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J. J. JACOBS
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R. D. PEPPLER
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Department of Anatomy, Louisiana State University Medical Center, New Orleans, Louisiana 70112, U.S.A.

(Received 12 April 1977)

It is known that a functional interaction exists between the adrenal and ovarian systems, albeit complex and not well understood. Removal of progesterone derived from the adrenal gland disrupts the synchronous discharge of luteinizing hormone (LH) and consequently the events associated with reproduction (Mann, Korowitz & Barraclough, 1975). Adrenalectomy of female rats also delays the onset of puberty, disrupts the normal oestrous cycle and decreases the number of ova shed and the size of the litter (Ramaley, 1974). Previously we reported that unilaterally ovariectomized rats which had been bilaterally adrenalectomized for 30 days did not, after one oestrous cycle, show compensatory ovulation or ovarian hypertrophy; rats adrenalectomized and ovariectomized for 30 days did, however, display both phenomena (Peppler & Jacobs, 1976). While continuing these studies on adrenal gland–ovary interrelationships, we collected data which

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H. MORISHITA
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F. NAFTOLIN
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R. B. TODD
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R. WILEN
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I. J. DAVIES
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K. J. RYAN
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Department of Obstetrics and Gynecology, and Laboratory of Human Reproduction and Reproductive Biology, Harvard Medical School, Boston, Massachusetts 02115, U.S.A.

(Received 19 May 1975)

Testosterone and its ring A reduced metabolite, dihydrotestosterone (DHT), lower gonadotrophin concentrations in weaned rats (Swerdloff, Walsh & Odell, 1972; Naftolin & Feder, 1973). While neonatally administered testosterone causes anovulatory sterility in adult female rats, similar treatment with DHT has no apparent effect upon central neuroendocrine programming (Brown-Grant, Munck, Naftolin & Sherwood, 1971; Whalen & Luttge, 1971), which raises the question of whether DHT can suppress gonadotrophins in the neonatal rat.

In the first experiment, 5-day-old Sprague–Dawley-derived rats (Charles River Farms) were injected s.c. with 100 μg DHT propionate (DHTP), 100 μg testosterone propionate (TP, Eli Lilly & Co.) or the sesame oil diluent, and allowed to develop without further treatment. The injection volume was 25 μl. All animals were weaned on day 20. Daily vaginal

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M. J. BAUM
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P. J. M. SCHRETLEN
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Department of Endocrinology, Growth and Reproduction, Erasmus University, Faculty of Medicine, Rotterdam, The Netherlands

(Received 1 March 1978)

In several avian species the ability of exogenous oestrogen to induce nest-building and in some instances courtship behaviour after ovariectomy is significantly enhanced by exposing the birds to long environmental photoperiods (Steel & Hinde, 1972; Liley, 1976). The same lighting stimulus will also induce ovarian activity in intact birds and a related phenomenon has also been reported in the sheep (Raeside & McDonald, 1959; Fletcher & Lindsay, 1971). In the studies on sheep, it was reported that ovariectomized ewes primed with progesterone displayed sexual behaviour in response to treatment with oestradiol benzoate more readily during the normal breeding season, when days were short, than at other times of the year. This communication reports a failure to obtain comparable results in another seasonally breeding mammal, the ferret (Mustela furo).

Female ferrets were purchased

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J Naufahu Faculty of Science and Technology, University of Westminster, London, UK

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F Alzaid Faculty of Science and Technology, University of Westminster, London, UK

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M Fiuza Brito Faculty of Science and Technology, University of Westminster, London, UK

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B Doslikova Faculty of Science and Technology, University of Westminster, London, UK

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T Valencia Faculty of Science and Technology, University of Westminster, London, UK

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A Cunliffe Faculty of Science and Technology, University of Westminster, London, UK

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J F Murray Faculty of Science and Technology, University of Westminster, London, UK

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-sectional study Fasting venous blood samples were collected from 135 females and 96 males. Weight to the nearest 0.1 kg, height to the nearest 0.1 cm, waist and hip circumference were measured. Total fat and lean body mass were measured by air displacement

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DP de Kleijn
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KP Janssen
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SL Waddy
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R Hegeman
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WY Lai
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GJ Martens
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F Van Herp
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Crustacean reproduction is regulated by a complex chain of hormonal interactions in which the crustacean hyperglycaemic hormones A and B (CHH-A and CHH-B) and the gonad-inhibiting hormone (GIH) play a primary role. These neurohormones are produced in the same neuroendocrine cells of the X-organ sinus gland complex, situated in the eyestalks of the American lobster, Homarus americanus. In order to obtain more information on the synthesis, storage, release and function of these three neuropeptides during the reproductive cycle, we studied the levels of their mRNAs in the X-organ, their peptide storage in the sinus gland and their concentration in the haemolymph at different stages of the female reproductive cycle. A high CHH-A mRNA level was found only in the previtellogenic stage, while elevated mRNA levels were determined for CHH-B in the mature as well as the previtellogenic stage. High CHH storage levels in the sinus gland were found during previtellogenesis. The total amount of CHH (CHH-A plus -B) in the haemolymph was significantly higher during maturation. A low level of GIH mRNA in the X-organ and a low amount of the GIH I isoform in the sinus gland were found only in the immature stage. In contrast, GIH haemolymph levels were high during the immature and previtellogenic stages. We conclude that CHH-A and -B are involved in triggering the onset of vitellogenesis and that CHH-B in particular is responsible for stimulating oocyte maturation before spawning, while GIH prevents the start of vitellogenesis in the ovary. Moreover, our results show that the balance between the haemolymph levels of the CHHs and GIH may tune the synchronization of reproduction and molting during the biannual reproductive cycle of the American lobster.

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D Blache
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RL Tellam
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LM Chagas
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MA Blackberry
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PE Vercoe
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GB Martin
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In mature male sheep, the level of nutrition acutely influences the secretion of reproductive hormones. The mechanism involved is not fully understood but findings in humans and laboratory rodents would suggest a major role for leptin that is secreted from adipose tissue and then travels via the circulation to the central nervous system. Before we can begin to test this hypothesis, we need to be able to measure leptin concentrations in blood plasma and cerebrospinal fluid. We have therefore developed a radioimmunoassay using antibodies raised against biologically active recombinant bovine-ovine leptin. Using this assay, we found that plasma concentrations of leptin were highly correlated to back-fat thickness and to the ratio of back-fat thickness to liveweight, in female and castrated male sheep. Plasma concentrations of leptin were higher in female sheep than in castrated or intact male sheep. Serial samples (every 5 min) suggested that the secretion of leptin in male sheep is episodic but it does not appear to show clear pulsatility, increases post-prandially, or a diurnal rhythm. Leptin concentrations in both plasma and cerebrospinal fluid increased within 5 days in male sheep fed a diet with a high content of energy and protein that also stimulates the secretion of LH pulses. These data suggest that in sheep, as in other species, leptin production is correlated with the mass of adipose tissue and that the hormone passes from the circulation to the cerebrospinal fluid and then to hypothalamic sites. There, it may affect appetite and perhaps GnRH secretion. The role of leptin in the link between nutrition and reproduction needs further investigation.

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KL Britt
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JK Findlay
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Estrogens are synonymous with fertility and infertility in mammals. Our knowledge of the biological actions of estrogens, however, is incomplete. Three recent developments have thrown new light on the actions of estrogens in mammalian reproduction that will lead to a greater understanding of their functions. They are (a) the identification of a second estrogen receptor, called ERbeta, (b) the identification of ligand-specific ER coactivators and (c) mouse models with targeted disruption of the genes encoding both ER and the aromatase enzyme. These models provide for the first time animals which are either unable to respond to endogenous or exogenous estrogens (ER 'knockouts'), or can respond to exogenous estrogen but do not make endogenous estrogen (aromatase 'knockout' or ArKO). Furthermore, the ArKO mouse has provided a model to study the effects on the ovary of exogenous estrogens of plant and synthetic origin that are of clinical relevance. The data show that estrogens are essential for fertility but not for survival after birth or for the formation of the reproductive tract. This commentary focuses on the roles of estrogen in folliculogenesis and in the maintenance of the ovarian somatic cell phenotype in the mouse. We also hypothesize that the ERalpha and ERbeta may subserve the proliferative and differentiative actions of estrogen, respectively, within a follicle. In summary, estrogen is obligatory for normal folliculogenesis beyond the antral stage and for the maintenance of the female phenotype of the somatic cells within the ovaries. This clearly demonstrates a major role for sex steroids in somatic cell differentiation in the gonads of eutherian mammals and challenges the central paradigm that the ovary is the default gonad, arising due to the absence of testicular defining signals. Evidence is also provided for the plasticity of the adult female gonad. Understanding the mechanisms of estrogen actions will provide an insight into the regulation of reproductive disorders afflicting women today, notably ovarian dysfunction and the menopause.

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M. Rand-Weaver
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P. Swanson
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H. Kawauchi
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W. W. Dickhoff
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ABSTRACT

Somatolactin (SL), a newly discovered fish pituitary protein belonging to the GH/prolactin family, was isolated from coho salmon (Oncorhynchus kisutch). Antibodies were raised to purified coho SL, and a homologous radioimmunoassay was developed and validated. The assay was specific for SL as indicated by the absence of cross-reactivity with coho salmon GH, gonadotrophins I and II and less than 0·2% cross-reaction to prolactin. Serial dilutions of plasma and pituitary extracts from Oncorhynchus species including coho salmon, chinook salmon and rainbow trout were parallel to the coho salmon SL standard curve. Displacement curves for dilutions of Atlantic salmon (Salmo salar) plasma, but not pituitary extract were parallel to the standards. Plasma levels of SL were measured in coho salmon throughout the final year of reproductive maturation. During the period of gonadal growth, plasma SL levels increased and were highly correlated to oestradiol levels in females and 11-ketotestosterone levels in males. Peak levels of SL were observed at the time of final maturation and spawning in both sexes. It is hypothesized that SL may regulate some physiological aspect of reproduction.

Journal of Endocrinology (1992) 133, 393–403

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K Toda
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T Okada
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K Takeda
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S Akira
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T Saibara
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M Shiraishi
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S Onishi
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Y Shizuta
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Aromatase P450 (CYP19) is an enzyme responsible for the conversion of androgens to oestrogens. We generated CYP19 knockout (ArKO) mice by targeted disruption of Cyp19 and studied the role of oestrogens in male reproductive ability. Approximately 85% of ArKO males were unable to sire offspring. However, no obvious difference was found in testicular and epididymal weights, numbers of sperm in the epididymis or the ability of sperm to fertilize eggs in vitro between wild-type and ArKO males. An examination of mating behaviour demonstrated that ArKO males showed an impairment in mounting behaviour against sexually mature females. The inability of more than 90% of ArKO males to sire offspring was reversed by repeated subcutaneous injections of 17beta-oestradiol when initiated on the day of birth. The effects of 17beta-oestradiol on reproduction were concentration dependent and evident when supplementation was initiated on day 7, but not on day 15 after birth. These findings suggest that oestrogens acting during neonatal life are required for normal mating behaviour in adulthood.

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S Kajimura
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N Kawaguchi
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T Kaneko
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I Kawazoe
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T Hirano
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N Visitacion
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EG Grau
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K Aida
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There is considerable evidence that the GH/IGF-I axis plays an important role in female reproduction. We report the isolation and characterization of the GH receptor (GH-R) and its gene expression profile during oogenesis in the tilapia, Oreochromis mossambicus. cDNA encoding GH-R was cloned and sequenced from the tilapia liver. The predicted GH-R preprotein consisted of 635 amino acids and contained a putative signal peptide, an extracellular region with a characteristic motif, a single transmembrane region, and a cytoplasmic region with conserved box 1 and 2 domains. The tilapia GH-R shared 34-74% identities with known GH-Rs in vertebrates. A binding assay using COS-7 cells showed that the cloned GH-R bound specifically to tilapia GH. Northern blot analysis showed a single mRNA transcript in the liver and ovary. In situ hybridization revealed intense signals of GH-R in the cytoplasm and nucleus of immature oocytes. The granulosa and theca cells surrounding vitellogenic oocytes also contained the GH-R mRNA signals. About a tenfold greater level of GH-R mRNA was found in the immature oocytes versus the mature oocytes, along with high levels of IGF-I mRNA. There were no significant changes in mRNA levels of GH-R and IGF-I in the liver or in plasma IGF-I levels during oocyte development. No correlation was found between hepatic GH-R mRNA and ovarian GH-R mRNA. These results suggest that the GH/IGF-I axis in the ovary may be involved in the early phases of oogenesis, under a different regulatory mechanism of GH-R gene expression from that of the liver.

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