The Janus kinase (JAK) pathway is an integral part of signaling through a variety of ligands and receptors in mammals. The extensive reutilization and pleiotropy of this pathway in vertebrate development is conserved in other animals as well. In Drosophila melanogaster, JAK signaling is involved in embryonic pattern formation, sex determination, larval blood cell development, wing venation, planar polarity in the eye, and formation of other adult structures. Here we describe several roles for JAK signaling in Drosophila oogenesis. The gene for a JAK pathway ligand, unpaired, is expressed specifically in the polar follicle cells, two pairs of somatic cells at the anterior and posterior poles of the developing egg chamber. A primary defect of chambers with reduced JAK activity is fusion of successive chambers. These chambers exhibit an expansion of the polar cell population and concomitant loss of interfollicular stalk cells. Mosaic analysis of both JAK pathway transducers, hopscotch and stat92E, reveals that JAK signaling is specifically required in the somatic follicle cells. Another role of JAK signaling is in oocyte localization. In chambers mosaic for loss of hop activity, oocyte mislocalization results. Proper localization occurs only when the posterior follicle cells are wild type for hop.

The harvesting of immature mammalian oocytes (eggs) and their maturation in a laboratory environment, known as in-vitro maturation (IVM), provides an alternative to the harvesting of mature oocytes for in-vitro fertilisation (IVF) programs. The nutrient environment of an oocyte matured in vitro is known to have a significant effect on its potential to successfully mature, and it is desirable for the in-vitro nutrient environment to mimic the natural environment in vivo. This thesis describes an interaction between mathematical modelling and experimental investigation designed to build upon understanding of the nutrient environment of the oocyte in vivo, which is difficult to determine via experiment alone. A general mathematical model of nutrient transport to the oocyte, through its surrounding cumulus cells is developed. This model is applicable in-vivo and in-vitro across several species and to a number of important nutrients. Nutrient transport in this system - the cumulus-oocyte complex (COC) - is of particular importance, as it is this system that is normally removed for IVM treatments, and its solution under in-vivo conditions allows the nutrient concentration reaching the oocyte to be determined, given a known concentration immediately surrounding the COC. To successfully apply this model, parameters representing the rate of nutrient transport into cells within the COC must be accurately determined. These parameters are determined by a combination of experimental procedures and mathematical modelling in the case of an important nutrient to oocyte development, glucose. This work gives insight into the concentration dependence of glucose uptake into cell types that are important in regulating oocyte development, and to the behaviour of the oocyte itself with regard to glucose uptake. Finally models to describe the transport of two key nutrients, oxygen and glucose, from the vascular system in the ovary, through the ovarian follicle to the oocyte are developed. These make use of experimental results found in the study of glucose transport in the COC, and show that the geometry of the follicle has a significant impact on the nutrient environment of the COC, and hence by inference the nutrient environment of the oocyte. Work discussed in this thesis has been published [31, 156] and submitted for publication [30]. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1374636 / Thesis (Ph.D.) -- University of Adelaide, School of Mathematical Science, 2009

Thesis (Ph. D.)--University of Wisconsin--Madison, 1957. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 214-215).

Thesis (Ph. D.)--University of Hong Kong, 1999. / Includes bibliographical references (leaves 178-214).

Thesis (Ph. D.)--University of Wisconsin--Madison, 1981. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 125-131).

Thesis (M.Med.Sc.)--University of Hong Kong, 2003. / Also available in print.

Patients with dysfunctional ovaries, such as those with premature ovarian insufficiency and granulosa cell tumours, do not have normal follicle development and may not respond to traditional assisted reproductive techniques. Using the reaggregated ovary (RO) technique, these patients' oocytes may be reaggregated with functional supporting cells and cultured in vitro to develop fertilisable eggs. However, current research using ROs have only used murine ovaries as a somatic cell source. In this thesis, with the aim of moving towards a clinical treatment, we assessed follicle development in ROs in vitro and progressed to using the technique with human tissues. To assess whether an older murine somatic cell source resulted in advanced follicle development, and how follicle development differed between transplanted and cultured ROs, ROs were generated using postnatal day 2 (P2) and P6 mouse ovaries. To investigate theca cell development in follicles from cultured tissue, mouse ovaries were cultured with mouse serum or encapsulated in hyaluronan hydrogels. Prior to generating and culturing chimeric human-mouse ROs (HuMoROs), competent handling and digestion of bovine cortical tissue was required. Broadly, ROs generated from both P2 and P6 exhibited similar follicle development in vitro after 14 d of culture, and follicles from cultured ROs were more developed than those from transplanted ROs. Theca cell development observed in follicles from cultured ovaries was still poorer than those from in vivo ovaries, even when ovaries were cultured in mouse serum or encapsulated in a hyaluronan hydrogel. Finally, some follicles containing potential human oocytes developed within the generated HuMoROs after 7 d of culture. These results have highlighted the potential of the RO technique as a method to generate fertilisable eggs and identified further aspects which need to be targeted in order to improve the success of the technique.

Prolactin receptors (PRLr) were detected in anestrous (n=3), winter cycling (n=2), follicular (n=3) and luteal phase (n=3) equine ovaries by IHC. Follicle stages evaluated were primordial, preantral and antral. Receptors were detected in all follicle stages and in CL. PRLr staining was not different (P > 0.05) between primordial and preantral, but was greater (P < 0.001) in antral follicles. Primordial follicles stained weakest in anestrous and follicular phase ovaries, followed by luteal phase ovaries and was most intense in winter cycling. Staining in preantral follicles was weakest in anestrus, followed by follicular phase and highest in winter cycling and luteal phase. Staining was most intense in antral follicles with no difference (P > 0.05) between any of the reproductive states. Oocytes and ovulation fossa also possessed PRLr. In conclusion, concentrations of PRLr are highest in large, antral follicles suggesting a mechanistic role for PRL around the time of ovulation.

Establishment and maintenance of pregnancy is a central concern to the cattle industry, as it strongly impacts efficiency and profitability of beef cow-calf operations. The objective of this study was to determine if nutrient restriction impacts ovulatory follicle size and corpus luteum (CL) development and function of Bos taurus cows enrolled in estrous synchronization. A total of 26 Angus cows were housed in a facility equipped with a Calan gate system for individual animal intake. Cows were stratified by body weight (BW), and randomly assigned one of two nutritional treatments: 1) 100% of nutrient requirements (MTN; n=13) or 2) 70% of nutrient requirements (REST; n=13). Individual daily intakes were measured and adjusted weekly based on BW. Cattle underwent an acclimation period of 14 days and were exposed to nutritional treatments for 30 days prior to estrous synchronization. Body weight was measured daily using an automated scale and a conventional livestock scale at the beginning and end of the experiment. Cows were synchronized using a 7-day CO-synch + CIDR protocol beginning on day -10. Ultrasonography of the ovaries was performed at each event of the estrous synchronization protocol on days -10, -3, 0, 5, and 7. Blood samples were taken on days -10, -3, and daily from day 0 through 7 to observe changes in progesterone (P4). Data were analyzed as repeated measures using the MIXED procedure of SAS. Initial BW tended to differ between treatments (P = 0.07; MTN 597 ± 32 kg, REST 604 ± 32 kg), but MTN had greater final BW (P < 0.001; 687 ± 24 and 556 ± 27 kg, respectively) and greater average daily gain (1.35 ± 0.18 and -0.72 ± 0.21 kg/d, respectively, P < 0.001) than REST. Diameter of the largest follicle was similar (P = 0.851) between treatments at CIDR insertion (12.6 ± 0.6 mm) and CIDR removal (12.9 ± 0.4 mm) but was greater (P < 0.05) for MTN than REST cows at 60 hrs after CIDR removal (14.01 ± 0.6 and 12.37 ± 0.5 mm, respectively). Volume of CL was similar (P > 0.1) at 5 (3211 ± 113 mm3) and 7 (5280.3 ± 212 mm3) days after ovulation. Concentration of P4 did not differ on days -10, -3, or 0-5. However, on days 6 and 7, P4 was greater (P < 0.05) for MTN than REST (2.07 ± 0.15 and 1.65 ± 0.15, and 2.27 ± 0.15 and 1.83 ± 0.15 ng/mL, respectively). In conclusion, nutrient restriction to 70% of maintenance during estrous synchronization negatively affects diameter of the ovulatory follicle and circulating P4, but it did not affect CL volume in multiparous Bos taurus beef cows. / Master of Science / Ensuring successful pregnancy in beef cow-calf operations is crucial for the efficiency and profitability of the cattle industry. This study investigates the effects of nutrient restriction on ovulatory follicle size and corpus luteum (CL) volume in Angus cows undergoing estrous synchronization. A total of 26 cows were subjected to either a maintenance diet meeting 100% of nutrient requirements (MTN) or a diet providing 70% of nutrient requirements (REST). Intakes were updated weekly using computer software. The cows underwent a 30-day nutritional treatment before synchronization of ovulation. Results revealed that cows on the maintenance diet exhibited greater final body weight and average daily gain compared to those on the restricted diet. While estrus expression showed a numerical increase in MTN cows, the impact was not statistically significant. Analysis of ovulatory follicle size demonstrated that MTN cows had larger follicles 60 hours after synchronization compared to REST cows. Surprisingly, corpus luteum volume did not differ between the two groups at 5 and 7 days after ovulation. Additionally, circulating progesterone (P4) levels were affected by nutrient restriction, with notable differences observed on days 6 and 7. In summary, nutrient restriction during ovulation synchronization negatively influenced ovulatory follicle size and P4 levels, but did not affect corpus luteum volume in mature Angus cows. These findings contribute valuable insights for the cattle industry, emphasizing the importance of proper nutrition for optimal reproductive health in beef cows.

Ultrasound-guided transvaginal follicular aspiration was used to obtain oocytes from cows to study follicular development and oocyte morphology. Follicular aspiration was conducted once during wk 1 to 12 postpartum on 120 lactating cows with 6 groups, separated by biweekly intervals. Approximately one half of the aspirated cows at each session were from the early groups (wk 1-2, 3-4, or 5-6) and the other half from the later groups (wk 7-8, 9-10, or 11-12). On the day of aspiration the number of follicles on each ovary, and their sizes, small (2-5 mm), medium (6-10 mm) and large (≥ 11 mm), were recorded. The collected oocytes were morphologically classified into 4 grades, with 4 = excellent, 3 = good, 2 = fair, and 1 = poor. Blood samples from the jugular vein and follicular fluid samples from the largest follicle were collected in order to perform hormone and metabolite assays. Environmental data were obtained from the local airport. There was a significant (P < .01) quadratic days pre- and postpartum by parity interaction for BCS. Body condition score for older cattle was the lowest at 90 d prior to calving and changed the least amount over time, while youngest cattle had the highest initial BCS at d 90 prior to calving and had the greatest change in BCS over time. Body condition score was the highest during summer calving season (3.3 ± .06) compared to BCS during winter calving season (2.6 ± .06). But the loss in BCS was greater for cows that calved in summer (-0.53 ± .06) compared to cows that calved in winter (-0.07 ± .08). Increased serum NEFA concentrations with simultaneous decreases in serum insulin concentrations for younger cattle implied a more negative EB status than for older cattle. The total number of follicles and total number of oocytes retrieved was significantly (P < .001) affected by a linear days postpartum by parity interaction with younger cattle having linear increases compared to decreases in the total number of follicles for older cattle. Oocyte quality score was affected by the quadratic days postpartum by parity interaction (P < .01) and calving season (P < .01). Younger cattle had higher initial quality scores compared to older cattle, but older cattle had higher quality oocytes towards the end of the 12 wk period compared with younger cattle. Younger cattle had higher E2 and IGF-I concentrations in follicular fluid associated with a higher number of total follicles and number of oocytes, compared to older cattle. However, oocyte quality of younger cattle seemed to be reduced and oocytes were less competent than for older cattle. Cattle in 3rd and greater lactation showed very little change in BCS and hormone and metabolite measures during early lactation, with no apparent decrease in oocyte quality, despite the aging effect on follicle numbers. This study demonstrated that conditions related to early lactation have a negative effect on oocyte quality and endocrine measures of dairy cattle and that animals of different ages are differentially affected. / Master of Science