Transrectal ultrasonography of ovaries was performed each day in non-prolific Western white-faced (n = 12) and prolific Finn ewes (n = 7), during one oestrous cycle in the middle portion of the breeding season (October-December), to record the number and size of all follicles > or = 3 mm in diameter. Blood samples collected once a day were analysed by radioimmunoassay for concentrations of LH, FSH and oestradiol. A cycle-detection computer program was used to identify transient increases in concentrations of FSH and oestradiol in individual ewes. Follicular and hormonal data were then analysed for associations between different stages of the lifespan of the largest follicles of follicular waves, and detected fluctuations in serum concentrations of FSH and oestradiol. A follicular wave was defined as a follicle or a group of follicles that began to grow from 3 to > or = 5 mm in diameter within a 48 h period. An average of four follicular waves per ewe emerged during the interovulatory interval in both breeds of sheep studied. The last follicular wave of the oestrous cycle contained ovulatory follicles in all ewes, and the penultimate wave contained ovulatory follicles in 10% of white-faced ewes but in 57% of Finn ewes. Transient increases in serum concentrations of FSH were detected in all animals and concentrations reached peak values on days that approximated to follicle wave emergence. Follicular wave emergence was associated with the onset of transient increases in serum concentrations of oestradiol, and the end of the growth phase of the largest follicles (> or = 5 mm in diameter) was associated with peak serum concentrations of oestradiol. Serum FSH concentrations were higher in Finn than in Western white-faced ewes during the follicular phase of the cycle (P < 0.05). There were no significant differences in serum concentrations of LH between Western white-faced and Finn ewes (P > 0.05). Mean serum concentrations of oestradiol were higher in Finn compared with Western white-faced ewes (P < 0.01). It was concluded that follicular waves (follicles growing from 3 to > or = 5 mm in diameter) occurred in both prolific and non-prolific genotypes of ewes and were closely associated with increased secretion of FSH and oestradiol. The increased ovulation rate in prolific Finn ewes appeared to be due primarily to an extended period of ovulatory follicle recruitment.

In this review, we describe the process of sexual maturation in the bull calf. The testes of the bull grow relatively slowly until approximately 25 weeks of age and then a rapid phase of growth occurs until puberty, at 37-50 weeks of age. During the early postnatal phase of slower growth of the testis pre-spermatogonia and some spermatogonia are established, adult Leydig cells appear and undifferentiated Sertoli cells are produced. The rapid testicular growth, after 25 weeks of age, consists of marked increases in the diameter and length of the seminiferous tubules, dramatic proliferation and differentiation of germ cells, with mature spermatozoa occurring between 32 and 40 weeks of age. The adult Leydig cell population is largely in place by 30 weeks of age and that of Sertoli cells by 30-40 weeks of age. Serum concentrations of LH increase from 4 to 5 weeks of age, to an early postnatal peak at 12-16 weeks of age, followed by a decline to 25 weeks of age. Serum FSH concentrations are high postnatally, declining to approximately 25 weeks of age. Serum testosterone concentrations increase during the phase of rapid testicular growth. Hypothalamic opioidergic inhibition may abate transiently to allow the early postnatal increase in LH secretion, while testicular androgenic negative feedback probably contributes to the decline in gonadotropin secretion to 25 weeks of age. Several lines of study have led us to suggest that early postnatal gonadotropin secretion is pivotal in initiating the process of sexual maturation in the bull calf.

Data on pubertal maturation in male marmoset, a model for human reproduction, are scant and conflicting. We collected data on novel parameters to characterize puberty. Twenty-five marmoset monkeys were assigned to five age groups by weeks (wk): 21 (pre-pubertal), 43 (onset of puberty), 52 (fully pubertal), 70 (mature), and 116 (fully adult). Serum and intratesticular testosterone and pituitary bioactive chorionic gonadotropin (bioCG) were measured. Testicular development was assessed by ultrasonography, histology, and flow cytometry. Three consecutive blood samples revealed extreme fluctuations in testosterone concentrations, suggesting an erratic secretion. Age-related changes in serum testosterone and pituitary bioCG concentrations were observed. Intratesticular androgens (ITAs) showed high fluctuations within groups at all ages and were high in some animals by 21 wk. Unexpectedly, no correlation between pituitary bioCG and serum testosterone or ITAs was found, but these parameters significantly correlated with testicular weight and volume. These observations were consistent a dependence on the testis growth on bioCG. Unfortunately, the low serum levels of bioCG were not measurable in this study. At 43 wk, the animals reached puberty. At 52 wk of age, animals attained maximum body and epididymal weights and qualitatively normal spermatogenesis, but testes continued growing, reaching a maximum of all parameters at 70 wk of age, without further major changes at the age of 116 wk. It is concluded that (1) gonadal activation is evident at wk 21, (2) the male marmoset reaches the pubertal threshold around 43 wk of age, attains qualitative parameters at 52 wk, matures further to sexual maturity at 70 wk, and (3) serum testosterone and ITAs are highly variable without any identifiable correlation with pituitary bioCG.

The pituitary gonadotrophins LH and FSH are responsible for regulation of gametogenesis in the testis and ovary. Chorionic gonadotrophin (CG), a third closely related glycoprotein hormone derived by gene duplication of the LH gene and secreted by the placenta in primates, is essential for the rescue of the corpus luteum and maintenance of pregnancy. We have recently shown that marmoset (m) CG mRNA is highly expressed in the pituitary of the common marmoset (Callithrix jacchus) and that LH is less active than human CG in activating the human LH receptor lacking exon 10. To investigate further which gonadotrophin is the actual ligand of the LH receptor (LHR) of the marmoset monkey that naturally lacks exon 10, we identified and characterised the genomic organisation of the mLH gene and its expression. Intergenic PCR amplification of the region encompassing the mLH and the mCG genes revealed that, surprisingly, mCG is located 20 kbp upstream of the LH gene, whereas in other species the intergenic distance is approximately 2-3 kbp. Sequence analysis of the mLH coding region showed 70% identity to mCG and 90% identity to human LH at the amino acid level. Both gonadotrophin subunits are present at the genomic level, but RT-PCR of pituitary and placental total RNA using specific oligonucleotides for mCG and mLH showed high expression of mCG mRNA in both tissues, whereas LH was expressed neither in the pituitary nor in the placenta. Thus mLH mRNA is lacking in the marmoset pituitary. Immunohistochemistry of marmoset pituitaries showed that mCG was confined to the gonadotrophes, and partly co-localised in cells stained positively for FSH. Western blot analysis confirmed the presence of mCG in the pituitary. Northern blot analysis using mCG as a probe displayed one transcript of 0·7 kb in the pituitary and detected two transcripts of 1·1 kb and 2 kb in the marmoset placenta. Our results suggest that, in the common marmoset, CG is the only gonadotrophin with luteinising function that is present in the pituitary. We postulate that, owing to an unknown mutational event in evolution, expression of mLH was completely abolished, and CG -which, unlike LH, acts normally even when exon 10 is missing from the LHR -took over its function.

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Abstract. The effects of chronic blockade of androgen action by the antiandrogens flutamide and Casodex on serum and pituitary concentrations of LH and FSH, serum and testicular androgen levels, reproductive organ weights, and on spermatogenesis were compared in the adult rat. Animals were treated for 3 and 8 weeks with vehicle, Casodex (20 mg · kg−1 · (day)−1, flutamide (20 mg · kg−1 · (day)−1) and GnRH antagonist (150 μg/day, Detirelix). Treatment with GnRH antagonist suppressed gonadotropin and testosterone production, reduced the weights of testes, epididymides and seminal vesicles, and inhibited germ cell development. Flutamide administration markedly elevated serum and pituitary levels of gonadotropins as well as serum and testicular androgen concentrations. Casodex-induced elevation of gonadotropin concentrations was less pronounced and serum and testicular levels of androgens did not change significantly. The reduction of seminal vesicle weights was similar after Casodex and GnRH antagonist treatment, whereas flutamide was less effective. Testicular weight and spermatogenesis (assessed by light microscopical and flow-cytometric analysis) remained unaffected by Casodex and flutamide. It is concluded, that 1. Casodex, in contrast to flutamide, is a peripherally selective antiandrogen, and 2. Casodex influences release of gonadotropins into circulation less than flutamide. Therefore this antiandrogen might be useful clinically for selectively blocking androgen actions in the accessory sex glands.

Experiments were performed to evaluate the role of transcription in early development of bovine embryos. Two transcription inhibitors-5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) and actinomycin D-were used to test whether 1) the inhibitors alter the rate of early embryonic development and protein synthesis, 2) heat shock increases the steady-state amounts of mRNA for the inducible form of heat shock protein 70 (HSP70) in embryos, and 3) this latter effect is blocked by transcription inhibitors. Addition of either DRB or actinomycin D to culture medium beginning 8 h postinsemination (hpi) reduced the proportion of oocytes that had undergone cleavage by 32-34 hpi. Both transcription inhibitors also reduced the proportion of cleaved embryos that reached the 4-cell stage by 32-34 hpi. Incorporation of (35)S-labeled amino acids into de novo synthesized protein by bovine 2-cell embryos was lower for embryos cultured with DRB. Using reverse transcription-polymerase chain reaction, HSP70 mRNA in 2- and 4-cell embryos was increased by exposure to 42 degrees C. Both inhibitors reduced amounts of HSP70 mRNA at 42 degrees C. Results indicate that bovine embryos can undergo transcription in response to heat shock as early as the 2-cell stage. Moreover, the observations that transcription inhibitors reduce rates of cleavage and early development point out the importance of transcription for development from the earliest period of embryonic life.

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