To clarify the participation of insulin-like growth factor type-II (IGF-II) in rat gonadal differentiation, immunohistochemical localization of IGF-II was chronologically studied in Sprague-Dawley rat gonads from gestational day (GD) 13 to postnatal day (PD) 21 by using avidin-biotin complex technique. In male gonads, most cells were negative to IGF-II immunostaining during the perinatal period. Only Leydig/interstitial cells expressed positive reactivity from GD 21 to PD 11: the intensity of staining and the number of positive cells were gradually increased until PD 11. In female gonads, almost all cells showed negative immunoreactivity. Mesonephric tubules in both sexes exhibited slight or moderate reactivity on GD 13. These results indicate that IGF-II is likely to participate in the regression of fetal-type Leydig cells and/or the proliferation of adult-type Leydig cells around birth, and in the development of mesonephric tubules in the initial stage of gonadal differentiation.
It is well established that enhancement of RNA polymerase activity is one of the earlier androgenic effects in the rat ventral prostate. Attention has been given to selective binding of androgen by prostatic chromatin where it may influence the synthesis of RNA (Steggles, Spelsberg, Glasser & O'Malley, 1971), and the relationship between androgen and the nuclear RNA synthesizing system in the prostate has been the subject of intensive research. In the present study we report the effects of androgens and the prostatic cytoplasm containing androgen receptor on RNA synthesis of isolated prostatic nuclei. Male adult rats were used in the following experiments 3 days after castration. Nuclei from rat ventral prostates were prepared as described by Liao, Leininger, Sagher & Barton (1965). Tissue was incubated in 0·35 ml medium containing 0·4 μmol each of GTP, CTP, and ATP, 0·8 μCi [3H]UTP (17 Ci/mmol), 1·2 μmol β-mercaptoethanol, 35 μmol KCl,
Previously we examined that inhibin-alpha subunit, transforming growth factor-beta 1 (TGF-beta 1) and epidermal growth factor (EGF) were expressed in sex-, cell- and stage-specific manners in perinatal rat gonads. To clarify effects of these growth factors on the rat gonadal differentiation and development, indifferent gonadal primordia with mesonephric tubules on gestational day 13 were cultured in vitro for 4 days in serum-free CMRL-1066 medium with inhibin, TGF-beta 1, EGF, anti-sera against these growth factors, testosterone or estradiol-17 beta, and then morphologically examined with reference to seminiferous tubule formation, germ cell division, Wolffian and Müllerian duct development. In male gonads, anti-inhibin-alpha serum suppressed the seminiferous tubule formation but inhibin, TGF-beta 1, EGF or steroid hormones did not affect on the tubule formation, germ cell proliferation nor gonoduct development. Seminiferous tubules in male gonads cultured in the medium containing anti-inhibin-alpha serum were incomplete and irregular in shape. On the other hand, in female gonads, inhibin suppressed the germ cell division and anti-inhibin-alpha serum led to the necrosis of germ cells, but other factors affected to neither sex cord formation nor germ cell division. Testosterone and estradiol-17 beta stimulated female Wolffian and Müllerian duct development, respectively. These results indicate that inhibin induces the seminiferous tubule formation and suppresses the female germ cell division in developing rat gonads in vitro.
The influence of castration on the incorporation of nucleotide precursors into RNA of isolated prostatic tissue has been investigated. Castration brought about an apparent increase of incorporation by increasing the specific activity of the uridine nucleotide pool and not by an enhancement in synthesis of rapidly labelled RNA. An actual decrease in this reaction was shown by dividing the radioactivity incorporated into RNA by the specific activity of the uridine nucleotide. Administration of testosterone brought about enhanced synthesis of RNA in the prostates of rats castrated for 3 days as early as 4 h after injection, reaching maximum effect at 8 h. No significant difference was found between the turnover rates of rapidly labelled RNA from castrated, androgen-treated castrated and intact rats. The increased prostatic permeability to nucleosides as an early action of androgens in this organ is discussed.
