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Differential localization of inhibin subunit proteins in the ovine testis during fetal gonadal development.

Inhibins and activins are dimeric proteins that are involved in cell proliferation, apoptosis, and differentiation in a number of systems and have previously been detected in fetal testes of many species. This study used immunohistochemistry to examine the localization of inhibin alpha-, betaA-, and betaB- subunits during ovine testicular development from days 40-135 of gestation. Localization of inhibin betaA- and betaB-subunit messenger RNAs was confirmed by in situ hybridization. The results showed that there was differential localization of inhibin alpha-, betaA-, and betaB-subunits to specific cells in the ovine fetal testis from 40 days of gestation. All three inhibin subunits were present in Sertoli cells throughout gestation, whereas the rete epithelium and gonocytes did not express inhibin alpha-subunit. These data suggest that the fetal Sertoli cells have the capacity to produce all forms of inhibins and activins, i.e. inhibin A and B, and activins A, AB, and B, whereas the rete testis epithelial cells can only synthesize activin A. In the interstitium, the fetal Leydig cells expressed all three inhibin subunits, but this was restricted to the period between 40 and 90 days of gestation. Thereafter, inhibin alpha-subunit immunoreactivity was not observed in fetal Leydig cells, which suggests that only activin ligands are produced by Leydig cells during late gestation. Collectively, the data demonstrate that fetal ovine testes have the potential to produce the full repertoire of inhibins and activins from very early in testicular differentiation. The distinct and restricted localization of the various subunits to specific cells suggests that specific dimeric proteins have particular roles in the development and function of the fetal testis.

Animals↗

Dax-1 (dosage-sensitive sex reversal-adrenal hypoplasia congenita critical region on the X chromosome, gene 1) gene transcription is regulated by wnt4 in the female developing gonad.

Dax-1 [dosage-sensitive sex reversal-adrenal hypoplasia congenita critical region on the X chromosome, gene 1 (NR0B1)] is an orphan nuclear receptor acting as a suppressor of Ad4 binding protein/steroidogenic factor 1 [Ad4BP/SF-1 (NR5A1)] and as an anti-Sry factor in the process of gonadal sex differentiation. The roles of these nuclear receptors in the differentiation of the gonads and the adrenal cortex have been established through studies of the mutant phenotype in both mice and humans. However, the mechanisms underlying transcriptional regulation of these genes remain largely unknown. Here, we examined the relationship between Dax-1 gene transcription and the Wnt4 pathway. Reporter gene analysis revealed that Dax-1 gene transcription was activated by beta-catenin, a key signal-transducing protein in the Wnt pathway, acting in synergy with Ad4BP/SF-1. Interaction between beta-catenin and Ad4BP/SF-1 was observed using yeast two-hybrid and in vitro pull-down assays. The region of Ad4BP/SF-1 essential for this interaction consists of an acidic amino acid cluster, which resides in the first helix of the ligand-binding domain. Mutation of the amino acid cluster impaired transcriptional activation of Dax-1 as well as interaction of Ad4BP/SF-1 with beta-catenin. These results were supported by in vivo observations using Wnt4 gene-disrupted mice, in which Dax-1 gene expression was decreased significantly in sexually differentiating female gonads. We thus conclude that Wnt4 signaling mediates the increased expression of Dax-1 as the ovary becomes sexually differentiated.

Amino Acids↗

Gonadal development and gonadotrophin secretion in the male vole (Microtus agrestis) after an abrupt change in photoperiod.

Male voles were reared from birth to age 28 days in 6L:18D. Pairs of animals showing similar sexual development were assigned at random to 16L:8D or 6L:18D. Treatments continued for a further 56 days. Increase in the activity of the hypothalamo-hypophysial system occurred within 4 days of exposure to 16L:8D, as shown by significant elevation of plasma LH and FSH. Pituitary LH did not increase until Day 7, and pituitary FSH did not increase until Day 21. After exposure to 16L:8D for 4 days, pituitary FSH was lower than in corresponding animals in 6L:18D. These discrepancies between pituitary and plasma values of gonadotrophins indicate that increase in hormone release occurs before synthesis is fully stimulated. Enhanced output of testicular hormones probably began between Day 7 and Day 14, as indicated by an increase in seminal vesicle weight, yet plasma and pituitary concentrations of LH and FSH remained elevated. This suggests that long photoperiods may cause direct stimulation of the hypothalamo-hypophysial system which increasing values of testicular hormones are initially unable to inhibit. The response of this system in voles to an abrupt change from a non-stimulating to a stimulating photoperiod has a time course resembling that for the Soay ram but appreciably slower than for the Japanese quail.

Animals↗

Effects of a gonadotrophin-releasing hormone antagonist on gonadotrophin secretion and gonadal development in neonatal pigs.

Male (N = 8) and female (N = 8) pigs were assigned to receive saline or a potent GnRH antagonist ([Ac-D2Nal1,D4-Cl-Phe2,D-Trp3,D-Arg6, D-Ala10]- GnRH*HOAc; 1 mg/kg body weight) at 14 days of age. The GnRH antagonist caused LH to decline (P less than 0.01) from 1.7 ng/ml at 0 h to less than 0.5 ng/ml during 4-32 h in males and females. Concentrations of FSH in gilts declined slowly from 75 +/- 8 to 56 +/- 5 ng/ml (P less than 0.05) at 32 h. In males FSH was low (5.7 +/- 0.5 ng/ml) at 0 h and did not change significantly. To observe the effect of long-term treatment with GnRH antagonist, 10 male and 10 female pigs, 3 days of age, were treated with saline or 1 mg GnRH antagonist per kg body weight every 36 h for 21 days. Concentrations of LH were reduced (P less than 0.01) to 0.2-0.4 ng/ml throughout the experimental period in male and female piglets treated with GnRH antagonist. Plasma FSH increased in control females, but remained suppressed (P less than 0.001) in females treated with GnRH antagonist. Treatment with the GnRH antagonist suppressed FSH levels in males on Days 8 and 16 (P less than 0.05), but not on Day 24. Treatment of females with the GnRH antagonist did not influence (P greater than 0.10) oestradiol-17 beta concentrations. Administration of GnRH antagonist to males suppressed testosterone and oestradiol-17 beta values (P less than 0.01) and reduced testicular weight (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Postnatal growth rate and gonadal development in circadian tau mutant hamsters reared in constant dim red light.

The role of the circadian clock in the reproductive development of Syrian hamsters (Mesocricetus auratus was examined in wild type and circadian tau mutant hamsters reared from birth to 26 weeks of age under constant dim red light. Testis diameter and body weights were determined at weekly intervals in male hamsters from 4 weeks of age. In both genotypes, testicular development, subsequent regression and recrudescence exhibited a similar time course. The age at which animals displayed reproductive photosensitivity, as exhibited by testicular regression, was unrelated to circadian genotype (mean +/- SEM: 54 +/- 3 days for wild type and 59 +/- 5 days for tau mutants). In contrast, our studies revealed a significant impact of the mutation on somatic growth, such that tau mutants weighed 18% less than wild types at the end of the experiment. Our study reveals that the juvenile onset of reproductive photoperiodism in Syrian hamsters is not timed by the circadian system.

Analysis of Variance↗

Gonad development and gametogenesis in Boophilus decoloratus (Koch, 1844) (Acarina Metastriata Ixodidae).

Phases in the growth of the reproductive organs and gametogenesis were followed in both males and females of Boophilus decoloratus (Koch, 1844). Special attention was given to the timing of meiosis, the production of mature spermiophores, mating and spermiophore relocation in the female. The main growth phase of the male testes takes place during the nymphal stage while that of the accessory gland complex takes place during the 4 days after moulting and before mating. The main growth phase of female ovaries and oviducts takes place during the post-mating period on the host and the previposition period. Meiosis in males takes place only after the attachment of the newly-moulted ticks; spematogenesis and spermiogenesis occur within a period of 4 days, after which pairing and spermatophore transfer take place. Meiosis was not observed in female ticks and it is concluded that this reduction division takes place only after spermiophore penetration of the oocytes and egg shell formation. Female ticks may be impregnated more than once, a maximum of 5 matings being deduced from counts of spermiophore capsules in the female seminal receptacle. Each spermatophore produced by male ticks yields either 1 but more commonly 2 spermiophore capsules. The reproduction of B. decoloratus is briefly discussed and compared with other Ixodoidea.

Animals↗