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K. Taya and S. Sasamoto

ABSTRACT

Mechanisms responsible for suppression of FSH and LH secretion during lactation were investigated in rats, with special reference to the suckling stimulus and ovarian inhibin. Concentrations of immunoreactive inhibin in the peripheral plasma and bioactive inhibin in ovarian venous plasma were always low on days 3 and 5 of lactation in dams nursing eight pups, whereas values were always high on days 17 and 20 of lactation in dams nursing eight pups and on day 5 of lactation in dams nursing two pups. There was an FSH surge within 48 h after removal of litters on days 3 and 5 of lactation in dams nursing eight pups, whereas plasma concentrations of FSH were unchanged within 48 h by removal of litters on days 17 and 20 of lactation in dams nursing eight pups and on day 5 of lactation in dams nursing two pups. Plasma LH concentrations increased significantly compared with those of control animals within 24 h after removal of the litter on any day of lactation, regardless of the litter size.

Plasma FSH levels increased within 6 h after bilateral or unilateral ovariectomy in lactating rats only on the days when plasma concentrations of inhibin were high before ovariectomy, such as day 17 of lactation in dams nursing eight pups and on day 5 of lactation in dams nursing two pups, whereas the mean concentrations of plasma LH showed no significant increase within 12 h after bilateral ovariectomy in these lactating rats. Treatment with progesterone or oestradiol-17β after unilateral ovariectomy did not inhibit the increase in plasma FSH levels, while the increase in plasma concentrations of FSH after surgery was completely inhibited by injecting inhibin (porcine follicular fluid). Treatment with steroid hormones inhibited the basal levels of LH in unilateral ovariectomized lactating rats. Plasma FSH concentrations increased sharply within 6 h after a single i.v. injection of anti-inhibin serum on days 10, 15 and 20 of lactation in dams nursing eight pups and on day 5 of lactation in dams nursing two pups, whereas only a small but significant increase in concentrations of FSH was noted 6 h after the antiserum treatment on day 5 of lactation in dams nursing eight pups. Concentrations of plasma LH were unchanged by treatment with antiserum in lactating rats throughout lactation.

These findings indicate that the suckling stimulus, rather than ovarian factors, is mainly responsible for the suppression of FSH as well as LH secretion during the first half of lactation in rats nursing eight pups. On the other hand, during the second half of lactation in rats nursing eight pups and throughout lactation in rats nursing two pups, ovarian inhibin plays a primary role in the suppression of FSH secretion, whereas ovarian steroids act to suppress LH secretion.

Journal of Endocrinology (1991) 129, 119–130

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K. Taya and S. Sasamoto

ABSTRACT

Passive immunoneutralization of LHRH by injecting a caprine antiserum to LHRH (LHRH-AS) in lactating rats nursing two pups on day 5 of lactation resulted in an immediate decline in concentrations of LH in the plasma during the 24-h study period, followed by a gradual increase to control levels 30 h later. Concentrations of oestradiol-17β and inhibin activity in ovarian venous plasma also decreased abruptly in LHRH-AS treated animals and recovered to control levels 36 h later. These changes were correlated with changes in concentrations of LH in the plasma. On the other hand, plasma concentrations of FSH increased abruptly in the LHRH-AS treated animals within 3 h after injection, but the concentrations declined gradually to control levels 48 h later. The ability of follicles to ovulate in response to human chorionic gonadotrophin (hCG) began to decrease within 6 h after treatment with LHRH-AS, and further decreased until 18 h after injection of LHRH-AS, when hCG induced ovulation (with two oocytes) in only one of five animals. A gradual increase in ovulation rate to control levels was noted by 36 h after injection of LHRH-AS. These results indicate that Graafian follicles present at the time of LHRH-AS injection had become atretic and that a new set of follicles had then begun to mature. Selective release of FSH could not be induced by injection of LHRH-AS in ovariectomized animals. Treatment with inhibin (porcine follicular fluid) suppressed the selective release of FSH, whereas treatment with oestradiol-17β had no inhibitory effect on the selective release of FSH.

These findings indicate (1) that tonic secretion of LH is an important factor in normal follicular maturation and maintenance in lactating rats, (2) that selective release of FSH after injection of LHRH-AS is attributed to the removal of a negative influence of inhibin from antral follicles during the period of follicular atresia and (3) that a selective surge of FSH is responsible for initiation of new follicular maturation.

J. Endocr. (1988) 118, 455–464

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K. Taya and S. Sasamoto

ABSTRACT

The role of the adrenal gland in the regulation of gonadotrophin and prolactin secretion in the lactating rat was investigated. Changes in secretion of LH, FSH, prolactin, ACTH, β-lipotrophin (β-LPH), inhibin, corticosterone and progesterone after adrenalectomy were examined during the second half of lactation. Follicular maturation was determined by the ability of the follicles to ovulate in response to 10IU human chorionic gonadotrophin (hCG). Adrenalectomy on day 10 of lactation prevented an increase in plasma concentrations of LH and FSH in response to ovariectomy performed at the same time as adrenalectomy, and markedly stimulated secretion of ACTH, β-LPH and prolactin. Adrenalectomy reduced the number of follicles capable of ovulating in response to hCG. Concentrations of inhibin and progesterone in the plasma significantly decreased after adrenalectomy, indicating that development of ovulatory follicles and luteal function had been suppressed. Abolishing the increase in plasma concentrations of LH and inducing a decrease in FSH in the plasma by adrenalectomy therefore prevented maturation of a new set of follicles usually seen during the second half of lactation in rats. The decrease in plasma concentrations of LH also inhibited the ability of the corpus luteum to secrete progesterone, although high concentrations of plasma prolactin were maintained in adrenalectomized lactating rats.

These results indicate that the pituitary-adrenal system is capable of influencing the maintenance of a normal secretion of gonadotrophin and prolactin as well as the maintenance of ovarian function during lactation in the rat.

Journal of Endocrinology (1990) 125, 279—285

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K. Taya and S. Sasamoto

ABSTRACT

To determine whether failure of follicular maturation during the early stages of lactation in rats is due to inadequate LH stimulation, lactating rats nursing eight pups were injected twice daily for 1–3 days (days 2–5 of lactation) with various doses of ovine LH. Follicular maturation was determined by the ability of the follicles to ovulate in response to 10 IU human chorionic gonadotrophin (hCG), endogenous oestradiol-17β and inhibin production. Ovulation was not induced in control animals in response to 10 IU hCG given between days 2 and 5 of lactation. On the other hand, an injection of 10 IU hCG could induce ovulation in LH-treated animals, in which 25 and 50 μg LH per injection were given s.c. from days 2 to 5 of lactation. Concentrations of oestradiol-17β and inhibin activity in ovarian venous plasma increased progressively after the administration of LH, indicating that induced development of ovulatory follicles had occurred. Plasma concentrations of FSH declined in LH-treated animals compared with those in control animals. The decrease in plasma concentrations of FSH was not observed when lactating rats were ovariectomized before the first injection of LH, indicating that ovarian products, probably inhibin, from developing follicles may suppress the secretion of FSH from the pituitary gland. In both LH-treated and control animals, concentrations of prolactin and progesterone remained increased during the period of LH administration. The present results, therefore, suggest that the plasma levels of LH are an important determinant of follicular maturation during lactation in rats.

J. Endocr. (1988) 116, 115–122

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K. Taya and S. Sasamoto

ABSTRACT

The mechanisms responsible for the 24-h difference in the time of the next ovulation after litter removal between early and late lactating rats were investigated.

At 11.00 h on day 5 of lactation, concentrations of oestradiol-17β and inhibin activity in ovarian venous plasma were lower than those on day 17, corresponding to the absence of healthy Graafian follicles. After removal of the litter on day 5 of lactation a small surge of FSH with a steady increase in basal levels of LH occurred to initiate follicular maturation, and ovulation occurred 4 days later. After removal of the litter on day 17 of lactation a surge of FSH was not observed, due to high levels of inhibin activity in ovarian venous plasma, until the time of preovulatory surges of gonadotrophin which occurred 2 days later and resulted in ovulation the next morning. Prolactin concentrations decreased similarly in both groups abruptly after removal of the litter. A decrease in plasma concentrations of progesterone occurred 42 h after removal of the litter on day 5, though it occurred 18 h after removal of the litter on day 17.

These results indicate that the 24-h delay of ovulation after litter removal on day 5 of lactation, as compared with the time of ovulation after litter removal on day 17, is due probably to the absence of healthy antral follicles and high activity of corpora lutea secreting progesterone at the time of litter removal.

J. Endocr. (1987) 113, 271–276

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K. Taya and S. Sasamoto

ABSTRACT

The roles of corticotrophin-releasing factor (CRF) and β-endorphin in the suppression of LH and FSH secretion during lactation were investigated using ovariectomized lactating rats separated from their litters overnight. Within 1 h of returning the pups to their mothers a marked fall in plasma LH concentration and a large increase in plasma prolactin were noted. However, resuckling caused no significant change in plasma concentration of FSH until 12 h after the return of the litter but a significant decline occurred thereafter. Twenty-four hours after removal of the litter, a single i.v. injection of 200 μl anti-LHRH serum caused similar changes in plasma concentrations of LH and FSH observed in nursing rats during suckling. These results suggest that the suckling stimulus itself is responsible for the suppression of LH as well as FSH, via inhibition of the secretion of LHRH. Twenty-four hours after removal of the litter, a single intracerebroventricular (i.c.v.) injection of either 10 μg CRF or β-endorphin resulted in a rapid decrease in plasma LH. Only β-endorphin caused a marked increase in plasma levels of prolactin within 1 h whereas FSH was less affected by either hormone. Repeated i.c.v. administration of 10 μg CRF or β-endorphin at 6-h intervals caused a prolonged inhibition of LH as well as FSH secretion during 48 h, with β-endorphin being less effective than CRF.

These results demonstrate that the suckling stimulus alone suppressed the secretion of both LH and FSH, and suggest that this effect may be mediated by the inhibition of LHRH secretion from the hypothalamus. They also suggest that the suckling-induced inhibition of LHRH may be primarily mediated by endogenous CRF and opioid peptides.

Journal of Endocrinology (1989) 120, 509–515

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A Tohei, G Watanabe and K Taya

The relationship between hypothyroidism and disturbance of the hypothalamo-hypophysial-adrenal axis was investigated using adult male rats. Hypothyroidism was produced by administration of 4-methyl-2-thiouracil (thiouracil) in the drinking water for 2 weeks. Hypothyroidism decreased adrenal weights to 57% of controls and plasma concentrations of corticosterone to 48% of controls. The changes in the weight of adrenals recovered to control levels by administration of thyroxine. The pituitary responsiveness to corticotrophin-releasing hormone (CRH) and arginine vasopressin (AVP) for ACTH release markedly increased in hypothyroid rats as compared with euthyroid rats. In vivo release of CRH and AVP in median eminence significantly increased in hypothyroid rats as compared with euthyroid rats. There were no significant differences in hypothalamic concentrations of CRH and AVP. These results indicate that hypothyroidism causes adrenal dysfunction directly and results in hypersecretion of ACTH mediated by increases in synthesis of CRH and AVP in the hypothalamus.

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G. Watanabe, K. Taya and S. Sasamoto

ABSTRACT

The present study was undertaken to determine whether hypothalamic differentiation is involved in the selective release of FSH during the periovulatory period using adult male rats castrated and implanted with an ovary.

Adult male rats (70–90 days old) were castrated and an ovary obtained from a prepubertal female rat (26 days old) was immediately grafted subcutaneously. Four weeks later, human chorionic gonadotrophin (hCG, 10 i.u.) was injected i.v. into the experimentally manipulated rats to induce ovulatory changes in the grafted ovaries. Another group of similarly prepared rats was injected with 0·9% (w/v) NaCl solution as controls. After injection of hCG, plasma concentrations of FSH increased significantly by 6 h, reached peak values at 12 h and declined to control levels at 36 h. On the other hand, plasma concentrations of LH were reduced by 6 h and decreased further during the next 36 h. An abrupt fall in plasma concentrations of oestradiol-17β occurred within 3 h of the administration of hCG. Histological examination revealed that ovulatory changes and luteinization of follicles were induced in grafted ovaries by 18 h after the injection of hCG. Thirty-six hours after treatment with hCG, a set of newly formed corpora lutea was observed in grafted ovaries and plasma concentrations of progesterone were raised. Treatment with oestradiol-17β did not inhibit the selective release of FSH after the administration of hCG, suggesting that the abrupt decrease in secretion of oestradiol-17β from the grafted ovary is not involved in the occurrence of the FSH surge.

These results indicate that a selective release of FSH can be induced in castrated male rats bearing an ovarian transplant probably due to decreased secretion of inhibin by the luteinized follicles in the grafted ovaries. Sex differentiation of the hypothalamus is not, therefore, involved in the selective surge of FSH.

J. Endocr. (1985) 106, 31–36

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G. Watanabe, K. Taya and S. Sasamoto

ABSTRACT

Plasma and ovarian concentrations of inhibin were determined at 3-h intervals throughout the 4-day oestrous cycle of rats by a radioimmunoassay (RIA) based on a bovine RIA. Plasma concentrations of LH, FSH, progesterone, testosterone, oestradiol-17β, and pituitary contents of FSH and LH were also determined during the cycle. Plasma levels of inhibin showed a marked increase on the morning of oestrus and the afternoon of metoestrus, and a further increase was noted on the morning of pro-oestrus. These increases in plasma levels of inhibin were probably due to the following three events in the ovary, follicular recruitment on the morning of oestrus, selection of follicles on the day of metoestrus, and final maturation of follicles for ovulation on the morning of pro-oestrus with an increase in oestradiol-17β secretion. A striking decrease in inhibin secretion occurred during the process of ovulation after the preovulatory gonadotrophin surge on the afternoon of pro-oestrus. Basal levels of plasma FSH gradually decreased from metoestrus to pro-oestrus as plasma levels of inhibin increased. There was a significant inverse relationship between plasma levels of FSH and inhibin throughout the oestrous cycle (r = −0·51). The present findings suggest that changes in the plasma levels of inhibin during the oestrous cycle provide a precise indicator for follicular recruitment, selection and ovulation, and that changes in concentrations of oestradiol-17β in the plasma are associated with follicular maturation.

Journal of Endocrinology (1990) 126, 151–157

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K. Taya, H. Komura, G. Watanabe and S. Sasamoto

ABSTRACT

Concentrations of inhibin in peripheral blood were determined by radioimmunoassay in pseudopregnant, pregnant and lactating rats and correlated with the status of follicular development, luteal function in the ovaries and placental function. In pseudopregnant rats, plasma concentrations of inhibin remained relatively high from days 2 to 12 of pseudopregnancy and no significant changes occurred throughout pseudopregnancy. In pregnant rats, a marked decline in plasma concentrations of inhibin occurred between days 6 and 14 followed by an abrupt increase on days 20 and 21. In lactating rats, plasma concentrations of inhibin were consistently lower in dams nursing eight pups than in dams nursing two pups. Plasma concentrations of inhibin remained low during the early stages of lactation followed by a significant increase during the late stages in dams nursing eight pups. These changes in inhibin during the period of lactation correlated with those of plasma concentrations of LH, but not with those of oestradiol-17β, testosterone and progesterone. Throughout pseudopregnancy, pregnancy and lactation, on the other hand, plasma concentrations of FSH were within the range of dioestrous levels.

These findings indicate that concentrations of inhibin in peripheral plasma vary with the number of healthy antral follicles and no positive correlation was observed with luteal function during the luteal phase in rats. The results also suggest that inhibin is a more direct index of follicular development than oestradiol-17β in rats.

Journal of Endocrinology (1989) 121, 545–552