The ability of steroid-free bovine follicular fluid to suppress FSH secretion and delay ovulation persists in heifers actively immunized against inhibin

1993 ◽  
Vol 136 (1) ◽  
pp. 137-148 ◽  
Author(s):  
S. C. Wood ◽  
R. G. Glencross ◽  
E. C. L. Bleach ◽  
R. Lovell ◽  
A. J. Beard ◽  
...  

ABSTRACT Previous reports indicate that administration of steroid-free bovine follicular fluid (bFF) to intact heifers suppresses plasma FSH levels and delays the timing of ovulation. In addition, cessation of bFF treatment is associated with a rebound hypersecretion of FSH. To test the hypothesis that these effects of bFF are mediated by inhibin, we have compared the responses to bFF treatment in heifers actively immunized against the N-terminal sequence of inhibin α subunit bIα(1–29)Tyr30-ovalbumin) with those in ovalbumin-immunized controls. Oestrous cycles were synchronized, and inhibin-immune (n = 10) and control (n = 10) heifers were subdivided into two groups which received either 5 ml bFF (n = 5) or 5 ml bovine serum (n = 5) every 4 h for a 60 h period starting 8 h before prostaglandin (PG)-induced luteolysis. Blood was withdrawn every 4 h for 10 days starting 30 h before luteolysis and ovaries were examined daily by ultrasonography. Overall, mean ovulation rate in bIα(1–29)Tyr30-immunized heifers was 44% higher (P < 0·02) than in controls. Inhibin antibody titres tested in bIα(1–29)Tyr30-immunized heifers before (19 ± 3%), during (19 ± 3%) and after (20 ± 3%) bFF treatment did not differ. In the pretreatment period (i.e. mid-luteal phase), plasma FSH levels were 32% (P < 0·05) higher in inhibin-immunized than in control heifers. During administration of bFF to control heifers, plasma FSH was suppressed to a level 40% lower than in serum-treated heifers (P < 0·02). Unexpectedly, bFF suppressed FSH to a similar extent in inhibin-immunized heifers (37% lower than in the serum-treated group; P < 0·025). Similarly, a post-bFF rebound hypersecretion of FSH was observed in both control (P < 0·05) and inhibin-immunized (P < 0·05) heifers, although the FSH rebound lasted about 24 h longer in the latter group (P < 0·001). The timing of the preovulatory LH surge in control (86 ± 7 h post-PG) and immunized (81 ± 6 h post-PG) groups treated with serum was similar as was the timing of the preovulatory rise in plasma oestradiol and the subsequent rise in plasma progesterone. However, bFF treatment delayed (P < 0·001) the preovulatory surges of LH and oestradiol and the subsequent rise in plasma progesterone to a similar extent (> 4 days) in both control and inhibin-immunized groups. Ultrasonography confirmed that bFF delayed the emergence of the wave of dominant ovulatory follicles by 5 days and also showed that inhibin immunization and bFF treatment were both effective in promoting the development of more follicles during the preovulatory period. These results lead us to conclude that a non-steroidal factor(s) in bFF other than inhibin is responsible for the observed delay in ovulation. Like inhibin, this factor acts to suppress pituitary FSH secretion and this could fully account for its ability to delay the onset of preovulatory follicular development and thus delay ovulation. However, the further possibility of a direct inhibitory action of certain bFF components on the ovary cannot be ruled out at this stage. Journal of Endocrinology (1993) 136, 137–148

1999 ◽  
Vol 277 (5) ◽  
pp. E870-E875 ◽  
Author(s):  
Kentaro Nagaoka ◽  
Yasuo Nambo ◽  
Natsuko Nagamine ◽  
Shun-Ichi Nagata ◽  
Yumiko Tanaka ◽  
...  

The relationship between a selective increase in circulating immunoreactive (ir)-inhibin and the time of ovulation was investigated in mares. Concentrations of plasma ir-inhibin were measured every 4 h during the periovulatory period. Inhibin pro-αC, a precursor protein of the inhibin α-subunit, was also measured. The changes in ir-inhibin and inhibin pro-αC in circulation were parallel. Concentrations of both ir-inhibin and inhibin pro-αC in the plasma increased at the same time when ovulatory follicles ruptured, and the peak levels of circulating ir-inhibin and inhibin pro-αC were maintained for 4–8 h. There was no selective increase in plasma concentrations of estradiol-17β during the process of ovulation. These results suggest that the selective increase in ir-inhibin and inhibin pro-αC was caused by the absorption of follicular fluid after the rupture of ovulatory follicles. These results also suggest that the measuring of plasma concentrations of ir-inhibin or inhibin pro-αC in mares might be a useful method for detecting the time of ovulation.


Endocrinology ◽  
2017 ◽  
Vol 159 (2) ◽  
pp. 710-722 ◽  
Author(s):  
Hannah R Bender ◽  
Heidi A Trau ◽  
Diane M Duffy

Abstract Placental growth factor (PGF) is member of the vascular endothelial growth factor (VEGF) family of angiogenesis regulators. VEGFA is an established regulator of ovulation and formation of the corpus luteum. To determine whether PGF also mediates aspects of ovulation and luteinization, macaques received gonadotropins to stimulate multiple follicular development. Ovarian biopsies and whole ovaries were collected before (0 hours) and up to 36 hours after human chorionic gonadotropin (hCG) administration to span the ovulatory interval. PGF and VEGFA were expressed by both granulosa cells and theca cells. In follicular fluid, PGF and VEGFA levels were lowest before hCG. PGF levels remained low until 36 hours after hCG administration, when PGF increased sevenfold to reach peak levels. Follicular fluid VEGFA increased threefold to reach peak levels at 12 hours after hCG, then dropped to intermediate levels. To explore the roles of PGF and VEGFA in ovulation, luteinization, and follicular angiogenesis in vivo, antibodies were injected into the follicular fluid of naturally developed monkey follicles; ovariectomy was performed 48 hours after hCG, with ovulation expected about 40 hours after hCG. Intrafollicular injection of control immunoglobulin G resulted in no retained oocytes, follicle rupture, and structural luteinization, including granulosa cell hypertrophy and capillary formation in the granulosa cell layer. PGF antibody injection resulted in oocyte retention, abnormal rupture, and incomplete luteinization, with limited and disorganized angiogenesis. Injection of a VEGFA antibody resulted in oocyte retention and very limited follicle rupture or structural luteinization. These studies demonstrate that PGF, in addition to VEGFA, is required for ovulation, luteinization, and follicular angiogenesis in primates.


1979 ◽  
Vol 83 (3) ◽  
pp. 355-368 ◽  
Author(s):  
L. V. DEPAOLO ◽  
ANNE N. HIRSHFIELD ◽  
L. D. ANDERSON ◽  
C. A. BARRACLOUGH ◽  
CORNELIA P. CHANNING

In this study, we have examined whether the suppression of raised plasma FSH concentrations at pro-oestrus and/or oestrus by porcine follicular fluid (PFF) affected the development of follicles for ovulation in the next cycle. Adult, 4-day-cyclic rats were injected with PFF or pig serum at various hours of pro-oestrus and oestrus. Plasma FSH levels were suppressed following PFF treatment at any time of pro-oestrus and oestrus. Furthermore, this suppression was always followed by a 'rebound' increase in plasma FSH. In contrast, plasma LH concentrations were unaffected by PFF treatment and neither gonadotrophin was altered by treatment with pig serum. When rats treated with PFF or pig serum were allowed to complete one additional cycle, plasma LH and FSH concentrations at the pro-oestrus and oestrus after treatment were not significantly different among groups regardless of treatment or time of treatment. All ovaries of rats treated with PFF or pig serum on the next pro-oestrus morning before ovulation were histologically similar. Furthermore, all animals ovulated a normal complement of ova at the next oestrus regardless of whether preovulatory, secondary or both increases of plasma FSH had been blocked by PFF treatment during the previous cycle. However, in animals given PFF during the preceding cycle, plasma oestradiol and progesterone concentrations were significantly altered on the morning and afternoon of pro-oestrus respectively. These results suggest that increased plasma FSH concentrations at pro-oestrus and oestrus may not be essential for folliculogenesis and ovulation in the subsequent cycle. Alternatively, the 'rebound' of FSH on day 1 of dioestrus after the suppression of both phases of FSH secretion at pro-oestrus and oestrus may be sufficient to provide ovulatory follicles for the next pro-oestrous day.


1987 ◽  
Vol 115 (1) ◽  
pp. 75-83 ◽  
Author(s):  
A. S. McNeilly ◽  
J. A. Jonassen ◽  
S. M. Rhind

Abstract. The effect of body condition on ovarian follicular development was investigated in Scottish Blackface ewes in high and low body condition. Follicles were dissected from ovaries on days 11 and 12 of the luteal phase and 24 h after prostaglandin-induced luteal regression. Ewes in low body condition had a lower ovulation rate (low: 0.9; high: 1.8 P < 0.05) and lower mean plasma levels of FSH during both the luteal (low: 54; high: 72 μg/l) and follicular (low: 34; high: 43 μg/l) phases of the cycle. Low body condition was associated with a reduced number of large (⩾ 4 mm) follicles in both the luteal and follicular phases, and in low condition a lower proportion of these follicles was oestrogenic and potentially ovulatory as assessed by follicular fluid levels of oestradiol. However, within the different oestrogenic classifications of these large follicles there were no significant differences in the steroidogenic capacity as assessed by the concentrations of either oestradiol or testosterone in follicular fluid, basal and hCG-stimulated testosterone production, thecal 125I hCG binding or basal and testosterone-stimulated oestradiol production by granulosa cells in relation to body condition. These results suggest that body condition influences ovulation rate by altering the concentration of FSH in blood, which in turn affects the number of potentially ovulatory follicles growing beyond 4 mm.


Reproduction ◽  
2000 ◽  
pp. 311-323 ◽  
Author(s):  
JL Hilton ◽  
GE Sarty ◽  
GP Adams ◽  
RA Pierson

The magnetic resonance images and maps of bovine ovaries acquired at defined phases of follicular development and regression were studied to determine whether magnetic resonance image attributes of the follicular antrum reflect the physiological status of dominant and subordinate ovarian follicles. Ovariectomies were performed at day 3 of wave one, day 6 of wave one, day 1 of wave two and at >/= day 17 after ovulation. The timings of ovariectomies were selected to acquire growing, early static, late static and regressing follicles of the first wave and preovulatory follicles of the ovulatory wave. Pre-selection and subordinate follicles were also available for analysis. Serum samples were taken on the day of ovariectomy and follicular fluid samples were taken after imaging. Numerical pixel value and pixel heterogeneity in a spot representing approximately 95% of the follicular antrum were quantified in T(1)- and T(2)-weighted images. T(1) and T(2) relaxation rates (T(1) and T(2)), proton density, apparent diffusion coefficients and their heterogeneities were determined from the computed magnetic resonance maps. The antra of early atretic dominant follicles showed higher T(2)-weighted mean pixel value (P < 0.008) and heterogeneity (P < 0. 01) and lower T(2) heterogeneity (P < 0.008) than growing follicles. Subordinate follicles in the presence of a preovulatory dominant follicle had higher T(1), T(1) heterogeneity, proton density, proton density heterogeneity, and lower mean pixel value in T(1)-weighted images than subordinate follicles of the anovulatory wave (P < 0.04). T(1) relaxation rate heterogeneity and proton density heterogeneity were positively correlated with follicular fluid oestradiol concentration (r = 0.4 and 0.3; P < 0.04). T(2) relaxation rate heterogeneity was positively correlated with follicular fluid progesterone concentration (r = 0.4; P < 0.008). Quantitative differences in magnetic resonance image attributes of the antrum observed among phases of follicular development and regression coincided with changes in the ability of the dominant follicle to produce steroid hormones and ovulate, and thus were indicative of physiological status and follicular health.


1995 ◽  
Vol 145 (3) ◽  
pp. 479-490 ◽  
Author(s):  
B K Campbell ◽  
B M Gordon ◽  
C G Tsonis ◽  
R J Scaramuzzi

Abstract Ewes with ovarian autotransplants received either inhibin antiserum (10 ml i.v. raised in sheep against recombinant 32 kDa human inhibin; n=6) or sheep serum (10 ml i.v.; n=5) on day 3 of the luteal phase with additional daily injections (1 ml i.v.) from 48 h after the initial bolus until day 13. Jugular and ovarian venous blood samples were taken 4-hourly over days 2–13 of the luteal phase. Blood samples were also taken at more frequent intervals (every 10–15 min for 2–3 h) to examine pulsatile secretory responses from the ovary to endogenous and gonadotrophin-releasing hormone-induced (150 ng i.m.) LH pulses on days 4, 6, 8, 10 and 12 of the luteal phase. Plasma FSH levels, ovarian steroid secretion and ovarian follicular development were measured. The ovarian follicle population was estimated daily by real time ultrasound scanning. Immunisation against inhibin resulted in a 3- to 4-fold increase (P<0·001) in plasma FSH levels within 8 h with levels remaining elevated over controls for 6–7 days. Within 24 h of immunisation there was an increase in the number of small ovarian follicles (P<0·05) and by 3 days after treatment immunised ewes had 4–6 large ovarian follicles/ewe with this increase in the total number of large follicles being maintained for the rest of the experimental period (P<0·05). Mean ovarian oestradiol secretion during intensive bleeds was not different from controls 24 h after immunisation, but by 3 days after immunisation it was elevated 4- to 5-fold (P<0·001) over controls with this increase being maintained throughout the experiment. Similar responses to immunisation against inhibin in androstenedione secretion were observed although mean androstenedione secretion was not elevated until 7 days after treatment. In vitro antibody titres in immunised ewes remained elevated but declined steadily (P<0·001) over the experimental period. We conclude that the initial stimulation of follicle development and ovarian steroid secretion following passive immunisation against inhibin can be attributed to increased blood FSH. However, the fact that with time FSH declined but increased follicle development was sustained, despite maintenance of high circulating antibody titres, suggests that on a longer term basis inhibin immunisation may stimulate ovarian function by interfering with the modulation of follicle development by inhibin at an ovarian level. Journal of Endocrinology (1995) 145, 479–490


1986 ◽  
Vol 43 (2) ◽  
pp. 285-292 ◽  
Author(s):  
S. M. Rhind ◽  
B. A. Morris ◽  
Jill Clayton ◽  
J. M. Doney ◽  
R. G. Gunn ◽  
...  

ABSTRACTBorder Leicester × Scottish Blackface (Greyface) ewes of three groups, each comprising 118 animals in a wide range of body condition scores, were mated at a synchronized oestrus in mid October. The ewes were passively immunized against testosterone (group P), actively immunized against androstenedione (group F), or not treated (group C). All ewes were slaughtered at return to service or at 35 to 45 days of pregnancy and ovulation rates and numbers of embryos present were determined. Mean ovulation rates of ewes in group P were higher than in those in group C (P < 0·05) and this difference was evident at most levels of body condition. The absolute increase in ovulation rate, compared with the control group, was similar at all condition scores. Mean ovulation rates of ewes in group F were higher than those in group C (P < 0·001) and the magnitude of the increase was greater in ewes in higher condition scores. The incidence of ova wastage was variable but differences between treatments in mean ovulation rate were generally reflected in mean litter size. The conception rates of immunized ewes were depressed compared with those of control animals, particularly in ewes with a body condition score less than 3·0 at mating. Consequently, there was no improvement in the potential lambing rate of immunized ewes following only one cycle of mating. Circulating antibody titres were not related to conception rate or body condition at mating and were related to ovulation rate only in group F ewes. It is concluded that immunization against steroids, using either passive or active techniques, can improve the reproductive performance of individual ewes but improvement in the performance of the flock as a whole may only be achieved under optimal conditions of nutrition and season.


Reproduction ◽  
2005 ◽  
Vol 129 (4) ◽  
pp. 463-472 ◽  
Author(s):  
Takashi Shimizu ◽  
Izumi Ohshima ◽  
Manabu Ozawa ◽  
Satoko Takahashi ◽  
Atsushi Tajima ◽  
...  

Heat stress inhibits ovarian follicular development in mammalian species. We hypothesized that heat stress inhibits the function of follicular granulosa cells and suppresses follicular development. To test this, immature female rats were injected with pregnant mare serum gonadotropin (PMSG) at 48 h after the start of temperature treatment (control: 25 °C, 50% RH; heat stress: 35 °C, 70% Relative Humidity). The ovaries and granulosa cells of follicles at different developmental stages were analyzed for gonadotropin receptor levels and aromatase activity; estradiol levels were measured in follicular fluid. Before injection, heat stress diminished only the amount of FSH receptor on granulosa cells of antral follicles. During PMSG-stimulated follicular development, heat stress strongly inhibited gonadotropin receptor levels and aromatase activity in granulosa cells, and estradiol levels in the follicular fluid of early antral, antral and preovulatory follicles. To examine apoptosis and mRNA levels of bcl-2 and bax in granulosa cells, follicles harvested 48 h after PMSG injection were cultured in serum-free conditions. Heat-stressed granulosa cells showed a time-dependent increase in apoptosis. The bcl-2 mRNA levels were similar in control and heat-stressed granulosa cells; bax mRNA levels were increased in heat-stressed granulosa cells. According to these results, heat stress inhibits expression of gonadotropin receptors in granulosa cells and attenuates estrogenic activity of growing follicles, granulosa cells of heat-stressed follicles are susceptible to apoptosis, and the bcl2/bax system is not associated with heat-stress-induced apoptosis of granulosa cells. Our study suggests that decreased numbers and function of granulosa cells may cause ovarian dysfunction in domestic animals in summer.


1995 ◽  
Vol 1995 ◽  
pp. 56-56
Author(s):  
M. Khalid ◽  
W. Haresign

Insulin-like growth factor-1 (IGF-1) is one of the potential autocrine/paracrine regulators of ovarian function. Not only do relationships exist between follicular fluid concentrations of IGF-1 and various biochemical markers of follicular differentiation, but IGF-1 has also been shown to stimulate both proliferation and steroidogenesis in ovarian cells in vitro (Adashi et al., 1985). The actions of IGF-1 are thought to be modulated by IGF-binding proteins (IGFBPs). Indeed, follicular growth and atresia in the ewe have been reported to be determined more by changes in IGFBPs than by changes in IGF-1 (Monget et al., 1993). However, in mat particular study, stage of follicular development was determined by follicle size and by microscopic examination of the granulosa cells of individual follicles rather than by biochemical markers of follicle status. The objective of the present study was, therefore, to investigate changes in IGF-1 and IGFBPs levels in follicular fluid and to relate these to the physiological status as determined by steroidogenic content of follicular fluid.


1992 ◽  
Vol 8 (2) ◽  
pp. 109-118 ◽  
Author(s):  
J. Brooks ◽  
W. J. Crow ◽  
J. R. McNeilly ◽  
A. S. McNeilly

ABSTRACT The modulation of FSH secretion at the beginning and middle of the follicular phase of the cycle represents the key event in the growth and selection of the preovulatory follicle. However, the mechanisms that operate within the pituitary gland to control the increased release of FSH and its subsequent inhibition in vivo remain unclear. Treatment of ewes with bovine follicular fluid (bFF) during the luteal phase has been previously shown to suppress the plasma concentrations of FSH and, following cessation of treatment on day 11, a rebound release of FSH occurs on days 12 and 13. When luteal regression is induced on day 12, this hypersecretion of FSH results in an increase in follicle growth and ovulation rate. To investigate the mechanisms involved in the control of FSH secretion, ewes were treated with twice daily s.c. injections of 5 ml bFF on days 3–11 of the oestrous cycle and luteal regression was induced on day 12 with prostaglandin (PG). The treated ewes and their controls were then killed on day 11 (luteal), or 16 or 32h after PG and their pituitaries removed and halved. One half was analysed for gonadotrophin and gonadotrophin-releasing hormone (GnRH) receptor content. Total pituitary RNA was extracted from the other half and subjected to Northern analysis using probes for FSH-β, LH-β and common α subunit. Frequent blood samples were taken and assayed for gonadotrophins. FSH secretion was significantly (P<0.01) reduced during bFF treatment throughout the luteal phase and then significantly (P<0.01) increased after cessation of treatment, with maximum secretion being reached 18– 22h after PG, and then declining towards control values by 32h after PG. A similar pattern of LH secretion was seen after bFF treatment. Pituitary FSH content was significantly (P<0.05) reduced by bFF treatment at all stages of the cycle. No difference in the pituitary LH content was seen. The increase in GnRH receptor content after PG in the controls was delayed in the treated animals. Analysis of pituitary mRNA levels revealed that bFF treatment significantly (P<0.01) reduced FSH-β mRNA levels in the luteal phase. Increased levels of FSH-β, LH-β and α subunit mRNA were seen 16h after PG in the bFF-treated animals, at the time when FSH and LH secretion from the pituitary was near maximum. These results suggest that the rebound release of FSH after treatment with bFF (as a source of inhibin) is related to a rapid increase in FSH-β mRNA, supporting the concept that the rate of FSH release is directly related to the rate of synthesis.


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