Differences in the function and composition of individual ovarian follicles were noted in Booroola Merino ewes which had previously been segregated on at least one ovulation rate record of greater than 5 (FF ewes, N = 15), 3-4 (F+ ewes, N = 18) or less than 3 (++ ewes, N = 18). Follicles in FF and F+ ewes produced oestradiol and reached maturity at a smaller diameter than in ++ ewes. In FF (N = 3), F+ (N = 3) and ++ (N = 3) ewes, the respective mean +/- s.e.m. diameters for the presumptive preovulatory follicles were 3.4 +/- 0.3, 4.1 +/- 0.2 and 6.8 +/- 0.3 mm and in each of these follicles the respective mean +/- s.e.m. numbers of granulosa cells (X 10(6)) were 1.8 +/- 0.3, 2.2 +/- 0.3 and 6.6 +/- 0.3. During a cloprostenol-induced follicular phase, the oestradiol secretion rates from FF ewes with 4.8 +/- 0.4 'oestrogenic' follicles, F+ ewes with 3.2 +/- 0.2 'oestrogenic' follicles and ++ ewes with 1.5 +/- 0.02 'oestrogenic' follicles were not significantly different from one another. Moreover, the mean total numbers of granulosa cells from the 'oestrogenic' follicles from each genotype were identical, namely 5.4 X 10(6) cells. Irrespective of genotype the mean weight of each corpus luteum was inversely correlated to the ovulation rate (R = 0.91, P less than 0.001). Collectively, these findings support the notion that the maturation of greater than or equal to 5 follicles in FF ewes and 3-4 follicles in F+ ewes may each be necessary to provide a follicular-cell mass capable of producing the same quantity of oestradiol as that from 1-2 preovulatory follicles in ++ ewes.
The patterns of growth and atresia of antral follicles including that of the presumptive preovulatory follicle were examined in sheep ovaries for a 24\p=n-\48-h period after the induction of luteolysis with a prostaglandin analogue, cloprostenol or cloprostenol + PMSG. Ewes were ovariectomized at various times after the initiation of the treatments. All follicles \m=ge\1 mm in diameter were dissected from the excised ovaries and the antral fluid and granulosa cells recovered. Individual follicles were classified as healthy or atretic on the basis of the number of granulosa cells recovered and then subclassified as to whether they contained intrafollicular levels of oestradiol that were \ m=ge\ or < than 100 ng/ml. In another series of similarly treated ewes, the ovarian secretion rates of oestradiol and the intrafollicular concentrations of oestradiol in all large antral follicles (\ m=ge\ 5 mm diameter) as well as the levels of progesterone in peripheral plasma were measured at different times after induction of luteolysis. The results showed that a large 'oestrogenic' follicle (\m=ge\5 mm diameter and secreting \ m=ge\ 1ng oestradiol/min) appears around 10 h after the cloprostenol injection and that this presumptive preovulatory follicle emerges before the corpus luteum has ceased to function. Moreover, the presumptive preovulatory ('oestrogenic') follicle appears to develop from the pool of small 'oestrogenic' follicles (1\p=n-\3 mm diameter) after the onset of luteolysis. The emergence of a large 'oestrogenic' follicle is accompanied by a widespread increase in atresia (> 80%) in all other classes of antral follicles (\ m=ge\ 1 mm in diameter). During the first 10 h of cloprostenol-induced luteolysis, PMSG (a) prevented the normal occurrence of atresia in the large follicle population; (b) enhanced oestrogen secretion in a greater proportion of large antral follicles compared to that in control animals; (c) temporarily 'rescued' and/or prevented small antral follicles (1\p=n-\4 mm diameter) from undergoing atresia; but (d) had little, or no, effect on the overall population of antral follicles (\ m=ge\ 1 mm diameter). After 24 h, the atresia-preventing effects of PMSG were no longer discernible and the only obvious difference noted, compared to the controls, was the number of large oestrogen-secreting follicles.
Summary. Injection of steroid-free bovine follicular fluid (bFF; 2 \m=x\5 ml s.c. 12h apart) into anoestrous ewes lowered plasma FSH concentrations by 70% and after 24 h had significantly (P < 0\m=.\01) reduced the number of non-atretic follicles (\ m=ge\ 1 mm diam.) without influencing the total number of follicles (\ m=ge\ 1 mm diam.) compared to untreated controls. Hourly injections of FSH (10 \g=m\g i.v. NIH-FSH-S12) for 24 h did not influence the number of non-atretic follicles but did negate the inhibitory effects of bFF on follicular viability. Hourly injections of FSH (50 \ g=m\ gi.v., NIH-FSH-S12) + bFF treatment for 24 h significantly increased the total number of non-atretic follicles, and particularly the number of medium to large non-atretic follicles (\ m=ge\ 3 mm diam.) compared to the untreated controls (both P < 0\m=.\01). The 10\g=m\g FSH regimen (without bFF) significantly increased aromatase activity in granulosa cells from large ( \ m=ge\ 5 mm diam.; P < 0\m=.\01) but not medium (3\ p=n-\ 4\ m=. \ 5 mm diam.) or small (1\ p=n-\ 2\ m=. \ 5 mm diam.) follicles compared to controls. The 10 \g=m\g FSH + bFF regimen had no effect on granulosa-cell aromatase activity compared to the controls. However, the 50 \g=m\g FSH plus bFF regimen increased the aromatase activity of granulosa cells from large, medium and small non-atretic follicles 2\m=.\6-,8\m=.\3-and \m=ge\11-fold respectively compared to that in the control cells. Ewes (N = 11) that ovulated 2 follicles had significantly higher plasma FSH concentrations from 48 to 24 h and 24 to 0 h before the onset of a cloprostenol-induced follicular phase (both P < 0\m=.\01) than in the ewes (N = 12) that subsequently ovulated one follicle. Hourly FSH treatment (1\m=.\6 \ g =m\ g i.v., NIAMDD-FSH-S15) for 24 h but not for any 6 h intervals between 48 and 24 h or 24 and 0 h before a cloprostenol-induced luteolysis also resulted in significant increases (P < 0\m=.\05) in the number of ewes with 2 ovulations.We conclude that (1) the number of non-atretic antral follicles in sheep ovaries is influenced by plasma FSH concentrations; (2) the level of follicular oestradiol biosynthesis can be enhanced by FSH treatment; and (3) sustained elevations of plasma FSH concentrations for 24 h but not 6 h within 48 h of the onset of luteolysis significantly enhances the ovulation rate in Romney ewes.
A marked difference in both the function and composition of individual ovarian follicles was noted in Booroola X Romney ewes (6-7 years of age) which had previously been segregated on at least one ovulation rate record of 3-4 (F + ewes, N = 21) or less than 3 (++ ewes, N = 21). Follicles in F + ewes produced oestradiol and reached maturity at a smaller diameter than in ++ ewes. In F+ ewes (N = 3), the presumptive preovulatory follicles were 4.4 +/- 0.5 (s.e.m.) mm in diameter and contained 2.1 +/- 0.3 X 10(6) (s.e.m.) granulosa cells, whereas in ++ ewes (N = 3), such follicles were 7.3 +/- 0.3 mm in diameter and contained 6.5 +/- 0.8 X 10(6) cells. During a prostaglandin (PG)-induced follicular phase, the secretion rate of oestradiol from ovaries containing 3 presumptive preovulatory follicles in F + ewes was similar to that from ovaries with only one such follicle in ++ ewes. We suggest that the putative 'gene effect' in F + ewes is manifested during early follicular development and that it may be mediated via an enhanced sensitivity of granulosa cells to pituitary hormones. As a consequence, the development of 3 preovulatory follicles in F + ewes may be necessary to provide a cell mass capable of producing the same quantity of oestradiol as that from one preovulatory follicle in ++ ewes.
Summary. Romney ewes were injected intramuscularly once or twice daily for 3 days with 0, 0\m=.\1,0\m=.\5, 1 or 5 ml of bovine follicular fluid (bFF)
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