Induction of ovulation in chronically hypophysectomized Booroola ewes

Booroola Merino ewes, with (F+; N = 17) and without (++; N = 13) a copy of the fecundity gene were hypophysectomized and 6 weeks later were given an i.m. injection of PMSG (high, medium or low dose) followed by hCG. The induced ovulation rates were observed laparoscopically. Ovulation rates were significantly higher (P less than 0.01) in Booroola F+ ewes than in ++ ewes (8.00 +/- 1.66 s.e.m. vs 3.62 +/- 1.10 respectively). This suggests that the high fecundity of the Booroola ewe may be due primarily to ovarian rather than pituitary factors.     

Natural and induced ovulation rate in prolific and non-prolific breeds of sheep in Ireland, Morocco and New Zealand

Ovulation rate, in mixed-age groups of prolific and non-prolific ewe breed types, after administration of a range of doses of PMSG (0, 375, 750 and 1500 i.u.) during the follicular phase of the oestrous cycle, were compared in Ireland, Morocco and New Zealand. The ewes in Ireland and Morocco were from the Finnish Landrace and Galway, and D'Man and Timhadite breeds, respectively. In New Zealand Booroola Merino x Romney ewes which had been previously identified as heterozygous carriers (F+) of the Booroola high fecundity gene and purebred Romneys were used to represent the prolific and non-prolific genotypes respectively; in addition a group of Booroola Merino x Romney non-carriers (++) of the major gene were also included for comparison. Ovulation rate at the oestrus which preceded stimulation with PMSG was also measured in all animals. In all 3 locations the ewes of the prolific genotype had a greater ovulation rate after PMSG stimulation than did the non-prolific controls. However, this association between prolificacy and response to PMSG was removed when ovulation rate after PMSG was transformed by dividing by the ovulation rate observed before PMSG administration. Despite the differences in the genetic basis of their high prolificacy the pattern of response to PMSG over the range of dosages used was similar in Finnish Landrace, D'Man and Booroola Merino x Romney (F+) ewes and all breeds had means of about 10 ovulations in response to 1500 i.u. PMSG. Amongst the non-prolific breeds, the Timhadite was the most responsive to PMSG although it had the lowest natural ovulation rate.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of long-term hypophysectomy on ovarian follicle populations and gonadotrophin-induced adenosine cyclic 3',5'-monophosphate output by follicles from Booroola ewes with or without the F gene

Long-term (i.e. approximately 70 days) hypophysectomy led to a significant (P less than 0.05) reduction in ovarian weight but no reduction in the total number of antral follicles (greater than 0.1 mm in diameter). In hypophysectomized ++ Booroola ewes (N = 8) follicles were always less than or equal to 3 mm and in hypophysectomized FF Booroola ewes (N = 6) follicles were always less than or equal to 2 mm in diameter; in ewes of both genotypes follicles reached diameters which were approximately 40% of their predicted final size at ovulation. Under in-vitro conditions, follicles from the FF and ++ hypophysectomized ewes produced significant increases in cAMP within 1 h of exposure to gonadotrophins (P less than 0.05) although no genotypic differences in cAMP production were noted. We conclude that ovarian follicles in FF and ++ ewes have absolute requirements for pituitary hormone on reaching diameters of 2 mm and 3 mm respectively and that appreciable numbers of antral follicles in ewes of both genotypes remain responsive to pituitary gonadotrophins despite prolonged deprivation of these hormones.     

Variations in patterns of follicle development in prolific breeds of sheep

Prolific breeds of sheep (Romanov, Finn and Booroola Romanov crosses heterozygous for the Booroola gene (F+) were compared with breeds of lower prolificacy (Ile-de-France, Finn X Scottish Blackface, Merino X Blackface and Booroola X Romanov not carrying a copy of Booroola gene (++] by in-vivo monitoring of follicular kinetics by ink labelling during the late luteal phase and follicular phase of the oestrous cycle followed by histological examination of the ovaries or follicle dissection. At each of 3 successive laparotomies, the 3 largest follicles of each ovary were measured and ink labelled. At the final laparotomy, around the beginning of oestrus, all ewes were ovariectomized. High ovulation rate was not associated with the total number of antral follicles in any of the breeds. However, there were more follicles greater than 2 mm in diameter in Romanov and Booroola X Romanov crosses (F+) compared to their respective controls. Such a feature was not observed in Finnish Landrace compared to Finn X Blackface and Merino X Blackface ewes. A more numerous population of recruitable follicles, together with a similar incidence of selection through atresia, were the features associated with the high ovulation rate of Romanov compared to Ile-de-France ewes. The high ovulatory potential of the Finn ewes resulted from a markedly reduced incidence of selection through atresia. Booroola X Romanov ewes carrying a copy of the Booroola gene (F+) appeared to possess features of both parental breeds, including high numbers of recruitable follicles, smaller follicular size when recruitment occurs and an extended time for recruitment. Booroola X Romanov (++) ewes, not carrying the gene, appeared to have lost part of the 'Romanov characteristics' of a more numerous population of recruitable follicles. The variability in the kinetics of preovulatory enlargement, seen in these breeds of sheep, demonstrates that there are a number of pathways through which high ovulation rate can be achieved and hence through which ovulation rate might be manipulated.     

Fertilization and embryo loss in Booroola Merino x South Australian Merino ewes: Effect of the F gene

The effects of Booroola genotype (F+, ++); the number of ovulations per ewe (one, two or three); and the age of a ewe (2.5 yr vs 3.5 to 6.5 yr) on the percentage of ova fertilized, embryo loss and fetal loss were examined in Booroola x South Australian Merino ewes slaughtered on Days 4, 21 and 90 after insemination. Ewes slaughtered on Day 90 were examined by real-time ultrasound imaging (RUI) on Day 45. Fertilization failure was independent of ewe genotype, ovulation rate and age of ewe, and it was not an important source of wastage (F+, 9.4%; ++, 6.7%). Most embryo loss occurred during the first 21 d (F+, 54.7%; ++, 40.3%). Interpretation of the effects of genotype and ovulation rate on embryo wastage measured on Days 21, 45 and 90 was obscured by significant (P < 0.05) genotype and ovulation rate interactions with the day of slaughter/RUI. The effect of age on embryo loss was not significant (P > 0.05). Reasons for the high rate of wastage observed in this experiment require further study.     

Effect of exogenous melatonin and plane of nutrition after weaning on estrous activity, endocrine status and ovulation rate in Salz ewes lambing in the seasonal anestrus

Forty-nine Spanish Salz ewes lambing in the second fortnight of March (20 March +/- 1.5 d) were used to determine the effects of exogenous melatonin and postweaning nutrition on endocrine status, date of first estrus and ovulation rate. Experimental design was a factorial defined by 2 postweaning planes of nutrition, 1.80 (high) and 1.35 (low) times the maintenance requirements, and treatment with a single 18-mg subcutaneous implant of melatonin (M) 32 d after lambing or no treatment control (C). Mean weaning to first estrus interval was shorter in treated than in control ewes (50.8 +/- 4.2 vs 87.6 +/- 6.3 d; P < 0.01). Considering both the treated and control animals together, the ratio between mean night and daytime plasma melatonin levels was significantly correlated with the implant insertion-first estrus interval on Day 5 (0.67; P < 0.01) and Day 35 (0.63; P < 0.05) after implantation. Melatonin implants induced a significant increase of mean LH concentrations at Days 14 and 33 after implantation (P < 0.01) without any significant influence of plane of nutrition. Ovulation rate was higher for treated than control ewes in the second estrus (P < 0.05). An interaction between plane of nutrition and exogenous melatonin on ovulation rate at the second cycle after weaning was detected (P < 0.01), being close to the significance in the first, fourth and fifth cycles (P < 0.1). These results suggest that exogenous melatonin in April may be an effective way of advancing the breeding season and enhancing ovulation rate associated with a low rather than a high plane of nutrition.     

Binding characteristics of 125I-labelled human FSH to granulosa cells from Booroola ewes which were homozygous, heterozygous or non-carriers of a major gene(s) influencing their ovulation rate

At 37 degrees C 125I-labelled human (h) FSH (NIAMDD-hFSH-I-3) bound rapidly to granulosa cells from Booroola and Romney ewes with 50% maximum binding achieved after 3 min and equilibrium being reached within 45 min, irrespective of whether the cells were obtained from the FF, F+ or ++ Booroola genotypes or from Romney ewes. Binding of 125I-labelled FSH followed second order kinetics and there was no effect of follicle diameter (1-2.5 mm vs greater than or equal to 3 mm). Irrespective of breed, genotype or follicle size, the mean (+/- s.e.m.) calculated association rate constant, (ka) was 7.3 (+/- 0.8) x 10(5) litres mol-1 sec-1 (n = 12). Dissociation of receptor bound 125I-labelled hFSH was less than 5% after 30 min and low but variable (i.e. between 0 and 30%) after 2-6 h irrespective of breed, genotype or follicle size. No gene-specific differences were noted in binding specificity between F+ and ++ genotypes: studies were not performed with cells from FF ewes because of insufficient cells. The binding of 125I-labelled hFSH could be displaced with sheep FSH (NIH-FSH-S16; 10% cross-reaction) and FSH-P (2.5% cross-reaction) but other sheep pituitary hormones and hCG showed little or no cross-reaction (less than or equal to 0.1%). The calculated binding capacities (Bmax) and equilibrium dissociation constants (Kd) for 125I-labelled hFSH binding to granulosa cells did not differ between the Booroola genotypes or between Booroola or Romney follicles of different diameter (i.e. 1-2.5 mm; or greater than or equal to 3 mm). The overall mean +/- s.e.m. (n = 24) Bmax and Kd values were 16.7 +/- 0.8 fm/mg protein (i.e. approximately 800 available receptor binding sites/cell) and 1.1 +/- 0.1 nM respectively. Collectively, these findings suggest that the earlier maturation of follicles in FF or F+ ewes compared to ++ ewes is unlikely to be due to gene-specific differences in the FSH binding characteristics of the granulosa cells.     

GnRH-induced gonadotrophin secretion in ovariectomized Booroola ewes with hypothalamic-pituitary disconnection

To test whether the F gene-specific differences in the plasma concentrations of FSH and LH are due to differences in the pituitary responsiveness to exogenous GnRH, ovariectomized Booroola ewes with hypothalamic-pituitary disconnection (HPD-ovx) were treated with GnRH (250 ng i.v.) once every 2 h for up to 5 weeks. In Exp. 1, jugular venous blood was collected once weekly from 13 FF and 14 ++ HPD-ovx ewes for 6 weeks before GnRH treatment and every 2nd, 3rd or 6th day for 5 weeks during treatment. In Exp. 2, jugular venous blood was collected from another 8 FF and 7 ++ HPD-ovx ewes at 5- or 10-min intervals over 4 GnRH pulses (250 ng i.v. once every 2 h) on 3 separate occasions after the animals had been subjected to the GnRH pulse regimen for approximately 7 days beforehand. Also in Exp. 2, the animals were extensively sampled around a larger (10 micrograms) i.v. injection of GnRH and the pituitary FSH and LH contents assessed after the animals had been re-exposed to the once every 2 h GnRH (250 ng i.v.) pulse regimen for several days following the larger GnRH bolus. In Exp. 3 the distributions of mean plasma concentrations of FSH and LH in individual GnRH-treated HPD-ovx ewes were compared with those in ovariectomized and ovary-intact FF and ++ ewes. During the 6 weeks before GnRH treatment (Exp. 1), the plasma concentrations of FSH (approximately 1 ng/ml) and LH (less than or equal to 0.8 ng/ml) were not different between the genotypes. After GnRH treatment both the mean FSH and LH concentrations increased significantly (P less than 0.01) above basal values after 2 days with F gene-specific differences being noted for FSH but not LH (FSH; FF greater than ++; P less than 0.05). Thereafter, the mean FSH but not LH concentrations increased at a faster rate in FF than in ++ ewes with the overall mean FSH concentrations between the genotypes being significantly different (P less than 0.05). In Exp. 2 considerable between-animal variation in the pulsatile pattern of FSH but not LH concentrations was seen in ewes of both genotypes during GnRH treatment. The overall mean FSH concentrations were higher in FF than in ++ ewes (P less than 0.05) and the mean FSH response to each GnRH pulse was significantly higher in FF than in ++ ewes (P less than 0.05).(ABSTRACT TRUNCATED AT 400 WORDS)     

Pre-ovulatory follicular characteristics and ovulation rates in different breed crosses, carriers or non-carriers of the Booroola or Cambridge fecundity gene

Terminal follicular dynamics and ovulation rates (OR) were compared in different local breeds after introducing fecundity genes of different origin. Crossbred ewes which were carriers (F+) or non-carriers (++) of Booroola (BFec) or Cambridge genes (CFec) were included: CambridgexCambridge (CC), CambridgexSuffolk (CS), CambridgexTexel (CT), BooroolaxTexel (BT) and BooroolaxGerman Mutton Merino (BGM). The numbers of small (diameter 2-3.5 mm), medium (diameter >3.5-5.0 mm) and large (diameter >5.0 mm) growing follicles, the maximum diameter before ovulation and the regression and artesia rates of ovarian follicles >/=2 mm in diameter were studied laparoscopically and repeatedly during the last 5 days of an induced oestrous cycle. The ORs were determined one cycle before and two cycles after the repeated laparoscopy. BFec and CFec significantly enhanced the OR of all crossbreeds. Carriers of BFec or CFec did not have significantly different ORs due to any crossbreeding effect. The same observation was made for non-carriers of both Fec gene types. Whatever the crossbreed, the number of small, medium and large growing follicles were similar between carriers and non-carriers in spite of a higher number of ovulating follicles in carriers of both Fec gene types. The diameter of ovulatory follicles did not differ among crossbreds, or between carriers and non-carriers except in the BT (5.2+/-0.2 vs. 6.5+/-0.8 mm, respectively) and CC (6.6+/-0.2 vs. 5.6+/-0.3 mm) ewes.The higher OR in the presence of the Booroola gene was associated with a low atresia rate of large follicles in all crossbreeds (BT: 52+/-8% (F+) vs. 61+/-7% (++); BGM: 51+/-6% vs. 75+/-5%). The high OR of the carriers of the CFec gene seemed to be associated with a lower number of large growing follicles with a lower (P<0.05) atresia rate as compared with Booroola crossbreeds.In conclusion, follicular features were similar between purebred Cambridge and its crossbred CS and CT. In ewes carrying the BFec or CFec gene, the reduction in follicular atresia seemed to be one of the main follicular features implicated in the higher OR.     

A review of the effects of the Booroola gene (FecB) on sheep production

Booroola Merino (B^oM) ewes have a high ovulation rate and litter size which in 1980 was postulated to be due to the effects of a major gene (FecB). This was confirmed in breeding experiments and FecB was subsequently shown to be due to a mutation (BMPR-1B) on chromosome 6. The B^oM originated from an Australian commercial fine wool Merino flock (Booroola) and has been used in crossing experiments and for introgression of FecB into many breeds around the world to improve fecundity. The mutation has recently been found in native sheep breeds in India, China and Indonesia and it is likely that FecB in the Australian B^oM was derived from importations of Garole sheep from India in 1792 and 1793.The effects on production traits of the FecB mutation in a range of genetic comparisons, environments and production systems are reviewed. Comparisons involving B^oM crosses with various other breeds and contrasts of FecB homozygous (BB), heterozygous (B+) and non-carrier (++) genotypes in comparable background genotypes, including non-B^oM, have been summarised from 45 reports. The weighted mean effect for ewes carrying one copy of FecB (B+) was +1.3 (range +0.8 to +2.0) for ovulation rate and +0.7 (range +0.4 to +1.3) for litter size. The effect of a second copy (BB) was generally additive for ovulation rate, with little or no increase in litter size for BB ewes among B^oM crosses. However there was generally a further increase in litter size for BB ewes of about half the effect of one copy (B+) in the Indian and Chinese breeds. Poor lamb survival and lamb growth reduced the number of lambs weaned and total weight of lamb weaned by B+ ewes. Most studies still showed a small advantage for B+ ewes, although several reported negative effects. While embryo survival declines at higher ovulation rates, the effects of FecB per se are equivocal. There is some evidence of a higher non-pregnancy rate among homozygous BB ewes. Most studies reported lower birth weight and growth rate from B^oM cross lambs and lambs from crossbred ewes introgressed with FecB. However it is difficult to separate the effects of low background genetic merit for growth of the B^oM and the lower birth weight and growth rate of lambs from larger litters from the genetic effect of carrying FecB. There was little or no difference in growth rate between BB, B+ and ++ genotype lambs. For other traits including, seasonal oestrous activity, carcass and meat quality and wool production, there was no evidence of major effects of FecB. The opportunities for management and nutritional modification of FecB expression and implications for industry adoption are briefly discussed.     

Immunoassay of gonadotrophin-releasing hormone in brain tissue of Booroola Merino ewes

The presence of a fecundity gene (F) in Booroola Merino ewes increases the ovulation rate. To test how F gene expression affects the gonadotrophin-releasing hormone (GnRH) concentration in hypothalamic or extrahypothalamic regions of the brain, GnRH was measured by radioimmunoassay in acetic acid extracts of various brain tissues from Booroola ewes which were homozygous (FF), heterozygous (F+) or non-carriers (++) of the F gene. The GnRH concentration in brain tissues from FF, F+ and ++ animals which had been ovariectomized 5 months previously was also evaluated. No significant F gene-specific differences were noted in any of the brain areas tested, in intact or ovariectomized animals. However, in ovariectomized ewes, the concentrations of GnRH increased about 2-fold in the median eminence of the hypothalamus, remained unchanged in the medial basal hypothalamus and dropped to less than 10% of the values in intact ++ animals in the preoptic area. These studies suggest that the changed pituitary sensitivity and increased gonadotrophin release in Booroolas carrying the F gene(s) is not attributable to increased hypothalamic GnRH concentrations in these animals.     

Reproductive biology of the Booroola Merino sheep

This paper reviews the genetic and physiological characteristics of the Booroola Merino, one of the four most prolific sheep breeds in the world, and which was acquired by CSIRO in 1958 from a commercial sheep property, 'Booroola', Cooma, N.S.W. The exceptional prolificacy of this genotype--e.g. mean flock ovulation rate in 1982 of 4.2 (range 1-10) and mean litter size of 2.5 (range 1-7)--is largely attributable to a single gene (F) of uncertain origin which increases ovulation rate. Crosses of the Booroola with other Merinos produce progeny which have a 47-87% increase in ovulation rate, a 45-56% increase in litter size at birth, and a 1-33% reduction in lamb survival relative to control Merinos. This represents a 16-37% increase in the number of lambs weaned per ewe joined in favour of the Booroola crosses. The exact site of action of the F gene is not well established, although it is expressed primarily at the ovary, where more than the normal number of follicles mature and ovulate each oestrous cycle. This may result from some abnormality of the Booroola follicle itself or it may reflect differences in Booroola gonadotrophin secretion. There is some evidence that Booroola ewes have elevated plasma concentrations of follicle stimulating hormone (FSH) early in life and during the oestrous cycle, and that FSH concentrations in the pituitary gland and urine of the adult ewe are also high. These elevated FSH levels in the adult are attributed to an ovarian feedback deficiency, probably because the inhibin content of the Booroola ovary is only one-third that of normal Merino ovaries. The low inhibin content appears to be due to Booroola follicles having significantly fewer granulosa cells than control Merinos. Analogous studies of the prolific D'man sheep of Morocco point to FSH as the main correlate of prolificacy. The testis growth rate, testis size and total daily production of spermatozoa of the Booroola ram are similar to those of normal Merinos, as also are the endocrine characteristics of adult rams. The Booroola gene's expression is evidently sex-limited. Several theories concerning the mode of action of the F gene are being tested.     

Ovarian follicular populations and preovulatory enlargement in Booroola and control Merino ewes

To investigate the factors contributing to the different ovulation rates observed in two strains of sheep (Booroola 5.2, Merino 1.2), in-vivo monitoring of follicular kinetics followed by histological examination of both ovaries was performed during the late luteal and follicular phases. Ewes of both strains were either ovariectomized at Day 13, or had the 3 largest follicles of each ovary ink-labelled at Day 13 and were ovariectomized at Day 15, or had the 3 largest follicles of each ovary ink-labelled at Days 13 and 15 and were ovariectomized 16 h after the beginning of oestrus (N = 6 per time per strain). In another experiment, the age effects on the follicular populations of these two strains were also studied. There were 2-4 times more primordial follicles and 1.5-2 times more preantral follicles in the ovaries of Booroola than in control Merino ewes, although the number of antral follicles was the same. The percentage of normal follicles in this population was higher in Merino than Booroola ovaries. In Booroola ewes, there was no correlation between the number of antral follicles per ovary and the ovulation rate at the previous cycle (r = 0.22). This suggests that follicle numbers do not play a key role in the high ovulation rate of the Booroola strain. The number of follicles initiating growth from the primordial pool, the number of growing follicles disappearing at the preantral stage, the pattern of antrum development, granulosa cell multiplication and appearance of atresia differed between strains. The reasons for the high ovulation rate of the Booroola strain became clear when preovulatory enlargement was followed by ink labelling. An extended period of time during which recruitment of ovulatory follicles takes place, together with a low incidence of selection and the ability of the follicles to wait for ovulation are the features involved in this high ovulation rate.     

The half-life of follicle-stimulating hormone in ovary-intact and ovariectomized booroola and control merino ewes

To determine whether the high ovulation rate of the Booroola Merino ewe could be explained by FSH metabolism we have tested the proposition that FSH may have a longer half-life in the plasma of Booroola Merino ewes than in control ewes. The half-life of plasma FSH was determined by removal of the pituitary gland, to abolish FSH secretion into the peripheral circulation, and monitoring by repeated blood sampling the subsequent decline in plasma FSH concentrations. The half-life of FSH was similar in Booroola (103 +/- 14 (s.e.m.) min, N = 8) and control (116 +/- 8 min, N = 9) ewes. However, when ewes that had been ovariectomized at least 6 months earlier were hypophysectomized, the half-life of FSH was increased from 110 + 8 min in ovary-intact ewes (N = 11) to 1101 +/- 49 min (N = 6) (P less than 0.001) with no difference between the two Merino strains. We conclude that changes in the circulating half-life of FSH do not account for the high fecundity of the Booroola but that ovariectomy can alter the half-life of FSH secreted by the pituitary gland.     

Concentrations of progesterone,follistatin, and follicle-stimulating hormone in peripheral plasma across the estrous cycle and pregnancy in merino ewes that are homozygous or noncarriers of the Booroola gene

The circulating concentrations of progesterone, FSH, and follistatin across the estrous cycle and gestation were compared in Australian merino sheep that were homozygous for the Booroola gene, FecB, or were noncarriers. The Booroola phenotype is due to a point mutation in the bone morphogenetic protein receptor 1B. Progesterone concentrations began to rise earlier and were higher in the Booroola ewes than in the noncarriers on most days of the luteal phase but not during the follicular phase of the cycle. Follistatin concentrations remained unchanged across the estrous cycle in both groups of ewes, with no differences between genotypes. FSH concentrations were higher in Booroola ewes than in noncarrier ewes on most days of the estrous cycle, with a significantly higher and broader peak of FSH around the time of estrus. Progesterone concentrations were significantly higher in early and midgestation in Booroola ewes but were lower toward the end of gestation than those in noncarriers. FSH declined in both groups across gestation, with lower concentrations of FSH in Booroola ewes during midgestation. Follistatin remained unchanged across gestation in Booroola ewes and noncarrier ewes with a twin pregnancy but declined across gestation in noncarrier ewes with a singleton pregnancy. These results suggest that follistatin concentration is not regulated by the FecB gene during the estrous cycle and pregnancy but is influenced by the number of fetuses. However, the FecB gene appears to positively affect both progesterone and FSH during the estrous cycle and across pregnancy, which suggests that bone morphogenetic proteins play an important role in the regulation of both hormones.     

Follicular dynamics and dominance in Booroola x Finnish Landrace and Booroola x Suffolk ewes heterozygous for the F gene

To study the influence of the F gene on follicular dynamics and dominance, 2-year-old Booroola x Finnish Landrace (BFL, N = 17) and Booroola x Suffolk (BS, N = 18) ewes were compared with contemporary purebred Finn (FL, N = 18) and Suffolk (S, N = 18) ewes. In Exp. 1, oestrous cycles of ewes were synchronized during the breeding season with progestagen-impregnated sponges. At sponge removal (Day 0), 14 days after insertion, ewes of each of the 4 genetic groups were assigned to Group 1 in which all follicles visible on both ovaries were destroyed by electrocauterization except for the largest (F1) which was marked, Group 2 in which all visible follicles on both ovaries were destroyed, or Group 3 in which the 3 largest follicles of both ovaries were identified as F1, F2 and F3 and marked. At 48 h after treatment (Day 2), follicular growth was evaluated. At Day 0, the mean number of small follicles (1-3 mm) was higher (P less than 0.05) for BS, S and BFL (35.8, 35.1 and 32.9) than FL (24.9) ewes. Large follicles (greater than or equal to 4 mm) were more numerous (P less than 0.05) in FL (3.5) than in BS (2.1) ewes, BFL and S ewes being intermediate. Diameter of the F1 follicle was larger (P less than 0.05) for S (7.6 mm) than FL, BS and BFL (5.8, 5.1 and 5.1 mm) ewes. In Group 1, all F1 follicles marked at Day 0 ovulated at oestrus after sponge removal for BFL, BS and S ewes while in FL ewes, 2 of 6 F1 follicles regressed. In ewes ovulating, only the F1 follicle ovulated except for one S ewe which shed one more ovum. In Group 2, there were no follicles greater than or equal to 4 mm at Day 2 and no ewes ovulated after treatment. In Group 3, the proportion of marked follicles that ovulated was higher for S ewes than in those of the prolific genotypes. The number of follicles not marked at Day 0 but ovulating (compared to the total number of ovulations) was higher in BFL, BS and FL (8/11, 9/13 and 9/13) than S (3/10) ewes. In Exp. 2, prolific (BFL + BS) and non-prolific (S) ewes were compared following destruction of follicles greater than or equal to 3 mm with the F1 left intact (Treatment 1) or destroyed (Treatment 2), 12 days after sponge insertion.(ABSTRACT TRUNCATED AT 400 WORDS)     

Effects of nutritional supplements on testicular size and the secretion of LH and testosterone in Merino and Booroola rams

Six Booroola and six Merino rams were fed either a diet which maintained constant live weight or the same diet plus a supplement of high protein lupin grain for 15 weeks, and changes in live weight and testicular volume were measured. Serial blood samples taken for 24 h before the start and 9 weeks after the treatment began were assayed for plasma LH and testosterone and the resulting profiles were analysed for pulses of both hormones. Five weeks later, the animals were given two intravenous injections of 1 μg gonadotrophin-releasing hormone (GnRH) 1 h apart in order to measure pituitary gland responsiveness. A further week later the animals were injected intravenously with 500 μg human chorionic gonadotrophin (hCG) and the levels of testosterone were measured in samples taken after 1.5 h to estimate the testicular responsiveness.The nutritional supplement stimulated testicular growth in both genotypes, so that at the end of the treatment period the testes had increased significantly (P<0.01) in volume by 66% in the Merinos and by 63% in the Booroolas. The live weights also increased, but by relatively less (34% and 43% for supplemented Merinos and Booroolas). The rates of increase in both testicular size and live weight were similar for the two breeds. There were no significant effects of diet on the tonic secretion of LH or testosterone, or on responsiveness to GnRH or hCG.The intervals between LH pulses were significantly shorter (P<0.05) in Booroola rams than in Merino rams both before and after treatment (5.8 h vs. 11.6 h before treatment). The breed differences in LH secretion were mimicked by the testosterone profiles. In the Booroolas, five of the twelve LH profiles contained groups consisting of two to four individually identifiable pulses, each of which elicited a separate pulse of testosterone. A pulse group was observed in only one profile from the Merinos (P=0.06). There were no significant differences between the genotypes in any other parameter of LH or testosterone secretion, or in their responsiveness to GnRH or hCG.It was concluded that (i) nutritional supplements will stimulate testicular growth in both Merino rams and Booroola rams; (ii) the increase in testicular size does not appear to involve an increase in the responsiveness of the testis to LH; and (iii) there are both qualitative and quantitative differences between the genotypes in the patterns of secretion of LH and testosterone which may be associated with the differences in their fecundity.     

Ovarian follicular activity in Booroola lambs with and without a fecundity gene

The ovaries of 3-month-old Booroola lambs which were heterozygous carriers of a major gene (F) influencing the ovulation rate in mature ewes (i.e. F + lambs) were compared to those ofsimilarly-aged Booroola lambs which were non-carriers of the F-gene (i.e. ++ lambs). The ovaries of the F + Booroola lambs were significantly lighter (P less than 0.01) than those of ++ lambs even though the mean +/- s.e.m. number of follicles (greater than or equal to 1 mm diam.) in the F + lambs was greater than that in the ++ lambs (i.e. F + lambs, 30.2 +/- 2.5 follicles; ++ lambs, 18.4 +/- 1.2 follicles; P less than 0.01). In granulosa cells from non-atretic follicles (greater than or equal to 1 mm diam.) from F + and ++ Booroola lambs, FSH (NIAMDD-FSH-S16) doses of 100 and 1000 ng/ml caused significant stepwise increases (P less than 0.05) in cyclic adenosine 3',5'-monophosphate (cAMP) production compared to that achieved at FSH doses of 0 and 1 ng/ml or at any FSH dose in cells from atretic follicles. However, no significant differences in FSH-induced cAMP production were noted with regard to Booroola genotype or follicular diameter. None of the granulosa cell preparations from non-atretic follicles of 1-2.5 mm diameter from F + lambs (N = 13) or from non-atretic follicles of 1-4.5 mm diameter from ++ lambs (N = 16) responded to LH (NIAMDD-LH-S24; 10 or 1000 ng/ml) to produce significantly more cAMP than did the controls. In contrast, the granulosa cell preparations from non-atretic follicles of 3-4.5 mm diameter from F + lambs (N = 4) and from non-atretic follicles of greater than or equal to 5 mm diameter of ++ lambs (N = 4) produced significantly more cAMP (P less than 0.05) in response to LH (1000 and/or 10 ng/ml) relative to that in the controls. The theca interna from follicles of lambs of both genotypes had functional LH receptors as judged by the androstenedione responses to exogenous LH although no genotypic differences were noted. In F + lambs, the follicular fluid concentrations of testosterone but not oestradiol (i.e. in 1-4.5 mm diam. follicles) and granulosa cell aromatase activity (i.e. in 3-3.5 mm diam. follicles) were significantly higher (both P less than 0.05) than in corresponding follicles or cells from ++ lambs. Collectively the results suggest that the Booroola F-gene influences the composition and function of sheep ovaries before puberty.     

Seasonal reproduction in ewes selected on seasonal changes in wool growth

Romney and Perendale ewes were selected on the amplitude of seasonal wool growth. The ewes were fed a constant plane of nutrition and run with vasectomized rams. Ovarian activity was recorded by laparoscopy during 11 months. Ewes with a low amplitude of seasonal wool growth (Group L) had a 68% higher wool growth rate in winter and a 17% lower wool growth rate in summer compared with ewes with a high amplitude (Group H). There was no difference between the groups in the date of the first mating mark. Ewes in Group L entered anoestrus significantly later than did ewes in Group H; the difference was 11 days in the mean date of the last mating mark and 17 days in the mean date of the last ovulation. A significantly higher proportion of ewes in Group L ovulated during July to November. In addition, ewes in Group L had a significantly higher proportion of multiple ovulations throughout the experiment: on average the difference between the groups was 0.21. These results show that phenotypic selection for a low amplitude of seasonal wool growth resulted in a delay in the end of the breeding season associated with an increase in ovulation rate, suggesting independent effects on the beginning and end of the breeding season.     

Differences in ovarian activity between booroola X merino ewes which were homozygous, heterozygous and non-carriers of a major gene influencing their ovulation rate

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.