Plasma LH,FSH, testosterone,and age at puberty in ram lambs actively immunized against an inhibin alpha-subunit peptide

Active immunization against inhibin has been shown to advance puberty and increase ovulation rate in ewe lambs; but in ram lambs, effects on puberty and sperm production are equivocal. The objective of the present study was to determine whether active immunization against an inhibin alpha-subunit peptide advances the onset of puberty in ram lambs. St. Croix hair sheep ram lambs were assigned to inhibin-immunized (n = 7) and control (n = 8) treatment groups. Lambs in the inhibin-immunized group were immunized against a synthetic peptide-carrier protein conjugate, alpha-(1-25)-human alpha-globulin (halpha-G), and control lambs were immunized against halpha-G. Lambs were immunized at 3, 7, 13, 19, 25, 31, and 37 weeks of age. On the day of immunization a blood sample was collected and lambs were weighed. Another blood sample was collected 1 week following each immunization. At 20 weeks of age additional blood samples were collected at 20 min intervals for 8h. Beginning at 20 weeks of age and at weekly intervals thereafter, scrotal circumference (SC) was measured and semen was collected using electroejaculation. A subsequent ejaculate was collected 1 week following onset of puberty, which was defined as the week of age when an ejaculate first contained > or =50 x 10(6) sperm cells. In control lambs, plasma alpha-(1-25)-antibody (Ab) was nondetectable. In inhibin-immunized lambs, alpha-(1-25)-Ab titer increased from 7 to 25 weeks of age and then plateaued at a level that varied (P<0.001) among animals. Body weight and SC of control and inhibin-immunized lambs were similar at the onset of puberty. At pubertal onset inhibin-immunized lambs were older than control lambs (31.9+/-0.5 vs. 29.5+/-0.7 weeks of age, P<0.05). Plasma FSH concentrations were similar in control and inhibin-immunized lambs from 3 to 38 weeks of age. Plasma LH levels were lower (P<0.01) in inhibin-immunized than control lambs. During the 8-h blood sampling period at 20 weeks of age, LH and testosterone concentrations were lower (P<0.05) in inhibin-immunized than control ram lambs, and the LH pulse frequency was similar in the two groups of animals. The decreased LH secretion is consistent with the immunoneutralization of a putative inhibin alpha-subunit-related peptide that stimulates LH secretion in ram lambs. Present findings show that active immunization against an inhibin alpha-peptide delays rather than advances puberty in ram lambs.     

Reproductive characteristics of lambs actively immunized early in life with inhibin-enriched preparations from follicular fluid of cows

Active immunization of Merino and crossbred ewe lambs early in life with a partly purified inhibin derived from bovine follicular fluid (bFF) advanced their puberty. This occurred whether the lambs were immunized from 3 weeks of age or from 9 weeks of age or just 3 times between 3 and 9 weeks. However, the effect was more obvious with multiple injections starting at 3 weeks of age. The ovulation rate of the immunized lambs was significantly higher than that of the control ewes. When lambs were subjected to multiple immunizations the increased ovulation rate persisted in Merinos for at least 1 year after immunization ceased. The lambs injected 3 times only (at 3, 6 and 9 weeks of age) did not show any increase in ovulation rate. Plasma concentrations of FSH and LH measured in blood samples taken 1 week after immunization were not significantly different between the treatment groups. Daily sperm output in the urine and testis diameter were measured in ram lambs from the same breeds and flocks as the ewe lambs. Immunization with the inhibin preparation from 3 weeks of age resulted in a significant increase in daily sperm output (4255 +/- 701 vs 2344 +/- 344 x 10(6), P less than 0.01) and testis diameter (5.21 +/- 0.3 vs 4.33 +/- 0.2 cm, P less than 0.05) in Merino rams examined in the non-breeding season when the rams were aged 23 months. Although the same trend was seen in the following year the differences were not significant. Immunization from 9 weeks of age had no effect. A similar increase in daily sperm output was seen (at 20 and 25 months of age) in crossbred rams immunized from 3 weeks of age but the difference was not significant. Plasma concentrations of FSH (but not LH) in samples from rams immunized from 3 weeks of age were significantly higher than those of control rams at 7 and 60 weeks of age in Merino rams and at 23 and 31 weeks of age in crossbred rams. Plasma FSH concentrations at 21 and 26 weeks of age in the Merino rams immunized from 3 weeks of age were higher than those of control lambs, and these increases preceded a significant (P less than 0.05) increase in testicular diameter at 30 weeks of age (4.99 +/- 0.2 vs 4.37 +/- 0.2 cm). These results suggest that active immunization early in life with an inhibin-enriched fraction from bFF advances puberty in ewe lambs.(ABSTRACT TRUNCATED AT 400 WORDS)     

Effect of immunizing prepuberal lambs of low and high ovulation rate genotypes with inhibin partially purified from bovine follicular fluid

Active immunization of prepuberal lambs with a partially purified inhibin preparation, isolated from bovine follicular fluid, increased the ovulation rate. In ewe lambs of a low fecundity breed (Suffolk x Galway), the ovulation rate rose from 1.15 to 1.95 (P<0.05) compared with that of the controls. An ovulation rate of 3.38 was recorded for immunized ewe lambs of a high fecundity breed (Finn x Dorset Horn), while the rate for mature ewes from the same flock was 2.29. Immunization did not affect the time of onset of puberty or estrous cycle length. Following immunization, antibodies were produced that bound to a pure preparation of 68kDa bovine inhibin. This report demonstrates the production of antibody to a 68kDa preparation of inhibin following active immunization of sheep using a partially purified preparation. It was concluded that the increased ovulation rate was due to the production of antibodies to inhibin, which may have reduced its negative feedback effect of FSH secretion.     

Relationship between follicle growth and circulating gonadotrophin levels during postnatal development of sheep

This study investigates the number and size of ovarian antral follicles in relation to plasma follicle stimulating hormones (FSH) and luteinizing hormone (LH) concentrations from birth to 26 weeks of age in ewe lambs of the Ouled Djellel breed, a non-seasonal breed of sheep. Plasma was collected from 10 ewe lambs at 14 sampling times (Week 0, i.e. <24h, Week 1 and every two weeks from Week 4 to Week 26, inclusive). At each of these stages, four ewe lambs were slaughtered, the ovaries recovered and weighed, and the number and size of the follicles determined from histological examination. The pattern for plasma FSH showed a peak at Week 10, a smaller peak at Week 18 and a very small peak at Week 24. The pattern for LH was similar until Week 24 when the largest peak occurred. Paired ovarian weight increased rapidly from birth to four weeks and then more slowly to 10 weeks, followed by a decline at 12 weeks and a gradual increase from 14 to 24 weeks of age. The number and total diameter of follicles > or =3 mm in diameter showed similar patterns of development--rising gradually from birth to Week 14, falling to Week 16 and then rising more rapidly to a peak at Week 24. Maximum follicle diameter declined from birth to Week 1, then rose rapidly to Week 4, followed by a more gradual rise to Week 14 and, thereafter, a more rapid increase to a peak of 7.23+/-0.16 mm at 24 weeks old. The number of follicles (<3 mm diameter) increased rapidly from birth to Week 10 and then declined to values similar to those at Weeks 1 and 4. First behavioural oestrus was observed at Week 24 and a corpus luteum was present on the ovary of one lamb at Week 24 and two lambs at Week 26. It was concluded that two or three peaks in plasma FSH and LH levels precede puberty and first ovulation in Ouled Djellel ewe lambs, and first ovulation occurred at 24-26 weeks of age. The increase in follicle number and size generally reflected the pattern of plasma FSH and LH levels.     

Identifying infertile homozygous Inverdale (FecXI) ewe lambs on the basis of genotype differences in reproductive hormone concentrations

Introduction of the Inverdale prolificacy gene (FecX^I) could markedly improve reproductive efficiency in commercial flocks, but as homozygous carrier Inverdale ewes are infertile, it is imperative that these animals are identified at an early age and excluded from breeding stock. As the ovaries of homozygous carrier ewes are nonfunctional, there are wide differences in reproductive hormone levels between these and other Inverdale genotypes. This study assesses the accuracy of using hormone concentrations alone, to identify infertile homozygous ewe lambs. Ewe lambs were blood sampled at 2, 5 and/or 8 months of age, and plasma analyzed for follicle-stimulating hormone (FSH), luteinizing hormone (LH) and inhibin content. These animals were either the offspring of both known carrier rams and known carrier ewes, and therefore would be either homozygous (II) or heterozygous (I +) for the Inverdale gene (group 1, N = 122), or had one parent that was a carrier and therefore would be either heterozygous or noncarriers (+ +) of the gene (group 2, N = 32). Animals were designated as either II or I + / + + on the basis of their plasma hormone concentrations. Inverdale genotype was also assigned from laparoscopic observation of the ovaries at each of these occasions. Definitive assignment of genotype was made at laparoscopy as adults during the breeding season. On the basis of laparoscopy as adults, 62 (51%) lambs in group 1 were identified as homozygous and 60 (49%) as heterozygous. At all three ages, both mean FSH and mean LH concentrations were significantly higher in II than in I + lambs. Mean inhibin concentrations were significantly lower in II lambs at 8 months, but did not differ significantly between genotypes at 2 or 5 months of age. The use of discriminant analysis techniques to segregate individual animals in group 1 on the basis of their plasma FSH and LH concentrations, correctly identified Inverdale genotype in 50/52 (96%) lambs at 2 months, 75/79 (95%) at 5 months and 118/122 (97%) at 8 months of age. Discriminant analysis was equally effective for segregating II ewe lambs (group 1) from fertile ewe lambs of I + and + + genotype (group 2, 97% correct at 5 months and 98% at 8 months). At no stage did inclusion of inhibin concentrations into the discriminant function alter the number of homozygous ewes misclassified. This demonstrates that infertile homozygous ewe lambs can accurately be distinguished from their fertile flockmates by using plasma concentrations of gonadotrophins alone, and that this can be achieved from as early as 2 months of age.     

Effect of inhibin immunisation using different synthetic peptide fragments of the bovine αc-subunit on plasma anti-inhibin titres,plasma FSH concentrations and the incidence of multiple ovulation in heifers

In an effort to improve the effectiveness of inhibin immunisation in promoting multiple ovulation in cattle and to clarify the mechanism(s) involved, heifers (n = 5 per group) were immunised against ovalbumin conjugates of different synthetic peptide sequences of the α_c-subunit of bovine inhibin (bIα) selected using antigenic prediction methods. Plasma inhibin antibody titre (percentage binding of ¹²⁵I-labelled M_r 32 000 native bovine inhibin), plasma follicle-stimulating hormone (FSH) concentration and ovulatory response (number of corpora lutea observed by transrectal ovarian ultrasonography) were recorded over a 16 week period.Heifers immunised against the bIα1–29 and 63–72 peptides (alone or in combination), had relatively high anti-inhibin titres (over 7.5% binding) and showed a significantly (P < 0.05) increased incidence of multiple ovulations (18–65%) compared with ovalbumin-immunised controls. However, immunisation against the 1–16 and 108–123 peptides was relatively ineffective in generating antibodies reactive with native inhibin (less than 7.5% binding) and gave little or no increase in incidence of multiple ovulations (0–10%).Analysis of results for all 33 heifers revealed a significant linear relationship between mean inhibin antibody titre and mean plasma FSH concentration (r = 0.42; P < 0.02) and between mean inhibin antibody titre and incidence of multiple ovulation (r = 0.89; P < 0.0001). A significant quadratic relationship existed between mean inhibin antibody titre and the mean number of ovulations per cycle (r = 0.88; P < 0.0001). However, partial correlation analysis showed a highly significant association between anti-inhibin titre and ovulatory response which was independent of changes in mean plasma FSH concentrations.These results extend previous studies involving inhibin peptide-immunised cattle by showing that the magnitude of the ovulatory response is directly related to the prevailing titre of antibodies reactive with native inhibin. However, they do not support the hypothesis that the ovulatory response is mediated solely by a rise in FSH secretion.     

Inhibin B,follicle stimulating hormone,luteinizing hormone and testosterone during childhood and puberty in males: changes in serum concentrations in relation to age and stage of puberty

Inhibin B is a gonadal dimeric polypeptide hormone that regulates synthesis and secretion of follicle stimulating hormone (FSH) in a negative feedback loop. The aim of the present study was to determine changes in serum inhibin B, gonadotropins and testosterone concentrations during childhood and puberty in males. We studied the relationship between circulating inhibin B, gonadotropins and testosterone in serum of healthy boys during the first two years of life and then in pubertal development. Using a recently developed two-side enzyme-linked immunosorbent assay (ELISA), inhibin B levels were measured in the serum of 78 healthy boys divided into eleven age groups from birth to the end of pubertal development. In addition, serum levels of gonadotropins and testosterone were measured. Serum inhibin B, gonadotropins and testosterone increased during the first months of postnatal life. A peak in serum inhibin B and gonadotropins concentrations was observed around 3-4 months of age. There was a significant positive correlation between serum inhibin B and gonadotropins and testosterone levels during the first 2 years of life. After this early increase, serum inhibin B, gonadotropins and testosterone levels decreased significantly and remained low until puberty followed by an increase beginning with the onset of puberty. Serum levels of inhibin B reached a peak at stage G3 of puberty. Around midpuberty, inhibin B lost its positive correlation with luteinizing hormone (LH) and testosterone from early puberty, and developed a strong negative correlation with FSH, which persisted into adulthood. We conclude that inhibin B plays a key role in the regulation of the hypothalamic-pituitary-gonadal hormonal axis during male childhood and pubertal development. Inhibin B is a direct marker of the presence and function of Sertoli cells and appears to reflect testicular function in boys.     

Pituitary response to LHRH, LH pulsatility and plasma melatonin and prolactin changes in ewe lambs treated with melatonin implants to delay puberty

Prepubertal ewe lambs were treated with empty or filled melatonin implants. The implants were placed s.c. at birth and pituitary responsiveness to various doses of LHRH, LH/FSH pulsatility and prolactin and melatonin secretion were examined at 10, 19, 28, 36 and 45 weeks of age. Control animals (N = 10) showed no consistent alteration in pituitary responsiveness to LHRH during development. Ewes treated with melatonin (N = 10) had puberty onset delayed by 4 weeks (P less than 0.03) but no effect of melatonin on LH or FSH response to LHRH injection was observed at any stage of development. In the control and melatonin-treated ewe lambs the responses to LHRH injection were lower during darkness than during the day at all stages of development. No consistent differences in LH or FSH pulsatility were observed between treatment groups or during development. Prolactin concentrations, however, failed to decrease at the time of puberty (autumn) in the melatonin-treated group. Melatonin-treated ewe lambs maintained normal rhythmic melatonin production which was superimposed on a higher basal concentration and showed the same increase in melatonin output with age as the control ewes. These results indicate that the delayed puberty caused by melatonin implants is not due to decreased pituitary responsiveness to LHRH or to dramatic changes in basal LH or FSH secretion.     

Effect of melatonin on pulsatile release of luteinizing hormone in female lambs.

This study investigated the effect of melatonin treatment of ewe lambs on LH pulsatility in an attempt to examine the mechanism whereby melatonin advances the onset of puberty. Six ewe lambs were given intravaginal melatonin implants at 12.8 weeks of age. Another six lambs received empty implants. All lambs were serially blood sampled every 15 minutes for six hours on several occasions prior to the onset of puberty. One week after implantation LH pulse frequency and mean LH levels were higher in treated lambs than the control lambs (pulse frequency 0.13/h vs 0.03/h; mean LH levels 2.0 +/- 0.2 ng/ml vs 1.3 +/- 0.1 ng/ml; p less than 0.05). Melatonin treatment failed to alter pulse frequency after the initial increase. Puberty was advanced by 3 weeks in the treated group. In the second experiment six lambs received melatonin implants at 13 weeks of age and another six lambs served as control. In this experiment blood samples were taken intensively during the first few weeks after treatment. Results of this study show that mean plasma LH levels and LH pulse frequency were again higher during the first week after implantation. This transient increase in LH release may be part of the mechanism initiating the eventual advancement of puberty although the significance of this increase is questionable. In both experiments the LH response to estradiol injection was monitored at various times after treatment, but no effects of melatonin were found, although the magnitude of the response increased with age.     

Ovarian response to hCG injections during the prepubertal period in three breeds of sheep with different ovulation rates and litter sizes

A positive correlation was reported between ovulation (OR) rate in response to hCG in prepubertal ewe lambs and adult ewes. The finding suggested that the response to hCG may be used to select prepubertal ewe lambs for prolificacy. We studied plasma FSH concentrations and ovarian response to hCG injection during the prepubertal period in breeds with high (Chios), medium (Florina) and low (Karagouniki) OR and litter sizes (LS). The results showed that Chios ewe lambs presented a higher profile of plasma FSH during the 3rd to 7th wk of age when compared to the breeds with medium (Florina) and low (Karagouniki) OR and LS. The ovarian response to hCG showed that the Chios ewe lambs responded earlier, by exhibiting ovulation at 6 wks and more effectively, by demonstrating a higher OR than the other two breeds. There was no correlation among plasma FSH concentrations during prepubertal age (3rd to 7th wk), ovulation rate after hCG injection, or litter size at first and second lambing for all breeds. Therefore we concluded that the plasma FSH concentrations and/or the ovarian response to hCG injection during the prepubertal period is not a suitable criterion for early selection for litter size in the adult ewe of these breeds.     

Inhibin B,follicle stimulating hormone,luteinizing hormone,and estradiol and their relationship to the regulation of follicle development in girls during childhood and puberty

Inhibin B, produced by granulosa cells in the ovary, is a heterodimeric glycoprotein suppressing synthesis and secretion of the follicle stimulating hormone (FSH). The aim of the present study was to determine hormone profiles of inhibin B, FSH, luteinizing hormone (LH), and estradiol in girls during childhood and puberty and to evaluate whether inhibin B is a marker of follicle development. We examined the correlation between inhibin B and gonadotropins and estradiol during the first two years and across the pubertal development. Using a specific two-side enzyme-linked immunosorbent assay (ELISA), inhibin B levels were measured in the serum of 53 healthy girls divided into 8 groups according to age. In addition, serum FSH, LH, and estradiol were measured by chemiluminescent immunoassay in all serum samples. A rise in serum levels of inhibin B (55.2+/-7.3 ng/l, mean +/- S.E.M.) and FSH (1.78+/-0.26 UI/l), concomitant with a moderate increment of serum LH (0.36+/-0.09 UI/l) and estradiol (45.8+/-12.2 pmol/l) concentrations was observed during the first three months of life and declined to prepubertal concentrations thereafter. A strong positive correlation between inhibin B and FSH (r = 0.48, p<0.05), LH (r = 0.68, p<0.001) and estradiol (r = 0.59, p<0.01) was demonstrated during the first 2 years of life. A rise in serum levels of inhibin B, FSH, LH, and estradiol was found throughout puberty. Inhibin B had a strong positive correlation with FSH (stage I of puberty: r = 0.64, p<0.05; stage II of puberty: r = 0.86, p<0.01), LH (I: r = 0.61, p<0.05; II: r = 0.67, p<0.05), and estradiol (II: r = 0.62, p<0.05) in early puberty. From pubertal stage II, inhibin B lost this relationship to gonadotropins and estradiol. Serum inhibin B and FSH levels increased significantly during pubertal development, with the highest peak found in stage III of puberty (133.5+/-14.3 ng/l), and decreased thereafter. In conclusion, inhibin B is produced in a specific pattern in response to gonadotropin stimulation and plays an important role in the regulation of the hypothalamic-pituitary-gonadal axis during childhood and puberty in girls. Inhibin B is involved in regulatory functions in developing follicles and seems to be a sensitive marker of ovarian follicle development.     

Changes in episodic luteinizing hormone secretion leading to puberty in the lamb

In this study, we monitored episodic luteinizing hormone (LH) secretion throughout development in eight April-born ewe lambs to determine if a change in LH pulse patterns preceded first ovulation at puberty. LH pulses were measured in samples collected every 12 min for 6 h once in July, twice a month from 22 August to 2 October, and then weekly until puberty. Progesterone concentrations, measured in samples taken 3/wk, were used as an index of first ovulation, which occurred at 29.3 +/- 0.7 wk of age. LH pulse frequencies throughout most of this period ranged from 0 to 2 pulses/6 h, with no change over time. However, during the week prior to the first progesterone rise, there was a significant increase in pulse frequency to a level seen during the follicular phase in post-pubertal lambs. This increase in pulse frequency was evident in 7 of 8 lambs; pulses were not analyzed in the last lamb because samples were taken during the LH surge. In contrast, LH pulse amplitude did not increase prior to puberty. In fact, pulse amplitude declined linearly during the 3 wk before first ovulation and then increased during the follicular phase in post-pubertal animals. These results support the hypothesis that an increase in the frequency of episodic LH secretion is a key event leading to the onset of ovarian cycles in the lamb. Whether an increase in pulse amplitude is also necessary remains unclear. If so, it must occur just before the LH surge, since it was not detected in any samples taken before puberty in this study.     

Induction of precocious puberty in ewe lambs by pulsatile administration of GnRH

Circhoral administration (250 ng/h, i.v.) of GnRH induced a preovulatory-like surge of LH and subsequent luteal function in 4 of 4 ewe lambs 1 month before expected date of puberty. Within 12h of the start of pulsatile delivery of GnRH, mean concentrations of immunoactive and bioactive LH increased significantly (P less than 0.05) and the LH surge occurred by 1.8 +/- 0.6 days of treatment. Mean concentrations of serum progesterone were elevated significantly (P less than 0.001) 3 days after the surge. The biopotency of LH (bioactive LH/immunoactive LH) before the GnRH-induced surge of LH did not differ from LH biopotency in ewe lambs receiving circhoral delivery of saline (0.41 +/- 0.05 and 0.46 +/- 0.04, respectively). Biopotency of LH declined markedly at the GnRH-induced LH surge (0.25 +/- 0.04), but biopotency of serum LH was significantly augmented (P less than 0.05) during the period of luteal activity (0.70 +/- 0.07). Regular oestrous cycles were observed in 3 of 4 ewe lambs after the 10-day GnRH treatment period. These results indicate that pulsatile delivery of GnRH is effective in inducing precocious puberty in ewe lambs. Increase in LH biopotency does not appear to be required in the pubertal transition to reproductive cyclicity in this species. Augmented LH biopotency may be important in support of luteal function after first ovulation.     

Patterns of circulating gonadotropins and ovarian steroids during the first periovulatory period in the developing sheep.

This report provides evidence that an increment in serum gonadotropin levels occurs at puberty in the sheep and that this reflects the critical hormonal event culminating in first ovulation in this species. Blood samples were collected from 6 female lambs at 4-h intervals for a period of approximately 2 mo around the expected time of puberty (32 wk of age) until behavioral estrus was observed and ovulation was verified by assay of serum progesterone. Patterns of circulating LH, FSH, progesterone, and estradiol concentrations were characterized during the peripubertal period for each lamb. A rise in serum levels of both LH and FSH began approximately 7-10 days before the first preovulatory surge of gonadotropins. Although the increase in gonadotropin levels occurred gradually over several days, serum estradiol levels rose only during the final 40-60 h prior to the preovulatory surge of gonadotropin. Serum progesterone profiles revealed, however, that normal (14-16-day) luteal phases were induced in only 2 of 6 females as a result of the first surge. In four lambs, a short luteal phase of 2.5 days' duration occurred, which was followed by another estradiol rise and a preovulatory surge that then resulted in a full luteal phase of 14 days' duration. These data demonstrate clearly that the precipitating event at puberty in the female sheep is an increase in circulating gonadotropin levels and that the estradiol secreted from the newly stimulated follicle provides the signal for the first preovulatory surge.     

Bioactive and immunoreactive FSH concentrations in ewe and ram lambs over the first year of life

Several studies suggest that the concentration of immunoreactive (I) FSH measured in peripheral plasma by radioimmunoassay does not always reflect the level of bioactive (B) hormone capable of eliciting a biological response (e.g. oestradiol synthesis by Sertoli cells in vitro). The aim of this study was to measure both B-FSH and I-FSH concentrations in male and female sheep during the first year of life, and to relate this to pubertal development. The hypothesis being tested was that B-FSH is present in both male and female sheep during the prepubertal period and that discrete changes in B-FSH are associated with the onset of puberty. Eight ewe lambs and eight ram lambs were blood sampled fortnightly from 2 to 52 weeks of age. All samples were assayed for B-FSH and I-FSH content. Pubertal development was monitored in ewe lambs from behavioural oestrus and from plasma progesterone concentrations, and in ram lambs from penile and testicular development and from plasma testosterone concentrations. Mean I-FSH concentrations varied significantly with time after birth, in both females and males (P<0.01). In contrast, B-FSH was found to vary with time in females only (P<0.01). Around the expected time of puberty in ram lambs (i.e. at 30–40 weeks of age), and thereafter, I-FSH concentrations were undetectable (<0.2 ng ml⁻¹), whereas the B-FSH concentrations were measurable at concentrations up to twice the assay detection limit (0.8 ng ml⁻¹) until 38 weeks of age. In ewe lambs, but not ram lambs, there was a significant linear relationship between B-FSH and I-FSH values (R=0.595; P<0.005). When standardised about the time of puberty, B-FSH (P<0.05) but not I-FSH was significantly higher in ewe lambs that failed to reach puberty. No differences for either B-FSH or I-FSH between pubertal and non-pubertal ram lambs were noted. In summary, B-FSH was often measurable in plasma throughout prepubertal development in sheep and the concentrations often differed from those of I-FSH, especially in ram lambs. However, there appeared to be no discrete changes in B-FSH that could be directly related to specific pubertal events. It is concluded that although FSH may be a prerequisite for prepubertal testicular development and/or ovarian follicular growth, it is not a critical factor in determining whether puberty is attained during the first year of life in this seasonally breeding species.     

Plasma FSH and LH in prepubertal Booroola ewe lambs

Basal plasma concentrations (four 30-min samples) and GnRH-induced release of gonadotrophins were measured every 15 days between 30 and 90 days and at 110 days of age in Merino ewe lambs from the prolific Booroola ('B') flock (n = 18-23), the medium prolificacy ('T') flock (n = 14-20), and the 'O' flock (n = 4-8) of low prolificacy. At ages of 30 and 45 days B ewe lambs had mean basal plasma FSH concentrations of 145 and 122 ng/ml which were significantly higher (P less than 0.01) than those seen in T (45 and 53 ng/ml), and O (39 and 38 ng/ml) flock ewes. Between 60 and 110 days of age there were no significant differences between genotypes. The increment in FSH concentrations above basal levels induced by the subcutaneous injection of 100 micrograms synthetic GnRH was only significantly (P less than 0.05) greater in B than T and O genotype ewe lambs at 110 days of age but not at other ages. The basal plasma FSH differences between the B, T and O genotypes at 30 and 45 days of age were not consistently related to the size of litter in which lambs were born. At 30 days of age the mean plasma LH concentration of B, T, and O flock lambs were 2.6 +/- 0.5, 1.2 +/- 0.6 and 0.7 +/- 0.8 ng/ml respectively. These differences were not significant. At later ages there were also no significant differences between the genotypes with respect to basal LH, and the increase in LH induced by exogenous GnRH was always similar for the three genotypes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Antral follicle growth and endocrine changes in prepubertal cattle,sheep and goats

In the growing heifer calve, there is an early post-natal, gonadotrophin driven increase in ovarian antral follicle growth. The endocrine regulation of and reason for this initial stimulation of ovarian follicular development are not fully understood. This initial endocrine activity appears to be later held in check by negative feedback suppression mechanisms until the heifer is of a sufficient body size to initiate oestrous cycles and to reproduce. There is increasing evidence from recent ultrasonographic studies, performed in the same groups of prepubertal heifer calves, that the development of ovarian antral follicles and tubular genitalia occur in parallel. There appear to be two distinct periods of enhanced development of the reproductive organs, from 2 to 14 weeks of age and again from 34 to 60 weeks of age, or just prior to puberty. First ovulation in heifers is preceded by a gradual increase in pulsed LH secretion, which results in enhanced antral follicle development and oestrogen production. It was demonstrated that prepubertal heifers produced recurrent antral follicular waves; maximum sizes and life span of the dominant follicles of waves, as well as periodicity and FSH dependency of wave emergence were similar to those in adult cattle. In does, no Graafian follicles are seen at birth and total follicle numbers increase to 2 months of age, and then decline to 5 months of age. In ewe lambs, studies using transrectal ovarian ultrasonography showed that antral follicle recruitment and growth increased after the first 2 months of age and just before puberty. This bi-phasic pattern of changes in ovarian follicle recruitment and growth is strikingly similar to that in heifer calves, but it contrasts with earlier post-mortem examinations of ovaries in ewe lambs. Unlike in cattle and adult ewes, the rhythmic pattern of follicular wave emergence was not established in pre- and peripubertal ewe lambs. The early increase in antral follicle numbers and size in ewe lambs may be, at least in part, due to changes in FSH release and potency, and enhanced follicle production prior to first ovulation is probably caused by an increase in the frequency of LH pulses.     

Pituitary response to GnRH and ovariectomy in Booroola-Awassi and Awassi ewe lambs

Booroola-Awassi ewe lambs were heterozygous (F+) for a major gene F, influencing their ovulation rate, while Awassi lambs were non-carriers (++). Basal plasma FSH concentration (mean +/- s.e.m.) in Booroola-Awassi ewe lambs at 4 weeks of age was significantly higher than in Awassi lambs of the same age (5.06 +/- 0.60 and 2.04 +/- 0.32 ng/ml respectively; P less than 0.001). After GnRH administration, FSH increased from 3.89 +/- 1.10 to 10.58 +/- 1.30 ng/ml in Booroola-Awassi (N = 6) and from 1.87 +/- 0.29 to 4.64 +/- 0.33 ng/ml in Awassi (N = 6) ewe lambs (P less than 0.05). Ovariectomy caused an increase in plasma FSH in Booroola-Awassi (N = 4) and Awassi (N = 4) ewe lambs. At 1 week after ovariectomy plasma FSH increased from 5.96 +/- 1.02 to 7.06 +/- 1.05 ng/ml in F+ and from 1.67 +/- 1.06 to 5.21 +/- 0.66 ng/ml in ++ ewe lambs, suggesting a stronger negative feed-back effect exerted by the ovaries of Awassi lambs. At 15 weeks after ovariectomy FSH values were similar in Booroola-Awassi (18.28 +/- 1.96 ng/ml) and Awassi (16.07 +/- 0.70 ng/ml) lambs. Although the overall pattern of pituitary response to ovariectomy was similar in the F+ and ++ ewe lambs, Booroola-Awassi lambs had small ovaries (132.5 +/- 24.9 mg) and follicular development did not proceed beyond the preantral stage in 3/4 animals, and Awassi lambs had large ovaries (600.0 +/- 233.9 mg) (P less than 0.05) with many preantral and antral follicles.(ABSTRACT TRUNCATED AT 250 WORDS)     

Duration of oestrus, ovulation rate, time of ovulation and plasma LH, total oestrogen and progesterone in Galway adult ewes and ewe lambs

The mean duration of oestrus, ovulation rate, duration of the preovulatory LH discharge, time interval between sponge removal and beginning of the LH discharge, total LH discharged, maximum LH value observed and the concentration of progesterone in the peripheral plasma during the luteal phase of the oestrous cycle was similar in Galway adult ewes and 8-month-old ewe lambs after treatment with intravaginal sponges containing 30 mg cronolone for 12 days and injection of 500 i.u. PMSG. The interval between sponge removal and the onset of oestrus was shorter for adults than for ewe lambs; the interval between the onset of oestrus and the beginning of the LH discharge was longer in adults. During the period 12-36 h after sponge removal the mean plasma total oestrogen concentration was significantly higher in lambs than in adults. In a separate study of the time of ovulation in Galway ewe lambs given the same progestagen-PMSG treatment, ovulation did not occur in any lamb before 17 h after the onset of oestrus and the majority ovulated close to the end of oestrus.     

Sexual maturation of ewes raised without ram exposure in a controlled lighting environment

Finn x Dorset ewe lambs (n = 70) born in the spring (March 28 to April 6) from two successive lambing seasons were evaluated for age at first ovulation in the absence of mature rams. Ewe lambs were born in a controlled, short light (8L:16D) photoperiod or in ambient light (13L:11D). At about 10 to 11 wk of age, ewe lambs were allocated to a short (8L:16D) or long (16L:8D) light environment. Plasma progesterone (P(4)) concentrations were measured as an index of first ovulation. First exposure of ewes to sexually mature rams was in November. Most ewe lambs (77%) ovulated before ram exposure. More lambs (P < 0.025) born in ambient light and raised in short light reached puberty with typical cycles of plasma progesterone compared to other treatments. Long days tended to retard the onset of puberty. Although pregnancy rate did not differ across light treatments, more ewes became pregnant from the ambient-light born and short-light raised treatment. Photoperiod is an important factor affecting the onset of sexual maturation and genesis of normal luteal progesterone secretion in the ewe lamb.