Induction of superovulation by immunoneutralization of endogenous inhibin through the increase in the secretion of follicle-stimulating hormone in the cyclic golden hamster

1996 ◽  
Vol 151 (1) ◽  
pp. 65-75 ◽  
Author(s):  
H Kishi ◽  
T Okada ◽  
M Otsuka ◽  
G Watanabe ◽  
K Taya ◽  
...  

Abstract The present study was conducted to study the effect of immunoneutralization against endogenous inhibin on FSH, LH, oestradiol-17β and progesterone secretion and to investigate the effect of removal of endogenous inhibin on subsequent follicular development in the hamster. After treatment with anti-inhibin serum (inhibin-AS) at 1100 h on day 2 of the oestrous cycle (day 1=day of ovulation), a marked increase in plasma levels of FSH and a slight increase in plasma levels of LH were noted and pituitary contents of FSH, but not LH, were also increased. In the group treated with inhibin-AS, superovulation occurred on day 1 of the following cycle. Plasma levels of oestradiol-17β markedly increased with the increase in the number of ovulations induced by human chorionic gonadotrophin (hCG) as compared with those in control animals. In the second cycle, plasma concentrations and pituitary contents of FSH in the animals given 200 μl inhibin-AS still showed high values as compared with those in the animals treated with control serum, although superovulation did not occur on day 1 of the third cycle. Plasma concentrations and pituitary contents of LH in the hamster given 200 μl inhibin-AS tended to decrease as compared with those in control animals during the second cycle. Plasma concentrations of oestradiol-17β in the animals treated with 200 μl inhibin-AS changed in a similar way to controls. A marked increase in plasma concentrations of progesterone was noted on days 1 and 2 of the second cycle in the group receiving inhibin-AS. The twice daily injection of 1 IU hCG during the second cycle to the animals given 200 μl inhibin-AS induced superovulation on day 1 of the third cycle. These results indicate that circulating inhibin may be an important indicator of the number of developing follicles and may maintain the species-specific number of developing follicles through suppression of FSH secretion in the cyclic hamster. They also suggest that high levels of inhibin slightly suppress plasma levels of LH, indicating that plasma LH may also regulate follicular development in the cyclic hamster. Journal of Endocrinology (1996) 151, 65–75

1995 ◽  
Vol 146 (1) ◽  
pp. 169-176 ◽  
Author(s):  
H Kishi ◽  
K Taya ◽  
G Watanabe ◽  
S Sasamoto

Abstract Plasma and ovarian levels of inhibin were determined by a radioimmunoassay (RIA) at 3-h intervals throughout the 4-day oestrous cycle of hamsters. Plasma concentrations of FSH, LH, progesterone, testosterone and oestradiol-17β were also determined by RIAs. In addition, hamsters were injected at various times with human chorionic gonadotrophin (hCG) to determine the follicular development. The changes in plasma concentrations of FSH after injection of antisera to oestradiol-17β (oestradiol-AS) and inhibin (inhibin-AS) on the morning of day 2 (day 1=day of ovulation) were also determined. Plasma concentrations of inhibin showed a marked increase on the afternoon of day 1, remained at plateau levels until the morning of day 4, then increased abruptly on the afternoon of day 4 when preovulatory LH and FSH surges were initiated. A marked decrease in plasma concentrations of inhibin occurred during the process of ovulation after the preovulatory gonadotrophin surges. An inverse relationship between plasma levels of FSH and inhibin was observed when the secondary surge of FSH was in progress during the periovulatory period. Plasma concentrations of oestradiol-17β showed three increase phases and these changes differed from those of inhibin. Changes in plasma concentrations of oestradiol-17β correlated well with the maturation and regression of large antral follicles. Follicles capable of ovulating following hCG administration were first noted at 2300 h on day 1. The number of follicles capable of ovulating reached a maximum on the morning of day 3 (24·8± 0·6), and decreased by 0500 h on day 4 (15·0 ± 1·1), corresponding to the number of normal spontaneous ovulations. Plasma concentrations of FSH were dramatically increased within 6 h after inhibin-AS, though no increase in FSH levels was observed after oestradiol-AS. These findings suggest that changes in the plasma levels of inhibin during the oestrous cycle provide a precise indicator of follicular recruitment, and that the changes in plasma concentrations of oestradiol-17β are associated with follicular maturation. These findings also suggest that inhibin may play a major role in the inhibition of FSH secretion during the oestrous cycle of the hamster. Journal of Endocrinology (1995) 146, 169–176


1991 ◽  
Vol 130 (2) ◽  
pp. 313-320 ◽  
Author(s):  
P. C. Wynn ◽  
M. C. Stuart ◽  
A. L. C. Wallace ◽  
A. C. Kirby ◽  
E. F. Annison

ABSTRACT The effect of daily administration of ovine GH for a period of 4 weeks on somatomedin-C biological activity in plasma was investigated in mature Merino sheep fed a maintenance energy intake (low plane; LP) or 1·6 times this amount (high plane; HP). The GH treatment resulted in a significant (P < 0·05) increase in plasma GH levels in blood samples collected 23·5 h after each daily injection in both LP and HP groups. Plasma concentrations of somatomedin-C activity and insulin were significantly stimulated to a maximum level by the third GH injection and remained at this level for 7 days. Subsequently, circulating levels of both hormones fell to 40–50% of the peak response to GH and returned to basal levels within 48 h of the cessation of GH injections. In the HP group the response of plasma insulin and somatomedin-C activity to GH injection was greater than in the LP group. Journal of Endocrinology (1991) 130, 313–320


1976 ◽  
Vol 71 (1) ◽  
pp. 51-58 ◽  
Author(s):  
R. J. ETCHES ◽  
F. J. CUNNINGHAM

SUMMARY The existence of a circadian rhythm in the sensitivity of the hypothalamus of the laying hen to stimulation by progesterone was investigated by injecting 0·5 mg progesterone subcutaneously during the proposed period of maximum insensitivity. Following this treatment increases in plasma concentrations of both LH and progesterone were observed which were comparable to the spontaneous preovulatory rises in the plasma levels of the hormones. The ability of either progesterone or luteinizing hormone releasing hormone (LH-RH) to induce premature ovulation varied according to the stage of follicular development. Neither hormone was more than 28% effective when injected within 6·5 h of the previous ovulation, whereas both hormones were 100% effective approximately 27 h after the terminal ovulation of a clutch sequence. Failure to ovulate in response to LH-RH given 6·5 h after ovulation was associated with a lack of progesterone secretion. Both LH and progesterone were secreted when ovulation was induced by injections of either LH-RH or progesterone, and LH was secreted in response to progesterone given 6·5 h after ovulation. These results demonstrate that progesterone stimulates the secretion of LH and LH stimulates the secretion of progesterone. The precise physiological role of these two hormones, however, was not established.


1980 ◽  
Vol 87 (3) ◽  
pp. 393-400 ◽  
Author(s):  
KAZUYOSHI TAYA ◽  
SHUJI SASAMOTO

In order to elucidate the mechanism of the resumption of follicular activity and ovulation in rats, levels of FSH, LH and prolactin in plasma and pituitary gland and ovarian follicular development were quantified after removal of the litter on day 3 of lactation (day of parturition = day 0 of lactation). Such removal resulted in ovulation of 13 oocytes 4 days later, a number comparable with that found in normal cyclic rats. Plasma levels of prolactin were high during lactation but markedly decreased after removal of the litter. Although plasma concentrations of FSH and LH did not change during days 3–7 of lactation, there was an FSH surge between 24 and 30 h after removal of the litter. Plasma concentrations of LH also increased slightly but significantly by 24 h after removal of the litter and this value persisted during the following 2 days. Surges of FSH, LH and prolactin occurred at 17.00 h 3 days after pups were removed. Removal of the litter did not increase pituitary contents of FSH, LH and prolactin and a marked reduction in pituitary levels of FSH and LH, but not of prolactin, occurred at 17 00 h 3 days after removal of the litter. A quantitative study of follicular development indicated that follicles larger than 401 μm in diameter were absent during days 3–7 of lactation. However, the number and size of antral follicles increased by 30 h after removal of the litter, probably due to the increases in plasma levels of FSH and LH, and follicles larger than 601 μm in diameter appeared 3 days after the young were removed. Although ovulation could not be induced by human chorionic gonadotrophin from days 3 to 5 of lactation, its administration 30 h after removal of the litter produced ovulation in all rats by the following morning. These results indicated that a moderate increase in FSH, although below the amounts released at the preovulatory surge, together with basal levels of LH which were within the range observed on the day of dioestrus during the normal cycle were responsible for the initiation of follicular maturation after removal of the litter.


1993 ◽  
Vol 136 (1) ◽  
pp. 35-41 ◽  
Author(s):  
H. Kaneko ◽  
Y. Nakanishi ◽  
K. Taya ◽  
H. Kishi ◽  
G. Watanabe ◽  
...  

ABSTRACT To investigate the physiological importance of inhibin in the regulation of FSH secretion in cows, seven cyclic cows were treated with an inhibin antiserum raised against bovine 32 kDa inhibin in a castrated goat. The same animals treated with a castrated goat serum (control serum) served as controls. On day 12 of the oestrous cycle (day 0 = day of oestrus), four of seven cows were injected with 100 ml inhibin antiserum first, and the remaining three cows with 100 ml control serum first. Twelve days after the second oestrus following the first serum injection (42–46 days after the first serum injection), the former four cows were injected with control serum and the latter three with inhibin antiserum. Follicular development after the injections of control serum or inhibin antiserum was assessed by daily ultrasonographic examination. Treatment with inhibin antiserum resulted in a marked increase (P < 0·01) in plasma concentrations of FSH and oestradiol-17β but not LH or progesterone, compared with those after treatment with control serum. Plasma concentrations of FSH increased significantly (P < 0·01) at 8 h after injection of antiinhibin serum when compared with the control value. Concentrations of FSH in the plasma remained high for 72 h, then declined to the control level by 84 h, concomitant with an abrupt decrease in the titre of free inhibin antibody in the plasma. High concentrations of oestradiol-17β were observed between 36 and 96 h after treatment. Treatment with inhibin antiserum markedly increased the number of small (≥ 4 < 7 mm in diameter), medium (≥ 7 < 10 mm) and large (≥ 10 mm) follicles by 48, 72 and 96 h after treatment when compared with the value before treatment. The number of large follicles returned to the pretreatment value at 168 h, whereas the number of small and medium follicles remained increased. The present results provide strong evidence that inhibin is an important factor in the inhibitory regulation of FSH secretion during the mid-luteal phase of cows, and demonstrate that an increase in endogenous FSH secretion after immunoneutralization of circulating inhibin stimulates the rapid growth of a large number of follicles. Journal of Endocrinology (1993) 136, 35–41


1999 ◽  
Vol 82 (11) ◽  
pp. 1428-1432 ◽  
Author(s):  
Cheryl Scott ◽  
Francesco Salerno ◽  
Elettra Lorenzano ◽  
Werner Müller-Esterl ◽  
Angelo Agostoni ◽  
...  

SummaryLittle is known about the regulation of high-molecular-weight-kininogen (HK) and low-molecular-weight-kininogen (LK) or the relationship of each to the degree of liver function impairment in patients with cirrhosis. In this study, we evaluated HK and LK quantitatively by a recently described particle concentration fluorescence immunoassay (PCFIA) and qualitatively by SDS PAGE and immunoblotting analyses in plasma from 33 patients with cirrhosis presenting various degrees of impairment of liver function. Thirty-three healthy subjects served as normal controls. Patients with cirrhosis had significantly lower plasma levels of HK (median 49 μg/ml [range 22-99 μg/ml]) and LK (58 μg/ml [15-100 μg/ml]) than normal subjects (HK 83 μg/ml [65-115 μg/ml]; LK 80 μg/ml [45-120 μg/ml]) (p < 0.0001). The plasma concentrations of HK and LK were directly related to plasma levels of cholinesterase (P < 0.0001) and albumin (P < 0.0001 and P < 0.001) and inversely to the Child-Pugh score (P < 0.0001) and to prothrombin time ratio (P < 0.0001) (reflecting the clinical and laboratory abnormalities in liver disease). Similar to normal individuals, in patients with cirrhosis, plasma HK and LK levels paralleled one another, suggesting that a coordinate regulation of those proteins persists in liver disease. SDS PAGE and immunoblotting analyses of kininogens in cirrhotic plasma showed a pattern similar to that observed in normal controls for LK (a single band at 66 kDa) with some lower molecular weight forms noted in cirrhotic plasma. A slight increase of cleavage of HK (a major band at 130 kDa and a faint but increased band at 107 kDa) was evident. The increased cleavage of HK was confirmed by the lower cleaved kininogen index (CKI), as compared to normal controls. These data suggest a defect in hepatic synthesis as well as increased destructive cleavage of both kininogens in plasma from patients with cirrhosis. The decrease of important regulatory proteins like kininogens may contribute to the imbalance in coagulation and fibrinolytic systems, which frequently occurs in cirrhotic patients.


1987 ◽  
Vol 58 (03) ◽  
pp. 850-852 ◽  
Author(s):  
M B McCrohan ◽  
S W Huang ◽  
J W Sleasman ◽  
P A Klein ◽  
K J Kao

SummaryThe use of plasma thrombospondin (TSP) concentration was investigated as an indicator of intravascular platelet activation. Patients (n = 20) with diseases that have known vasculitis were included in the study. The range and the mean of plasma TSP concentrations of patients with vasculitis were 117 ng/ml to 6500 ng/ml and 791±1412 ng/ml (mean ± SD); the range and the mean of plasma TSP concentrations of control individuals (n = 33) were 13 ng/ml to 137 ng/ml and 59±29 ng/ml. When plasma TSP concentrations were correlated with plasma concentrations of another platelet activation marker, β-thromboglobulin (P-TG), it was found that the TSP concentration inei eased exponentially as the plasma β-TG level rose. A positive correlation between plasma levels of plasma TSP and serum fibrin degradation products was also observed. The results suggest that platelets are the primary source of plasma TSP in patients with various vasculitis and that plasma TSP can be a better indicator than β-TG to assess intravascular platelet activation due to its longer circulation half life.


1996 ◽  
Vol 76 (06) ◽  
pp. 1090-1095 ◽  
Author(s):  
C Ravanat ◽  
M Freund ◽  
S Schuhler ◽  
P Grunert ◽  
L Meyer ◽  
...  

SummaryThe purpose of this study was to develop specific and sensitive immunoassays to detect early indices of hypercoagulability in the rat. Rat platelet factor 4 (rPF4) and rat fibrinopeptide A (rFPA) assays, tools for the detection of activation of platelets and coagulation respectively, were designed using antibodies raised against purified rPF4 and against synthetic rFPA. The relevance of these new assays and of the commercially available ELISA kit for thrombin-antithrombin III (TAT) complexes was demonstrated in a rat model of a prethrombotic state induced by intravenous infusion of varying doses of thromboplastin (90 to 2400 μl/kg/h). In this model, the immunoassays allowed simultaneous detection of low levels of rFPA and rPF4 which were correlated with fibrinogen and platelet consumption and TAT generation and further proved to be of higher sensitivity than the classical methods of platelet count or measurement of fibrinogen levels. Plasma concentrations of rFPA, rPF4 and TAT were dependent on infusion time and thromboplastin dose, while hirudin (1 mg/kg) prevented their appearance. Thus the new specific immunoassays for rPF4 and rFPA and the commercial human TAT assay represent useful tools for pathophysiological studies or the screening of antithrombotic drugs in rats.


1996 ◽  
Vol 76 (03) ◽  
pp. 328-332 ◽  
Author(s):  
Bernd Jilma ◽  
Peter Fasching ◽  
Christine Ruthner ◽  
Anna Rumplmayr ◽  
Sabine Ruzicka ◽  
...  

SummaryBased on findings that showed increased P-selectin expression on platelets and on choroidal microvessels of patients with insulin dependent diabetes mellitus (IDDM), we hypothesized that also plasma concentrations of circulating (c)P-selectin would be increased in these patients.The aim of this study was to compare the plasma levels of cP-selec-tin between non-smoking patients with IDDM, treated with an intensified insulin therapy, and healthy controls. The study design was prospective, cross-sectional and analyst-blinded. Subjects were matched individually for sex, age and body mass index. Plasma levels of cP-selectin and of von Willebrand antigen (vWF-Ag) were determined by enzyme linked immunoassays.Forty-two pairs were available for intergroup comparison. Median plasma concentrations of cP-selectin in patients with IDDM (285 ng/ml; interquartile range: 233-372) were on average 21% higher than those of controls (236 ng/ml; interquartile range: 175-296; p = 0.004). Also, median plasma levels of vWF-Ag were 10% higher in patients (96 U/dl; interquartile range: 82-127) than controls (87 U/dl; interquartile range: 70-104; p = 0.025). There was no correlation between plasma concentrations of cP-selectin and vWF-Ag levels in either group (p ώ0.05).In conclusion, our results of increased cP-selectin levels are in line with increased P-selectin expression on platelets and on choroidal microvessels found in patients with IDDM. In view of the currently developed small molecule inhibitors of cell adhesion molecules, these independent observations together may provide a sound rationale to select P-selectin as a target for treating or preventing IDDM-associated micro- or macrovascular complications.


1972 ◽  
Vol 69 (2) ◽  
pp. 267-280 ◽  
Author(s):  
Richard D. Peppler

ABSTRACT Intact 5-day cycling rats were killed between 8–10 a. m. on each day of the oestrous cycle; experimental rats were unilaterally ovariectomized (ULO) at 9 a. m. on day 1 (oestrus) and killed between 8 and 10 a. m. on days 2, 3, 4 or 1 of the subsequent cycle. Pituitary and plasma concentrations of FSH and LH were measured in both groups of rats. Pituitary FSH concentration was measured by the Steelman-Pohley method with slight modification; plasma FSH by the Igarashi-McCann assay and pituitary and plasma LH concentration by the OAAD method. In intact rats, pituitary FSH values remained constant for the first three days of the cycle, increased on day 4 and reverted to early cycle values by day 5. Plasma FSH increased between days 2 and 3 and days 5 and 1. Pituitary LH concentration remained the same for days 1 and 2; increased two-fold on days 3 and 4, and increased further by day 5. Plasma LH increased between days 2 and 3; other differences between successive cycle days were not apparent. Following ULO on day 1, pituitary FSH increased steadily, but not significantly, for the remaining cycle. Plasma FSH did not change from day 2 through day 1 of the subsequent cycle. Pituitary LH remained low on day 2, increased sharply by day 3 and decreased (50 %) by day 4. Plasma LH also increased between days 2 and 3. Other differences between successive days following unilateral ovariectomy on day 1 were not apparent. Correlation of gonadotrophin activity with follicular development suggests that the mechanism of compensatory ovulation in the rat may be one of an increase in time of exposure to a constant gonadotrophic level for the duration of the oestrous cycle rather than to increased levels of the gonadotrophin.


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