Booroola Fecundity Gene Research Articles

Effects of passive immunization against inhibin-peptide on secretion of follicle-stimulating hormone and ovulation rate in ewes carrying the Booroola fecundity gene.

Objectives of the study were to determine whether 1) inhibin negative feedback regulation of FSH secretion is diminished in ewes carrying a copy of the Booroola fecundity (FecB) gene and 2) differential FSH secretion is obligatory for expression of gene-specific differences in ovulation rate (OR). The approach was to compare FSH and ovulatory responses to passive immunoneutralization of inhibin in ewes with and without a copy of the FecB gene. Twenty-eight 2- to 3-yr-old ewes were assigned within genotype to antibody (alpha-IF-Ab) or control groups. Genotypes consisted of 3/4 Rambouillet x 1/4 Booroola ewes with one copy of the FecB gene (FecB+; 57 kg) and 3/4 Rambouillet x 1/4 Booroola ewes without the FecB gene (++; 59 kg). Estrus was synchronized during the breeding season using progesterone-releasing pessaries (CIDR-G). Pessaries were removed at 0 h. A single injection of alpha-IF-Ab or control solution was given at -48 h. Alpha-IF-Ab had been developed against a synthetic inhibin fragment matching the N-terminal region of ovine inhibin's alpha subunit. For injection, alpha-IF-Ab had been precipitated from ovine immune sera and concentrated. Blood samples were collected at 6-h intervals from -48 to 48 h, and laparoscopy was performed 14 days after CIDR-G withdrawal. All ewes exhibited estrus and ovulated. Genotype and alpha-IF-Ab treatment were without effect on intervals to estrus. Both factors affected OR (p < or = 0.001). Mean OR in control ++ and FecB+ ewes were 1.6 and 2.7, respectively; mean OR in alpha-IF-Ab-treated ++ and FecB+ ewes were 2.5 and 4.6, respectively. Following injection of alpha-IF-Ab, FSH concentrations increased within 6 h, peaked 12-18 h later, and then declined. Magnitude of FSH increases was similar in ++ and FecB+ ewes (70% and 85% over control values, respectively). Results demonstrate that 1) inhibin negative feedback regulation of FSH secretion is not a site of FecB gene action and 2) the mechanism by which the FecB gene increases OR does not necessarily involve increased FSH secretion during the period of preovulatory follicular development.

Expression of gonadotrophin subunit genes in sheep that were homozygous carriers and non-carriers of the Booroola fecundity gene FecB.

Homozygous carriers (BB) of the Booroola fecundity gene FecB are characterized by high plasma concentrations of immunoreactive or biologically active FSH and, to a lesser extent, of immunoreactive LH, relative to non-carriers (++). Bovine cDNA probes for the alpha gonadotrophin, FSH beta and LH beta genes were used to investigate FecB-specific differences in the mRNA species for the gonadotrophin subunits in pituitaries obtained from ++ and BB mid-luteal phase ovary-intact ewes, ovariectomized ewes and ovary-intact or ovariectomized ewes with hypothalamic-pituitary disconnection (HPD) given the same regimen of pulsatile GnRH. No FecB-specific differences in the number or size of mRNA transcripts detected by northern blotting were noted for any of these genes. Densitometry of the northern blots revealed no significant FecB-specific differences in the relative amounts of mRNA encoding alpha gonadotrophin, FSH beta or LH beta in the pituitaries from any of the experimental groups of ++ and BB sheep. Furthermore, there were no significant FecB-specific differences in the pituitary content of FSH or LH in these animals, despite significantly higher plasma concentrations of FSH in the ovary-intact and ovariectomized HPD groups. These data show that whereas the FecB gene causes increased plasma concentrations of FSH, no consistent effects can be demonstrated on pituitary gonadotrophin content or on gonadotrophin subunit gene transcription, using northern analysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Tissue‐Specific variation in the length of the 5′ untranslated region of the βa‐inhibin mRNA in sheep

The 5' untranslated region (UTR) of beta A inhibin mRNA was compared in a variety of sheep tissues, using primer extension. Considerable variation in the length and number of 5' extended products were noted between tissues. Specific bands were noted in ovarian follicular RNA, which were also present in samples from corpora lutea, stroma, and placental cotyledon RNA. Other extended products were observed in RNA from corpora lutea, stroma, cotyledon, pituitary, bone marrow, frontal cortex, medial basal hypothalamus, adrenal, liver, and kidney, which were not present or weakly represented in follicular RNA. Additional tissue-specific bands were noted in testis and bone marrow RNA. No specific differences in the lengths of the 5' UTR of the beta A inhibin mRNA were observed in sheep homozygous for the Booroola fecundity gene FecB, in any tissue studied. The coding region of ovine beta A inhibin cDNA was sequenced and a genetic polymorphism confirmed within or close to the ovine beta A inhibin gene. We conclude that the beta A inhibin gene is expressed widely in the sheep. Furthermore there is variation in the length of the 5' UTR of beta A inhibin mRNA between male and female gonads and other tissues, implying that expression of this gene is differentially controlled. However, the FecB mutation does not affect mRNA splicing events or the initiation site used in ovarian transcription. The mechanism by which the FecB mutation influences the amounts of beta A inhibin mRNA, follicle-stimulating hormone (FSH) secretion and ovulation rate has still to be elucidated.

The ovine Booroola fecundity gene (FecB) is linked to markers from a region of human chromosome 4q.

The autosomal Booroola fecundity gene (FecB) mutation in sheep increases ovulation rate and litter size, with associated effects on ovarian physiology and hormone profiles. Analysis of segregation in twelve families (379 female progeny) identified linkage between the mutation, two microsatellite markers (OarAE101 and OarHH55, Zmax > 9.0) and epidermal growth factor (EGF) from human chromosome 4q25 (Zmax > 3.0). The marker OarAE101 was linked to secreted phosphoprotein 1 (SPP1, which maps to chromosome 4q21-23 in man) in the test pedigrees and independent families (Zmax > 9.7). The identification of linkage between the FecB mutation and markers from human chromosome 4q is an important step towards further understanding the control of ovulation rates in mammals.

Effect of castration on plasma concentrations of luteinizing hormone and follicle-stimulating hormone in adult Merino rams which were homozygous carriers or non-carriers of the Booroola fecundity gene

Before castration, the mean plasma concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) did not differ between FF and ++ Booroola rams. After castration, mean LH and FSH concentrations increased after 8 h, and for the next 14 days the rate of increase in FSH, but not LH, secretion was significantly faster in FF than in ++ rams (P less than 0.05). Mean FSH concentrations over this period were significantly higher in FF than in ++ rams (P less than 0.05). In both genotypes, the ranked FSH values did not significantly change their order over time, i.e. a significant within-ram effect was noted (P less than 0.05). Repeated-measures analysis of variance indicated a significant effect of genotype on mean FSH secretion (P less than 0.05) and a significant effect of sire in the FF (P less than 0.05), but not the ++ (P = 0.76), genotype. From Day 28 to Day 58 after castration, FSH and LH concentrations were variable and no overall increases in concentrations were observed. The mean concentrations of both hormones over this period were not related to genotype. There were no gene-specific differences in pulsatile LH secretion 14 weeks after castration. However, the mean LH, but not FSH, response to a bolus injection of 25 micrograms of gonadotrophin-releasing hormone (GnRH) was significantly higher in FF than in ++ rams (P less than 0.05) and this was not significantly affected by sire. These studies support the hypothesis that the F gene is expressed in adult rams, in terms of pituitary responsiveness to an injection of GnRH and to the removal of the testes, but it is not clear from this study whether the influence of sire is related to or independent of the apparent gene-specific differences.

Plasma LH, FSH and testosterone concentrations in adult rams which were homozygous carriers or non-carriers of the Booroola fecundity gene

No gene-specific differences were found with respect to LH or testosterone pulsatile secretion (over 12 h), or in 12 hourly mean FSH concentrations in adult Booroola FF and ++ rams. Also, no differences between genotypes in the LH response to an injection of testosterone propionate, the FSH response to an infusion of bovine follicular fluid, or the testosterone response to injections of PMSG were noted. However, during the phase of seasonal testicular development, mean testosterone pulse amplitude (over 12 h) and the FSH response to 25 micrograms GnRH were higher in FF than in ++ rams (P less than 0.05); there were also significant effects of sire (P less than 0.05 in FF genotype only) and litter size (P less than 0.05) on testosterone pulse amplitude and GnRH-stimulated FSH release, respectively. During the breeding season, mean LH, but not FSH, concentrations were higher in FF than in ++ rams, after an injection of 0.5 micrograms GnRH; LH release was not affected by sire or litter size (P greater than 0.05). Long-term studies revealed that the FF rams were born in significantly larger litters, they weighed significantly less than ++ rams (P less than 0.05), and that bodyweight was significantly correlated (P less than 0.05) with litter size. There were no differences in testis size, and testis size was not significantly correlated with bodyweight. There was a strong tendency (P = 0.056) for overall mean FSH concentrations, measured weekly for 9 months, to be highest more often in FF than in ++ rams.(ABSTRACT TRUNCATED AT 250 WORDS)