY is increased by the treatment of castrated male rats with testosterone (Sahu et al., 1992) and decreased by the treatment of ovariectomized female rats with oestrogen (Baskin et al., 1995). Since NPY is thought to be central to the regulation of appetite, the steroid status of the animal will be important with respect to the effects of leptin either on appetite or reproduction. Indeed, Bennett et al. (1998) suggest that oestrogen can inuence the response to leptin by altering the ratio of the short and long forms of the receptor in the central nervous system. Insulin-induced hypoglycaemia reduces LH secretion in intact or ovariectomized, oestrogen-treated monkeys but has no effect in ovariectomized monkeys (Chen et al., 1992). Whether this relates to an altered expression of appetite-regulating peptides in the brain with different steroidal background is not known.

Differences between and within species

Bronson (1985) has pointed out that rodents carry very little body fat and that a short period of fasting can have a profound effect on their metabolism that leads to a very rapid

Leptin and reproduction

53

shut-down of their reproductive function. However, human females have ideally 22% bodyfat (for review, see Yen, in press) and fasting may not have such severe short-term consequences. Ruminants, such as sheep, are able to experience substantial undernutrition and yet maintain plasma glucose and insulin concentrations and not compromise gonadotrophin secretion. Harris et al. (1998) reported that ob/ob mice are more sensitive to leptin than are lean mice for a wide range of parameters. These are important points to consider when the inuence of bodyweight and metabolic status on the reproductive axis are examined, and extrapolation of results in rodent models to humans may not be valid. Even within species there are signicant factors to consider. Adam et al. (1998) examined the effect of undernutrition on the onset of puberty in a primitive breed of sheep (Soay) compared with an improved breed. Restricted nutrition inuenced the onset of puberty in the improved breed but not in the Soay, leading to speculation that leptin plays a more substantial role in the initiation of puberty in the improved breed.

Conclusions
Since leptin was identied as a satiety factor, a substantial volume of literature has appeared on its role in the regulation of appetite and various other functions. On the basis of receptor localization and cellular responses, there is good evidence that leptin acts within the hypothalamus. Although it is an attractive notion that leptin interacts with the neuroendocrine system to signal metabolic status to the hypothalamic and pituitary cells that regulate reproduction, much work needs to be done to substantiate this hypothesis. Well performed work in vivo needs to be carried out to determine the effects on GnRH secretion in various physiological settings, and the possibility of a direct effect on pituitary gonadotrophs needs to be carefully considered. Effects on the gonads are possible but, once again, further work in vivo is required to examine this in more detail. To date, most studies have indicated negative effects on gonadal function. A wide range of factors (for example, species, season, metabolic status and steroidal status) need to be considered in the conduct of such experiments. There is very little indication that leptin acts in an acute manner to regulate reproduction in the short term and it seems more likely that leptin is a barometer of body condition, signalling to the brain. In this way, leptin may be a permissive factor with respect to the onset of puberty.

References
Key references are identied by asterisks. Adam CL, Findlay PA, Kyle CE and Young P (1998) Effect of restricted nutrition on timing of puberty in female Soay sheep Journal of Reproduction and Fertility 112 3137 Adashi EY (1995) Ovary as a site of growth hormone reception and action. In The Somatotrophic Axis and the Reproductive Process in Health and Disease pp 175182 Eds EY Adashi and MO Thorner. Serono Symposia USA. Springer Verlag, New York *Ahima RS, Prabakaran D, Mantzoros C, Qu D, Lowell B, Maratos-Flier E and Flier JS (1996) Role of leptin in the neuroendocrine response to fasting Nature (London) 382 250252

Arslanian S and Suprasongsin C (1997) Testosterone treatment in adolescents with delayed puberty: changes in body composition, protein, fat, and glucose metabolism Journal of Clinical Endocrinology and Metabolism 82 32133220 Barash IA, Cheung CC, Weigle DS, Ren H, Kabigting EB, Kuijper JL, Clifton DK, Steiner RA (1996) Leptin is a metabolic signal to the reproductive system Endocrinology 137 31443147 Barker-Gibb ML and Clarke IJ (1996) Increased galanin and neuropeptide Y immunoreactivity within the hypothalamus of ovariectomized ewes following a prolonged period of reduced body weight correlates with changes in plasma growth hormone levels but not gonadotropin levels Neuroendocrinology 64 194207 Barker-Gibb ML, Scott CJ, Boublik JH and Clarke IJ (1995) The role of neuropeptide Y (NPY) in the control of LH secretion in the ewe with respect to season, NPY receptor subtype and the site of action in the hypothalamus Journal of Endocrinology 147 565579 Baskin DG, Norwood BJ, Schwartz MW and Koerker DJ (1995) Estradiol inhibits the increase of hypothalamic neuropeptide Y messenger ribonucleic acid expression induced by weight loss in ovariectomized rats Endocrinology 136 55475554 Bennett PA, Lindell K, Karlsson C, Robinson ICAF, Carlsson LMS and Carlsson B (1998) Differential expression and regulation of leptin receptor isoforms in the rat brain: effects of fasting and oestrogen Neuroendocrinology 67 2936 Bittencourt JC, Presse F, Arias C, Peto C, Vaughn J, Nahon JL, Vale W, Sawchenko PE (1992) The melanin-concentrating hormone system of the rat brain: an immuno- and hybridisation histochemical characterisation Journal of Comparative Neurology 319 218245 Bronson FH (1985) Mammalian reproduction: an ecological perspective Biology of Reproduction 32 126 Cameron JL and Nobisch C (1991) Suppression of pulsatile luteinizing hormone and testosterone secretion during short term food restriction in the adult male rhesus monkey (Macaca mulatta) Endocrinology 128 15321540 Campeld LA, Smith FJ, Guisez Y, Devos R and Burn P (1995) Recombinant mouse OB protein: evidence for a peripheral signal linking adiposity and central neural networks Science 269 546549 Carro E, Pinilla L, Seoane LM, Considine RV, Aguilar E, Casanueva FF and Dieguez C (1997) Inuence of endogenous leptin tone on the estrous cycle and luteinizing hormone pulsatility in female rats Neuroendocrinology 66 375377 Chehab F, Lim ME and Lu R (1996) Correction of the sterility defect in homozygous obese female mice by treatment with the human recombinant leptin. Nature Genetics 12 318320 Chehab FF, Mounzih K, Lu R and Lim ME (1997) Early onset of reproductive function in normal female mice treated with leptin Science 275 8890 Chen M-D, OByrne KT, Chiappini J and Knobil E (1992) Hypoglycemic stress and gonadotropin-releasing hormone pulse generator activity in the rhesus monkey: role of the ovary Neuroendocrinology 56 666673 Cheung CC, Clifton DK, Steiner RA (1997a) Proopiomelanocortin neurons are direct targets for leptin in the hypothalamus Endocrinology 138 44894492 *Cheung CC, Thornton JE, Kuijper JL, Weigle DS, Clifton DK, Steiner RA (1997b) Leptin is a metabolic gate for the onset of puberty in the female rat Endocrinology 138 855858 Clarke IJ, Horton RJE and Doughton BW (1990) An investigation of the mechanism by which insulin-induced hypoglycemia decreases LH secretion in ovariectomized ewes Endocrinology 127 14701476 Clayton PE, Gill MS, Hall CM, Tillmann V, Whatmore AJ and Price DA (1997) Serum leptin throughout childhood and adolescence Clinical Endocrinology 46 727733 Considine RV, Sinha MK, Heiman ML et al. (1996) The protein product of Ob gene is elevated in human obesity: effect of weight loss fasting and feeding New England Journal of Medicine 334 292295 Dagogo-Jack S, Fanelli C, Paramore D, Brothers J and Landt M (1996) Plasma leptin and insulin relationships in obese and non-obese humans Diabetes 45 695698 Dyer CJ, Simmons JM, Matteri RL and Keisler DH (1997) Leptin receptor mRNA is expressed in ewe anterior pituitary and adipose tissues and is differentially expressed in hypothalamic regions of well-fed and feedrestricted ewes Domestic Animal Endocrinology 14 119128 Forbes JM (1982) Effects of lighting patterns on growth, lactation and food intake of sheep, cattle and deer Livestock Production Science 9 361374

54

I. J. Clarke and B. A. Henry

Matkovic V, Ilich JZ, Skugor M, Badenhop NE, Goel P, Clairmont A, Klisovic D, Nahhas and Landoll JD (1997) Leptin is inversely related to age at menarche in human females Journal of Clinical Endocrinology and Metabolism 82 32393245 Mercer JG, Hoggard N, Williams LM, Lawrence CB, Hannah LT, Morgan PJ and Trayhurn P (1996) Coexpression of leptin receptor and preproneuropeptide Y mRNA in arcuate nucleus of mouse hypothalamus Journal of Neuroendocrinology 8 733735 *Morley JE (1987) Neuropeptide regulation of appetite and weight Endocrine Reviews 8 256277 Mounzih K Lu R and Chehab FF (1997) Leptin treatment rescues the sterility of genetically obese ob/ob males Endocrinology 138 11901193. *Nagatani S, Guthikonda P, Thompson RC, Tsukamura H, Maeda K-I and Foster DL (1998) Evidence for GnRH regulation by leptin: leptin administration prevents reduced pulsatile LH secretion during fasting Neuroendocrinology 67 370376 Plant TM and Durrant AR (1997) Circulating leptin does not appear to provide a signal for triggering the initiation of puberty in the male rhesus monkey Endocrinology 138 45054508 Pomares CC, Galloway DB, Holmes JHG, Clarke IJ and Tilbrook AJ (1995) Lupin and cowpea supplements for growth, wool production and reproduction in rams Australian Journal of Experimental Biology 35 447452 Porter DWF, Naylor AM, McNeilly AS and Lincoln DW (1993) Endocrine actions of central neuropeptide Y in the ewe: activation of the hypothalamopituitaryadrenal axis by exogenous neuropeptide Y and role of endogenous neuropeptide Y in the secretion of luteinizing hormone during the oestrous cycle Journal of Neuroendocrinology 5 163174 Presse F, Sorokovosky I, Max J-P, Nicolaidis S and Nahon J-L (1996) Melanin-concentrating hormone is a potent anorectic peptide regulated by food-deprivation and glucopeinia in the rat Neuroscience 71 735745 Qu D, Ludwig DS, Gammeltoft S, Piper M, Pelleymounter MA, Cullen MJ, Mathes WF, Przypek J, Kanarek R and Maratos-Flier E (1996) A role for melanin-concentration hormone in the central regulation of feeding behaviour Nature (London) 380 243247 Ricardo V, Garcia-Mayor M, Andrade A, Rios M, Lage M, Dieguez C and Casanueva FF (1997) Serum leptin levels in normal children: relationship to age, gender, body mass index, pituitarygonadal hormones, and pubertal stage Journal of Clinical Endocrinology and Metabolism 82 28492855 Roh S-G, Clarke IJ, Goding JW and Chen C The effect of leptin in growth hormone secretion in primary cultured ovine pituitary cells Neuroendocrinology (in press) Rossi M, Choi SJ, OShea D, Miyoshi T, Ghatei MA and Bloom SR (1997) Melanin-concentration hormone acutely stimulates feeding, but chronic administration has no effect on body weight Endocrinology 138 351355 Sahu A (1998) Evidence suggesting that galanin (GAL), melanin-concentrating hormone (MCH), neurotensin (NT), proopiomelanocortin (POMC) and neuropeptide Y (NPY) are targets of leptin signalling in the hypothalamus Endocrinology 139 795798 Sahu A, Phelps CP, White JD, Crowley WR, Kalra SP and Kalra PS (1992) Steroidal regulation of hypothalamic neuropeptide Y release and gene expression Endocrinology 130 33313336 Schreihofer, DA, Renda, F and Cameron, JL (1996) Feeding-induced stimulation of luteinizing hormone secretion in male rhesus monkeys is not dependent on a rise in blood glucose concentration Endocrinology 137 37703776 Shetty PS (1946) Adaptive changes in basal metabolic rate and lean body mass in chronic undernutrition. Human starvation and its consequences Journal of the American Diet Association 22 582587 Spicer LJ and Francisco CC (1997) The adipose obese gene product, leptin: evidence of a direct inhibitory role in ovarian function Endocrinology 138 33743379 Spicer LJ and Francisco CC (1998) Adipose obese gene product, leptin, inhibits bovine ovarian thecal cell steroidogenesis Biology of Reproduction 58:207212 Thornton JE, Cheung CC, Clifton DK, Steiner RA (1997) Regulation of hypothalamic proopiomelanocrotin mRNA by leptin in ob/ob mice Endocrinology 138 50635065 Tomimatsu T, Yamaguci M, Murukami T et al. (1997) Increase of mouse leptin production by adipose tissue after midpregnancy: gestational prole of serum leptin concentration Biochemical and Biophysical Research Communications 240 213215

Foster DL, Bucholtz DC and Herbosa CG (1995) Metabolic signals and the timing of puberty in sheep. In The Neurobiology of Puberty pp 243257 Eds TM Plant and PA Lee. Journal of Endocrinology, Bristol Goldstone AP, Mercer JG, Gunn I et al. (1997) Leptin interacts with glucagon-like peptide-1 neurons to reduce food intake and body weight in rodents FEBS Letters 415 134138 Gonzalez MI, Baker BI and Wilson CA (1997) Stimulatory effect of melaninconcentrating hormone on luteinising hormone release Neuroendocrinology 66 254262 Gruaz NM, Lalaoui M, Pierroz DD, Englaro P, Sizonenko PC, Blum WF and Aubert ML (1998) Chronic administration of leptin into the lateral ventricle induces sexual maturation in severely food-restricted female rats Journal of Neuroendocrinology 10 627633 Hakansson M-L, Brown H, Ghilardi N, Skoda RC and Meister B (1998) Leptin receptor immunoreactivity in chemically dened target neurons of the hypothalamus Journal of Neuroscience 18 559572 Halaas JL, Gajiwala KS, Maffei M, Cohen SL, Chait BT, Rabinowitz D, Lallone RL, Burley SK and Friedman JM (1995) Weight-reducing effects of the plasma protein encoded by the obese gene Science 269 543546 Hardie L, Trayhurn P, Abramovich D and Fowler P (1997) Circulating leptin in women: a longitudinal study in the menstrual cycle and during pregnancy Clinical Endocrinology 47 101106 Harris RBS, Zhou J, Redmann SM, Smagin GN, Smith SR, Rodgers E and Zachwieja JJ (1998) A leptin doseresponse study in obese (ob/ob) and lean (=/?) mice Endocrinology 139 819 Henry B, Goding J, Alexander W, Tilbrook A, Canny B, Dunshea F and Clarke IJ (1998) Central administration of leptin to ovariectomised ewes inhibits food intake without affecting the secretion of hormones from the pituitary gland: evidence for a dissociation of effects on appetite and neuroendocrine function Endocrinology Hoggard N, Mercer JG, Rayner DV, Moar K Trayhurn P and Williams LM (1997) Localization of leptin receptor mRNA splice variants in murine peripheral tissues by RTPCR and in situ hybridisation Biochemical and Biophysical Research Communications 232 383387 Kalra SP (1993) Mandatory neuropeptide-steroid signalling for the preovulatory luteinizing hormone-releasing hormone discharge Endocrine Reviews 14 507538 Karlsson C, Lindell K, Svensson E, Bergh C, Lind P, Billig H, Carlsson LMS and Carlsson B (1997). Expression of functional leptin receptors in the human ovary Journal of Clinical Endocrinology and Metabolism 82 41444148 Kawai M, Yamaguchi M, Murukami T, Shima K, Murata Y and Kishi K (1997) The placenta is not the main source of leptin production in pregnant rat: gestational prole of leptin in plasma and adipose tissues Biochemical and Biophysical Research Communications 240 798802 Lee G, Proence R, Montez JM, Carroll KM, Darvishzadeh JG, Lee JI and Friedman JM (1996) Abnormal splicing of the leptin receptor in diabetic mice Nature (London) 379 632635 Linzer DIH and Arey BJ (1995) Ovarian prolactin receptors and their placental ligands. In The Somatotrophic Axis and the Reproductive Process in Health and Disease pp 2839 Eds EY Adashi and MO Thorner. Serono Symposia USA. Springer Verlag, New York Loucks AB and Heath EM (1994) Dietary restriction reduces luteinising hormone (LH) pulse frequency during waking hours and increases LH pulse amplitude during sleep in young menstruating women Journal of Clinical Endocrinology and Metabolism 78 910915 McGregor GP, Desaga JF, Ehlenz K, Heese F, Hegele A, Lammer C, Peiser C and Lang RE (1996) Radioimmunological measurement of leptin in plasma of obese and diabetic human subjects Endocrinology 137 15011504 Malik KF and Young S, III (1996) Localisation of binding sites in the central nervous system for leptin (OB protein) in normal, obese (ob/ob) and diabetic (db/db) C57BL/6J mice Endocrinology 137 14971500 Martin GB, Walkden-Brown SW, Boukhil R, Tjondronegoro S, Miller DW, Fisher JS, Hotzel M-J, Restall BJ and Adams NR (1994) Non-photoperiodic inputs into seasonal breeding in male ruminants. In Perspectives in Comparative Endocrinology pp 574585. Eds KG Davey, RE Peter and SS Tobe. National Research Council of Canada, Ottawa *Masuzaki H, Ogawa Y, Sagawa N et al. (1997) Nonadipose tissue production of leptin: leptin as a novel placenta-derived hormone in humans Nature Medicine 3 10291033

Leptin and reproduction

Van der Walt LA, Wilmsen EN and Jenkins T (1978) Unusual sex hormone patterns among desert-dwelling hunter-gatherers Journal of Clinical Endocrinology and Metabolism 46 658663 Weigle DS, Duell PB, Connor WE, Steiner RA, Soules MR and Kuijper JL (1997) Effect of fasting, re-feeding and dietary fat restriction on plasma leptin levels Journal of Clinical Endocrinology and Metabolism 82 561565 Yen SSC Lifestyle and body composition effects on the ovary. In Endocrinology and Metabolic Clinics of North America Eds HG Burger and RI McLachlan. WB Saunders, Philadelphia (in press)

55

Yu WH, Kimura M, Walczewska A, Karanth S and McCann SM (1997) Role of leptin in hypothalamicpituitary function Proceedings of the National Academy of Science USA 94 10231028 Zachow RJ and Magofn DA (1997) Direct intraovarian effects of leptin: impairment of the synergistic action of insulin-like growth factor I on follicle-stimulating hormone-dependent estradiol-17 production by rat ovarian granulosa cells Endocrinology 138 847850 Zamorano PL, Mahesh VB, De Sevilla LM, Chorich LP, Bhat GK and Brann DW (1997) Expression and localization of the leptin receptor in endocrine and neuroendocrine tissues of the rat Neuroendocrinology 65 223228