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C Shaha

Publications and source records attributed to C Shaha.

31 records · Page 2Linked to original sources

The ontogeny of immunoreactive beta-endorphin in fetal, neonatal, and pubertal testes from mouse and hamster.

UNLABELLED: Derivatives of proopiomelanocortin (POMC), physicochemically similar to beta-endorphin and desacetyl alpha MSH, have been identified in adult testes, where these peptides were localized to Leydig cells. In the present study, the presence of immunostainable derivatives of POMC was established in fetal, neonatal, and pubertal testes with the unlabeled antibody peroxidase-antiperoxidase method. Specificity of staining was established by absorption of primary antisera with excess antigen. In the mouse, immunoreactive beta-endorphin was detectable in a few primitive interstitial cells on day 14 of gestation, the day after testicular differentiation. Thereafter, the number of immunostainable cells progressively increased throughout fetal life, so that at birth, they comprised 55% of the total interstitial cells. After birth, the number of immunostaining cells declined, so that they were only 12% of interstitial cells by 5 days of age. After 10 days of age, the number of immunopositive cells progressively increased, and by 40 days, interstitial cells showed intense staining comparable to that in adult mice. At 10 days of age, when the number of immunostainable cells was low, hCG treatment increased both the number and staining intensity of beta-endorphin-positive cells to those seen in adult testes. Antibodies directed against gamma MSH, a peptide within the N-terminal segment of POMC, also produced specific staining of fetal and adult interstitial cells in the mouse. In the hamster, the pattern of staining with anti-beta-endorphin in fetal, neonatal, and pubertal interstitial cells was similar to that observed in mice; the number and staining intensity of immunostainable cells increased during fetal life, declined after birth, and rose again at puberty. IN CONCLUSION: 1) the number and staining intensity of immunostainable interstitial cells have two peaks in mouse and hamster, at birth and after puberty; 2) the number and staining intensity of mouse interstitial cells can be increased by hCG; and 3) the development of immunostainable beta-endorphin activity correlates with the previously reported spontaneous and hCG-induced maturation of morphology and enzyme activities of Leydig cells.

Animals

Demonstration of immunoreactive beta-endorphin- and gamma 3-melanocyte-stimulating hormone-related peptides in the ovaries of neonatal, cyclic, and pregnant mice.

Antisera against proopiomelanocortin (POMC)-derived peptides have previously been employed to demonstrate immunostainable materials in the male reproductive tract and in the corpus luteum of rat ovary. The present study was designed to determine how the distribution of such stainable materials varies in mouse ovary as a function of the reproductive status of the animal. Peptide-like activities were localized with the unlabeled antibody peroxidase-antiperoxidase (PAP) technique in ovaries removed from mice during fetal and neonatal development, during different stages of estrous cycle, and during pregnancy, with antisera against beta-endorphin, gamma 3MSH, and an extended N-terminal portion of POMC (16 K). beta-endorphin-like activity was also quantified in ovarian extracts by RIA. Immunostainable beta-endorphin, gamma 3MSH, and 16 K fragment-like activities were present in ovaries of pregnant and normally cycling (but not immature) mice. Intense staining was found predominantly in the corpora lutea. Less intense staining was observed in the interstitium and in the following parts of large follicles: parietal granulosa, corona radiata, and cumulus oophorus. When neonatal mice were injected with hCG, immunostainable beta-endorphin-like material in the ovarian interstitial area increased. Treatment with PMSG increased staining in both secondary follicles and the interstitium. Immunoassayable beta-endorphin-like activity was twice as high (per g wet wt) at pregnancy as during the cycle. We conclude that peptides similar or identical to POMC and/or its components are present in ovarian cells and that the concentration of such material appears to be regulated by gonadotropins.

Aging

Do testicular opiates regulate Leydig cell function?

beta-Endorphin is believed to be synthesized in testicular Leydig cells. To gain more information about the role of this and other endogenous opioid peptides in the testis, opiate antagonists (naloxone and nalmefene, 100 micrograms/testis) were administered intratesticularly to hemicastrated adult rats. Leydig cell function was evaluated by measurement of serum testosterone and testosterone production in vitro. Estimation of androgen binding protein (rABP) was used as an index of Sertoli cell function. Serum testosterone was reduced significantly by intratesticular administration of naloxone and nalmefene in treated animals. Systemic administration of these antagonists had no effect at the doses used. Testes from treated animals incubated in vitro with or without hCG produced significantly less testosterone than vehicle-treated control testes. Hemicastration reduced rABP synthesis and secretion; however, treatment with opiate antagonists did not alter the amount of this protein in the serum or epididymides of these rats. These observations suggest that endogenous testicular opiates modulate testosterone secretion by Leydig cells.

Androgen-Binding Protein

A species difference between hamster and rat in the effect of oestrogens on growth of large preantral follicles.

Intact or hypophysectomized 23-day-old hamsters and rats were injected s.c. with 2 mg diethylstilboestrol (DES) or 1 mg oestradiol cyclopentylpropionate (OECP) on Days 23-25 and killed on Day 26. Although serum oestradiol was elevated to the same high levels by OECP, ovarian and uterine weights were increased in the rat by OECP or DES whereas only the uterus responded in the hamster. This correlated with the ability of the oestrogens to increase significantly the number of large preantral and antral follicles in the intact rat but only the number of follicles with 2-3 layers of granulosa cells in the immature hamster. Qualitative study revealed that DES and OECP increased the number of large preantral follicles in the adult hypophysectomized rat but were ineffective in the adult hamster. It is concluded that for the immature and adult hamster oestrogens do not play a major role in the recruitment of large preantral follicles.

Animals

Development of steroidogenic activity in the ovary of the prepubertal hamster: II. Production of steroids from steroidal precursors and response in vitro to cyclic adenosine monophosphate and luteinizing hormone.

In vitro exposure for 2 h to 250 ng/ml of pregnenolone led to increased production of progesterone and 17 alpha-hydroxyprogesterone (17 alpha-OHP) by hamster ovaries on Days 5, 10 and 15 of age. Similar incubations with 250 ng/ml progesterone or androstenedione caused significant increases in 17 alpha-OHP or testosterone, respectively. When testosterone was added in doses of 32.5, 250 and 500 ng/ml to ovaries on Days 5-30, as early as Day 5 the ovaries aromatized the androgen to estradiol. Day 30 ovaries were the most efficient in the conversion because antral follicles, the principal site for aromatization, were then present. In terms of progesterone production, 400 ng/ml of luteinizing hormone (LH) during 4 h of in vitro incubation stimulated ovaries on Days 5, 10 and 15. Cyclic adenosine 3':5' monophosphate (cAMP) at a dose of 1 mM and 5 mM stimulated progesterone production by Days 5 and 10 ovaries more efficiently than LH. However, Day 15 ovaries produced more progesterone in response to LH compared to cAMP. These experiments establish that the steroidogenic enzymes differentiate at a very early age in the hamster ovary, even before the appearance of gonadotropin receptors. The inability of the early postnatal ovary to produce steroids is apparently attributable to lack of precursors such as cholesterol or cholesterol side chain cleavage enzymes.

Animals

In vivo and in vitro production of progestins by the corpus luteum of pregnancy of the hamster.

Synthesis by hamster corpora lutea (CL) in vitro and serum levels of progesterone (P4), 17 alpha-hydroxyprogesterone (17OHP) and 20 alpha-dihydroprogesterone (20DHP) were assessed for different days of pregnancy (Day 1=day of sperm; Day 16=day of delivery). Highest serum levels of the progestins were observed on Day 14 (P4=33 ng/ml; 17OHP=2 ng/ml; 20DHP=3 ng/ml), followed by precipitous declines on Day 16. The highest in vitro levels of luteal P4 and 17OHP were attained on Days 2-6 (production rates of P4=9-30 ng/mg CL/h; 17OHP=0.6-1.5 ng/mg CL/h), and dropped gradually thereafter. In contrast, the production rate of luteal 20DHP was extremely low on Days 2-8 but abruptly increased on Day 10 and was maintained through Day 14 (4-8 ng/mg CL/h). The in vitro production rates of all 3 progestins dropped abruptly on Day 16. Thus, in the pregnant hamster on the day of parturition (Day 16) there was good agreement between in vivo and in vitro levels of the progestins, contrary to the situation in the rat, where on Day 22 the CL in vitro produce large quantities of P4 and 20DHP while the serum levels are very low (Taya and Greenwald, 1981). Addition of 25 ng/ml of ovine LH to the incubation media containing hamster CL increased production rates of P4 (18-55 ng/mg CL/h) and 17OHP (1.5-2.4 ng/mg CL/h) on Days 2, 4 and 14 of pregnancy but the CL were refractory to this dose of LH from Days 6 to 12. The production rate of luteal 20DHP ws never stimulated by the addition of 25 ng LH. These results indicate that P4 is the principal progestin in the pregnant hamster with 17OHP and 20DHP as minor metabolites. The latter 2 progestins have been measured for the first time in the pregnant hamster and the levels are very low in comparison to the pregnant rat.

Algestone

Autoradiographic analysis of changes in ovarian binding of FSH and HCG during induced follicular atresia in the hamster.

Immediately after hypophysectomy, 30 i.u. PMSG were injected s.c. and 3 days later an antiserum to PMSG was injected i.p. Groups of hamsters were killed at 0,24, 48 and 72 h after PMSG antiserum. The ovaries were prepared for topical autoradiography and the numbers of silver grains in different ovarian compartments were counted. The numbers of binding sites for 125I-labelled FSH in the granulosa cells of the antral follicles dropped sharply to 33, 14 and 5% of that at 0 h at 24, 48 and 72 h respectively. Binding of 125I-labelled hCG to granulosa cells declined more slowly, being 47, 27 and 24%, respectively. Binding of 125I-labelled hCG to thecal and interstitial cells was unaffected. Compared to other models of atresia, the changes in gonadotrophin binding observed in this model occur at an accelerated rate because of the acute deprivation of PMSG.

Animals