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Endocrine and molecular regulation of ovarian antral follicular wave emergence and growth in sheep

dc.contributor.advisorRawlings, Normanen_US
dc.contributor.committeeMemberSingh, Baljiten_US
dc.contributor.committeeMemberHiebert, Lindaen_US
dc.contributor.committeeMemberHonaramooz, Alien_US
dc.contributor.committeeMemberKrone, Paten_US
dc.contributor.committeeMemberAmbrose, Divkaren_US
dc.creatorSeekallu, Srinivasen_US
dc.date.accessioned2009-10-19T17:12:59Zen_US
dc.date.accessioned2013-01-04T05:01:35Z
dc.date.available2010-10-21T08:00:00Zen_US
dc.date.available2013-01-04T05:01:35Z
dc.date.created2009-10en_US
dc.date.issued2009-10en_US
dc.date.submittedOctober 2009en_US
dc.description.abstractIn sheep, large ovarian antral follicles grow in waves with a periodicity of every 4 to 5 days; each wave is initiated by a peak in serum concentrations of follicle stimulating hormone (FSH). In the present thesis, follicular data and hormone estimations acquired from daily ultrasonography and blood samples, respectively, were used to study mechanisms regulating the number of follicular waves per estrous cycle. Using additional approaches such as implants releasing estradiol-17β and or progesterone, immunization against gonadotropin releasing hormone (GnRH), and injections of GnRH, the role of pulsed luteinizing hormone (LH) secretion and FSH peaks in follicular wave emergence and growth and the dependency of FSH peaks on pulsed GnRH secretion, were studied in sheep. The viability of aged follicles was also addressed. The results of the present studies showed that ewes with three or four waves per cycle had cycles of the same length. The inter-wave interval was longer for the first and the last or ovulatory wave of the cycle in three compared to four wave cycles. The length of the lifespan and regression phase of the largest follicle of a wave declined across the cycle as FSH peak concentration and amplitude decreased. The maximum follicular diameter of the largest follicle growing in the first wave and the last or ovulatory wave of the cycle was greater compared to other waves of the cycle. Treatment of anestrous ewes with estradiol releasing implants alone completely abolished pulsed LH secretion and suppressed follicular wave development; however, FSH secretion was only minimally affected and the pool of small follicles was not affected. When pulsed secretion of LH was restored by frequent injections of GnRH, follicular waves were re-established. Treatment of anestrous ewes with implants releasing estradiol and progesterone, decreased FSH peak amplitude and abolished LH pulses and follicular waves; the size of the pool of small follicles increased. Immunization against GnRH in anestrous ewes abolished pulsatile LH secretion and suppressed follicular wave emergence; however, FSH peaks continued to occur for several weeks. In cyclic ewes, creating an LH pulse frequency typical of the follicular phase, during the luteal phase of the cycle by giving GnRH, increased maximum diameter of the largest follicle in a wave and serum concentrations of estradiol and progesterone. The enhanced growth of follicles in a wave blocked the next expected FSH peak and its associated follicular wave. Decreasing LH pulse frequencies lower than the minimal frequency seen in the luteal phase, by implants releasing progesterone, did not affect the growth of follicular waves. It was previously demonstrated that treatment of non-prolific WWF ewes with Prostaglandin F₂ₐ (PGF₂ₐ) and medroxy progesterone acetate (MPA) increased the ovulation rate by adding ovulations from the penultimate wave in addition to the final wave of the cycle; however, fertility was not improved. In the last study of my thesis, we collected follicles, with an extended lifespan, from the penultimate wave of the cycle in ewes given the PGF₂ₐ and MPA treatment. We compared their quality with follicles from the final wave of the cycle by looking at the expression of markers of follicular development. The results showed that theca cells of follicles from the final wave had significantly higher mRNA expression for vascular endothelial growth factor (VEGF) compared to follicles from the penultimate wave. Granulosa cells of follicles from the final wave had significantly higher mRNA expression for connexion 43 (Cx43) compared to follicles from the penultimate wave. Protein expression for Cx43, proliferating cell nuclear antigen (PCNA) and Factor VIII was greater in follicles from the final compared to the penultimate wave. We concluded from the present studies that: 1) the mechanism that makes a three wave or four wave cycle is unclear; 2) some level of pulsatile LH secretion is required for an FSH peak to trigger emergence of follicular waves in anestrous ewes; 3) progesterone enhances the inhibitory effects of estradiol on FSH secretion in anestrous ewes, suppressing specifically FSH peak amplitude; 4) an endogenous rhythm may exist that drives the peaks in FSH secretion independent of secretory products from the follicles growing in a wave and pulsed GnRH secretion; 5) follicular waves in ewes, when exposed to an LH pulse frequency similar to the follicular phase, during the luteal phase of the cycle, when serum progesterone concentrations are high, can grow and function like ovulatory follicles growing in the follicular phase of the cycle; 6) expression of some markers of vascularization/ angiogenesis, gap-junctional communication and cell proliferation, appeared to be decreased in follicles from the penultimate compared to the final wave of an estrous cycle, when the lifespan of follicles from the penultimate wave was extended such that they were present in the ovary with follicles from the final wave of the cycle.en_US
dc.identifier.urihttp://hdl.handle.net/10388/etd-10192009-171259en_US
dc.language.isoen_USen_US
dc.subjectPenultimate waveen_US
dc.subjectFinal waveen_US
dc.subjectFactor VIIIen_US
dc.subjectProliferating cell nuclear antigenen_US
dc.subjectConnexin 43en_US
dc.subjectEndothelial nitric oxide synthaseen_US
dc.subjectVascular endothelial growth factoren_US
dc.subjectProgesteroneen_US
dc.subjectEstradiolen_US
dc.subjectOvaryen_US
dc.subjectFollicular Wavesen_US
dc.subjectSheepen_US
dc.subjectFollicle-stimulating hormoneen_US
dc.subjectLuteinizing hormoneen_US
dc.subjectThree wave cyclesen_US
dc.subjectFour wave cyclesen_US
dc.titleEndocrine and molecular regulation of ovarian antral follicular wave emergence and growth in sheepen_US
dc.type.genreThesisen_US
dc.type.materialtexten_US
thesis.degree.departmentVeterinary Biomedical Sciencesen_US
thesis.degree.disciplineVeterinary Biomedical Sciencesen_US
thesis.degree.grantorUniversity of Saskatchewanen_US
thesis.degree.levelDoctoralen_US
thesis.degree.nameDoctor of Philosophy (Ph.D.)en_US

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