Differential response patterns of kisspeptin and RFamide-related peptide to photoperiod and sex steroid feedback in the Djungarian hamster (Phodopus sungorus).

Many animals synchronise their reproductive activity with the seasons to optimise the survival of their offspring. This synchronisation involves switching on and off their gonadotrophic axis. Ever since their discovery as key regulators of gonadotrophin-releasing hormone (GnRH) neurones, the hypothalamic RF-amide peptides kisspeptin and RFamide-related peptide (RFRP) have been a major focus of research on the seasonal regulation of the gonadotrophic axis. In the present study, we investigated the regulation of both neuropeptides in the Djungarian hamster, a major animal model for the study of seasonal reproduction. During the long-day breeding period, kisspeptin neurones in the anteroventral periventricular area are solely controlled by a positive sex steroid feedback and, in the arcuate nucleus, they are subject to a very strong negative sex steroid feedback associated with a minor photoperiodic effect. During short-day sexual quiescence, the disappearance of this hormonal feedback leads to high levels of kisspeptin in arcuate neurones. Notably, chronic central administration of kisspeptin is able to over-ride the photoperiodic inhibition of the gonadotrophic axis and reactivate the reproductive function. Therefore, our data suggest that kisspeptin secretion by arcuate neurones during sexual quiescence is inhibited by mechanisms upstream of kisspeptin neurones. RFRP expression is solely controlled by photoperiod, being strongly reduced in short days independently of the sex steroid feedback. Thus, kisspeptin and RFRP display contrasting patterns of expression and regulation. Upstream mechanisms controlling these neurones should be the focus of further studies on the roles of these RFamide neuropeptides in the seasonal control of reproduction.

Photoperiod history-dependent responses to intermediate day lengths engage hypothalamic iodothyronine deiodinase type III mRNA expression.

Perihypothalamic thyroid hormone signaling features prominently in the seasonal control of reproductive physiology. Triiodothyronine (T(3)) signaling stimulates gonadal development, and decrements in T(3) signaling are associated with gonadal regression. Type 3 iodothyronine deiodinase (DIO3) converts the prohormone thyroxine (T(4)) into biologically inactive 3,3',5'-triiodothyronine, and in long-day breeding Siberian hamsters exposure to long (LD) and short (SD) photoperiods, respectively, inhibit and stimulate hypothalamic dio3 mRNA expression. Reproductive responses to intermediate-duration photoperiods (IntD) occur in a history-dependent manner; IntDs are interpreted as inhibitory only when preceded by longer photoperiods. Because dio3 expression has only been evaluated under LD or SD photoperiods, it is not known whether hypothalamic dio3 encodes absolute photoperiod duration or the reproductive interpretation of photoperiod. Male Siberian hamsters with and without a prior history of LD were exposed to IntD photoperiods, and hypothalamic dio3 mRNA expression was measured 6 wk later. Hamsters with a LD photoperiod history exhibited gonadal regression in IntD and a marked upregulation of hypothalamic dio3 expression, whereas in hamsters without prior exposure to LD, gonadal responses to IntD were absent, and dio3 expression remained low. Patterns of deiodinase expression in hamsters maintained in chronic IntD photoperiods did not appear to reflect feedback effects of gonadal status. Hypothalamic expression of dio3 does not exclusively reflect ambient photoperiod, but rather the context-dependent reproductive interpretation of photoperiod. Neuroendocrine mechanisms that compare current and prior photoperiods, which permit detection of directional changes in day length, occur either upstream, or at the level, of hypothalamic dio3 expression.

Assessment of the contraceptive efficacy of the aqueous extract of Aegle marmelos Corr. leaves in male albino rats.

The present study was conducted to evaluate the contraceptive effect of an aqueous extract from the leaves of Aegle marmelos (AMLAq) on the reproductive organs of male rats with an emphasis on reversibility. Adult male rats were treated daily with different doses of AMLAq, i.e., 150, 300 and 600 mg/kg bw/day for 60 days. The data presented in this study demonstrate that the weight of the reproductive organs was reduced significantly in all the treatment groups. AMLAq induced a significant decrease in the sperm motility and sperm density of the Cauda epididymis and testes. The reduction in fertility was 50%, 85% and 100%, respectively, in the treatment groups. The testosterone level also significantly declined. Biochemical analysis of the reproductive tissues for sialic acid, protein, glycogen, fructose, ascorbic acid, acid and alkaline phosphatase indicated a significant decrease whereas testicular cholesterol level significantly increased indicating alterations in the biochemical milieu of the genital organs. Fertility and other effects gradually returned to control levels 120 days after cessation of treatment. No clinical signs of side effects on general metabolism were detected throughout the treatment, and after withdrawal, body weight gain was similar in all groups together with no alterations in the weight of vital organs', hematological and serological parameters.

The inhibitory effects on adult male reproductive functions of crude garlic (Allium sativum) feeding.

AIM: to investigate the effects of crude garlic on adult male rat reproductive functions. METHODS: Thirty male rats were divided into five groups: group 1 (untreated) and groups 2, 3, 4 and 5 were fed for 30 days with 5%, 10%, 15% and 30% crude garlic, respectively. Testes and accessory organs were weighed and some markers were assessed. Light and electron microscopy observations were also performed. RESULTS: A significant decrease was observed in the body weight of groups 4 (14%; P < 0.01) and 5 (20%; P < 0.01); of the prostate weight in group 5 (29.1%; P < 0.05) and of seminal vesicle weight in groups 3 (14.4%; P < 0.01), 4 (18.3%; P < 0.01) and 5 (27.3%; P < 0.01). In contrast, testis and epididymis weights were unchanged. In epididymis tissue, the alpha glucosidase activity and the spermatozoa density were unchanged. The treatment resulted in a significant decrease in testosterone serum levels in groups 3 (77.3%; P < 0.01), 4 (77.3%; P < 0.01) and 5 (90.9%; P < 0.01), associated with a significant increase in LH serum levels (P < 0.01). Testicular histology showed a dose-dependent increase in the percentage of empty seminiferous tubules. Moreover, testicular function was affected; a significant decrease in phosphatase acid activity (P < 0.01) and testosterone (P < 0.05) contents were observed. CONCLUSION: Crude garlic consumption during 1 month reduced testosterone secretion and altered spermatogenesis at 10%, 15% and 30% doses.

Pueraria mirifica, a phytoestrogen-rich herb, prevents bone loss in orchidectomized rats.

OBJECTIVE: Estrogens and estrogen-like substances have been reported to play an important role in male bone homeostasis and to prevent bone loss. Pueraria mirifica (Leguminosae), a Thai herbal plant, containing a high amount of phytoestrogens was a choice of interest for this study. We examined the effects of crude P. mirifica on bone loss and influences on reproductive organs in male rats. METHODS: Using fully mature and orchidectomized (ORX) rats, the effects of 0, 10, 100 and 1000 mg/kgB.W./day of P. mirifica and 0.1mg/kg B.W./day of 17 alpha-ethinylestradiol (a positive control) were evaluated on bone mineral density (BMD) and bone mineral content (BMC) measured with a peripheral Quantitative Computerized Tomography (pQCT) densitometry. RESULTS: Bone loss in trabecular and cortical bones of the various sites of axial bone (fourth lumbar vertebral body) and long bones (tibia and femur) after ORX was dose-dependently prevented by P. mirifica. The effects were specific on bone types and sites. The weights of the accessory sex organs, seminal vesicle and ventral prostrate gland, which significantly decreased after 3-month of ORX, were not altered by P. mirifica. CONCLUSION: The results suggest that P. mirifica treatment may be useful to prevent an osteoporosis in elderly hypogonadism subjects without influences on reproductive organs.

Dose-response effects of intermittent PTH on cancellous bone in hindlimb unloaded rats.

UNLABELLED: HLU suppressed bone formation and resulted in bone loss in the tibial metaphysis of 6-month-old male rats. A human therapeutic dose of intermittent PTH (1 microg/kg/day) prevented the skeletal changes associated with HLU. INTRODUCTION: Skeletal unloading of skeletally mature rats results in trabecular thinning in the proximal tibial metaphysis, which is in part caused by a decrease in bone formation. We examined the efficacy of PTH in preventing the detrimental skeletal effects that occur with hindlimb unloading (HLU). MATERIALS AND METHODS: Six-month-old male Fisher 344 rats were HLU and treated with vehicle or recombinant human PTH(1-34) at 1, 5, 20, or 80 microg/kg/day for 2 weeks. The bone response was measured by microCT analysis of bone structure, histomorphometric analysis of static and dynamic bone parameters, and Northern blot analysis of mRNA levels for bone matrix proteins. The PTH-treated HLU animals were compared with vehicle-treated HLU and pair-fed normal weight-bearing controls. RESULTS: Unloading resulted in a decrease in cancellous bone volume that was caused in part by a dramatic 83% decrease in bone formation. All dose rates (1-80 microg/kg/day) of human PTH(1-34) significantly increased bone formation rates compared with vehicle-treated HLU controls. There was a dose response, and the highest dose rate of the hormone increased bone formation compared with normal weight-bearing rats by 708% (p < 0.0001). The increases in bone formation were accompanied by increases in mRNA levels for type 1 collagen, osteocalcin, and osteonectin. Also, treatment with PTH resulted in increases in mineral apposition rate and double-labeled perimeter, but the latter was disproportionally increased at high dose rates. A therapeutic dose of PTH (1 microg/kg/day) prevented disuse-induced trabecular thinning, whereas high-dose PTH (80 microg/kg/day) increased trabecular thickness compared with normal weight-bearing rats. CONCLUSIONS: These findings reveal that administration of a therapeutic dose of PTH to HLU rats prevents the decrease in bone formation and trabecular thinning, whereas high dose rates of the hormone increase bone formation and trabecular thickness to values that exceed normal values.

Inhibitory effects of retinoic acids on androgen-dependent development of neonatal mouse seminal vesicles in vitro.

The effects of retinoic acids (RAs) on development of seminal vesicles (SVs) of neonatal mice were investigated in vitro. SVs from 0-day-old male mice were cultured for 2-6 days in serum-free, chemically defined medium containing transferrin and BSA supplemented with 5alpha-dihydrotestosterone (DHT; 10(-8) M) and insulin (10 microg/ml), alone and in combination. Before culture, SVs from 0-day-old mice consisted of an unbranched epithelium surrounded by mesenchyme. SVs cultured in medium with DHT plus insulin or DHT alone formed numerous epithelial branches after day 2 of culture, whereas epithelial branching did not occur in SVs cultured with insulin alone. All-trans-RA or 13-cis-RA (10(-9)-10(-6) M) added to medium containing DHT plus insulin or DHT alone inhibited epithelial branching in a dose-dependent manner. This inhibitory effect was reversible after removal of the retinoids from the medium on day 4 of culture. These RAs also decreased [3H]thymidine labeling indexes of both epithelium and mesenchyme of SVs cultured in medium with DHT plus insulin or DHT alone and inhibited the increase in their protein contents. 9-Cis-RA was less inhibitory than all-trans-RA or 13-cis-RA on epithelial branching, [3H]thymidine labeling indexes of epithelium and mesenchyme, and protein content of SVs cultured in medium with DHT and insulin. In the absence of DHT (insulin alone), all-trans-RA did not affect either the [3H]thymidine labeling indexes of epithelium and mesenchyme or the protein content of cultured SVs. Reverse transcriptase-PCR demonstrated strong expression of transcripts for mouse RA receptors (RARalpha, RARgamma, and RXRalpha), with lower levels of expression of RARbeta, RXRbeta, and RXRgamma in neonatal SVs. The present results indicate that RAs reversibly inhibit androgen-dependent development of neonatal mouse SVs, most likely through RARs.

Characterization of the interaction of the cervane alkaloid, imperialine, at muscarinic receptors in vitro.

The action of the cervane alkaloid, imperialine, has been assessed at M1, M2 and M3 receptors in functional assays and at M1, M2, M3 and putative M4 sites in binding studies. In functional studies, imperialine acted as a selective surmountable antagonist at M2 receptors in guinea-pig isolated atria and uterus (-log KB = 7.7 and 7.4, respectively), in comparison to M1 receptors in canine isolated saphenous vein (-log KB = 6.9) or M3 receptors in a range of guinea-pig isolated smooth muscles including ileum, trachea, fundus, seminal vesicle or oesophagus (-log KB = 6.6-6.8). In rat aorta, the -log KB value at the M3 receptor (5.9) was slightly, but significantly, lower. In competition radioligand binding studies, imperialine was also selective toward to M2 sites in rat myocardium (-log Ki = 7.2) with respect to M1 and M3 sites (rat cerebral cortex, rat submaxillary gland; -log Ki = 6.1 and 5.7, respectively). However, it did not significantly discriminate between rat cardiac M2 sites and putative M4 sites in rabbit lung (-log Ki = 6.9). Imperialine resembles the alkaloid himbacine in terms of its pharmacological profile at muscarinic receptor subtypes in that it acts as an M2 selective antagonist with respect to M1 or M3 sites. It may also provide a second, commercially available, antagonist with which to discriminate between M1 and M4 receptors.

Interactions of testosterone and short-photoperiod exposure on the neuroendocrine axis of the male Syrian hamster.

Exposure of adult male golden hamsters to short days (less than 12.5 h light/day) leads to suppression of gonadal function which is secondary to reductions in gonadotropin and prolactin (PRL) secretion. Short-day (SD) exposure also leads to a reduction in hypothalamic norepinephrine (NE) and dopamine (DA) metabolism and an increase in hypothalamic LHRH content which appears to be related to a decrease in LHRH release. To determine whether SD-induced changes in NE and DA metabolism are dependent or independent of changes in circulating testosterone (T) levels and thus possible mediators of photoperiod effects on gonadotropin secretion, the effects of castration and steroid replacement on hypothalamic amine metabolism were studied in male hamsters maintained under long or short photoperiod conditions. The presence of Silastic T-implants resulted in a greater suppression of LH and FSH in SD than in long-day (LD) hamster, but increased median eminence (ME) LHRH content in both groups. Exposure of castrate hamsters to short days led to a reduction of NE turnover in the ME and medial preoptic-suprachiasmatic area (MPOA) and a decrease in serum FSH levels. LH levels tended to be lower, but not significantly so. The decrease in ME NE turnover was potentiated by T replacement, but in the MPOA-SCN, T-implants reversed the effects of short days. NE turnover in the MBH was reduced by T in both LD and SD animals, but the effect was much greater in the SD animals. SD exposure also caused a decrease in ME DA metabolism that was reversed by T replacement.(ABSTRACT TRUNCATED AT 250 WORDS)

Release of neurophysin I and oxytocin by stimulation of the genital organs in bulls.

Oxytocin and bovine neurophysin I (bNpI) were estimated by radioimmunoassay in jugular vein plasma which was collected continuously from 18 bulls. No release of peptides was observed during successive matings with a cow in oestrus or during successive mountings on a cow with ejaculations into an artificial vagina. Stimulation with an electro-ejaculator or, to a smaller extent, massage of the seminal vesicles and ampullae per rectum caused an increase of oxytocin accompanied by a release of bNpI. It is speculated that the release of these peptides is due to stimulation of afferent pelvic nerves in the rectal wall. Basal molar ratios of bNpI/oxytocin in the plasma were highly variable, often showing a large excess of either bNpI or oxytocin. After the onset of peptide release induced by stimulation, molar ratios approached 1:1. This might indicate that hormone release is by exocytosis. Basal bNpI does not provide a good reflection of the oxytocin level.