The shift to group-housing of the breeding herd makes it difficult to manage animals individually. To maximize sow longevity and reproductive performance, gilts must be bred on second heat weighing between 135 and 160 kg. As gilts are typically fed ad libitum (AD), they often exceed targeted breeding weights, leading to structural and other problems resulting in culling at early parities. Thus, utilization of an electronic sow feeder (ESF) enables caretakers to manage animals individually in group-housing situations, by allotting predetermined amounts of feed. The specific objectives of this study were to: 1) to determine the effects of AD or restricted (RS) feeding bases (FB) on growth and sexual development in replacement gilts using an ESF; 2) to determine the growth and reproductive responses to gonadotropins in gilts fed on either AD or RS feeding bases using an ESF; and 3) to ascertain the effects of short-term increases in feed allowances on growth and sexual development in previously feed-restricted gilts using an ESF. Gilts employed in all studies were acclimated to an ESF (ACCUTEAM, Osborne Industries, Osborne, KS) beginning at 150 d of age, during a 10-d training period. Across all experiments, treatments were assigned at 160 d of age. In experiments 1 and 2, gilts received feed on either AD (5.00 kg/d) or RS (2.72 kg/d) bases. In the first study, estrus detection began at 160 d of age. In experiment 2, gilts assigned to receive gonadotropins were treated at 170 d of age, with estrus detection beginning the next d. Gilts fed on the RS basis were observed to have improved feed conversion efficiency compared to AD-fed gilts in experiments 1 and 2 (both P < 0.01); and flushed gilts in experiment 3 had G: F similar to RS gilts. Average daily gain was not affected by FB in the first and third experiments, however AD-fed gilts gained weight at a faster rate (P = 0.02) than RS gilts in experiment 2. Gilts fed AD consumed their allotments across more meals than those fed RS. Age at puberty onset was not affected by FB in any of the experiments. Further, P.G. 600 did not hasten the onset of puberty, irrespective of FB in experiment 2. In experiment 3, ovulation rate was numerically greatest for flushed gilts, and was significantly greater than RS gilts (P = 0.05). The ovulation rate for flushed gilts was similar to AD gilts. Across all experiments, feed disappearance was observed to decrease as AD-fed gilts approached estrus. In this series of experiments, FB did not alter puberty onset, however ovulation rate was increased when gilts were allocated short-term increases in feed allowances. Further, utilizing an ESF may augment current industry estrus detection methods by monitoring feeding behavior. / Doctor of Philosophy / On commercial swine operations in the U.S. sow removal rates due to death and culling exceed 40% annually. On average, sows are culled at parity 3 or 4, while at least this many parities are required before investment costs are recouped. Gilts are typically fed AD until BW of 135 kg are attained by approximately 200 d of age. Oftentimes, gilts fed on AD basis exceed targeted breeding weights, leading to structural and other problems resulting in culling at early parities. To improve sustainability and competitiveness of pork production systems, the industry must focus on proper gilt management practices to facilitate successful entry into the breeding herd and increase longevity and lifetime performance as sows. A series of 3 experiments were conducted to evaluate different feeding bases on growth and sexual development in gilts. The first experiment evaluated the impact of AD or RS feeding on growth and sexual development in gilts using an ESF. Age at puberty was not different in limit-fed gilts. Limit-fed gilts also grew at a similar rate accompanied with improved feed conversion compared to AD fed gilts. Results from this experiment indicate that gilts can be limit-fed prior during the period around puberty until entering the breeding herd without negative impacts to growth or sexual development. In the second experiment, the same feeding bases were utilized, but within feeding basis groups, gilts either received or did not receive treatment with exogenous gonadotropins. Age at puberty was similar, regardless of feeding basis or gonadotropin treatment. Feed conversion was improved in limit-fed gilts, however daily gain was improved for gilts receiving feed AD. Results from this experiment imply that limit feeding gilts has no negative impacts on sexual development, but daily gain is reduced. In the third experiment, previous feeding bases were used, and a third, nutritional flushing, was also studied, when gilts received AD feed 7 d after first estrus. Flushed gilts had feed conversion like that of limit-fed gilts with daily gain similar to that of AD gilts. Gilts which were flushed had ovulation rates which were similar to gilts receiving feed AD but was greater compared with limit-fed gilts. Results indicate that flush feeding can achieve daily gain similar to ad libitum feeding and feed conversion similar to limit-feeding, while improving ovulation rate compared to limit-feeding. Across all studies, feed disappearance decreased for AD fed gilts as they neared estrus. Overall, using an ESF allows for precise feeding of replacement gilts and allows producers to implement various feeding strategies across multiple animals on an individual basis. Monitoring feed behavior with an ESF may enhance current estrus detection practices.

Bibliography: leaves 230-294. / xiii, 294 leaves : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Analyses in detail a native small fragment of hCG/[beta] hCG subunit, the [beta]-core hCG fragment which is found in large amounts in urine and may be of clinical importance. The aims were to purify the fragment, analyse its protein and carbohydrate structure, develop direct assay methods for its measurement, and to examine it's distribution in body fluids and it's relationship with the intact hCG molecule during pregnancy. / Thesis (Ph.D.)--University of Adelaide, Dept. of Obstetrics and Gynaecology, 1993

Human chorionic gonodotropin (HCG) affected in various ways cell cultures infected with strains of Rous sarcoma virus (RSV). The cell cultures studied were chick embryo fibroblasts (CEF), normal rat kidney cells infected with temperature-sensitive mutant (LA31-NRK) and a wild type RSV (B77-NRK). HCG increased the rate of transformation and viral titer of CEF cells infected with RSV, but not B77-NRK. HCG increased significantly transformation rates of LA31-NRK, only if the temperature sensitive transformation process was first delayed by incubation at non-permissive temperatures. It is suggested that some postinfective, pretransformational event(s) may operate before viralmediated transformation rates are increased by HCG.

Orientador: Guilherme de Paula Nogueira / Banca: Ed Hoffmann Madureira / Banca: Ciro Moraes Barros / Resumo: Avaliou-se a secreção de LHe FSH de plasmático em novilhasdurantea exposição à diferentes concentrações de progesterona(P4) e após a administração deGnRH ou benzoatodeestradiol (BE). Novenovilhas Nelore, pré-sincronizadas com PgF2a, emintervalo de 12 dias, foram submetidas a 6protocolos comrepetições casualizadas. Os 3 grupos de P4foram: CL (Corpo Lúteo), IMPL+CL (DIB® ecorpo lúteo) e IMPL(DIB®), eapós a remoçãodaP4 estimuladascom GnRH ou BE. Durante a P4, foicoletado sangue a cada12h e nos dias 3, 4 e 5 acada 15 min durante 6 h em 1 animalde cada grupo, depois da retiradados implantes e/ou aplicação de PgF2a, coletas foramrealizadas a cada 3hpor 24 h (BE) ou a cadahora por10h (GnRH) para quantificação de LH e FSH. O exame ultrasonográfico foi realizado a cada 12 he após o términodascoletas de sangue às 24h e 48h. Às 12 h após colocação do implante, o grupo IMPL+CLapresentou menorconcentração de LHque o grupo IMPL,após 36 h os grupos IMPL+C e IMPL apresentaram menorconcentração de LH que o grupo CL e às 60 h o grupo IMPL apresentou menor secreção que o grupo CL. Após 24 h da colocação dos implantes, o grupo IMPL apresentou maior secreção de FSH que os demais grupos, e após 48 e 60 h o grupo IMPL+CL apresentou maior secreção de FSH que o grupo CL. No grupo IMPL, a amplitude máxima do pico de LH foi maior após o GnRH quando comparado com o BE. No grupo CL as novilhas apresentaram menor número de folículos ovarianos e maior diâmetro do maior folículo em relação aos grupos IMPL+CL e IMPL. Quando foi aplicado BE, o grupo IMPL+CL apresentou menor taxa de ovulação com 24 h que os grupos CL e IMPL. Os protocolos de sincronização da ovulação empregados foram ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The LH and FSH secretion and follicle profile was evaluated during expositionto different progesterone (P4) concentration and afterGnRH or estradiol benzoate (BE) administration inNelore heifers. Nine heifers were pré-sinchronized with PGF2a (two injection with 12 days interval) received 6 treatments randomly repeated . There were 3 P4 groups: CL (corpus luteum), IMPL+CL (DIB®and CL) and IMPL (DIB®)that were stimulatedwith GnRH or BE after P4 removal. During P4 blood samples were collected every12 hand on days 3, 4and 5 every 15 minfor 6 hfrom 1 animal per group, after P4 device removal, every 3 hfor 24 h (BE)orevery hour for 10 h(GnRH), for LH and FSH quantification. Ultrasound examination was realizedevery 12 h until the end of blood samples, andthereafter 24 and 48 h. At 12hafter implant insertion the IMPL+CLgroup presented lower LH concentration than IMPL group, after 36 hthe animals with implant presented lower LH concentration than CL group, and after 60h the IMPL group had lower LH secretionthan CL one. On IMPL group,themaximum LH peak amplitude was higher in animalsthat received GnRH comparedto BE. Group with CL presented fewer follicles and higher largest follicle diameterthan IMPL+CLand IMPL. When BE was injected the IMPL+CL group presented lower ovulation rateat 24 h than CL and IMPLgroups. The ovulation synchronization protocols used wereefficient in promoting a preovulatory LH peak in Neloreheifers, independentlyof an associated CL or not to progesterone device. The GnRH treatment induced a higher LH peak amplitude and with an higer efficiency in stimulating ovulation in 24h than BE treatment, in animals with higher progesterone...(Complete abstract, click electronic access below) / Mestre

Yam Kwan Mei. / Thesis submitted in: August 1999. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 108-129). / Abstracts in English and Chinese. / Abstract (in English) --- p.ii / Abstract (in Chinese) --- p.iv / Acknowledgement --- p.vi / Table of Contents --- p.ix / List of Tables --- p.xiv / List of Figures --- p.xv / Symbols and Abbreviations --- p.xviii / Scientific names --- p.xx / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1.1 --- Gonadotropins --- p.1 / Chapter 1.1.1 --- Structure --- p.1 / Chapter 1.1.2 --- Function --- p.3 / Chapter 1.1.3 --- Regulation --- p.9 / Chapter 1.1.3.1 --- GnRH --- p.9 / Chapter 1.1.3.2 --- Steroids --- p.11 / Chapter 1.1.3.3 --- Activin --- p.12 / Chapter 1.2 --- Activin Family of Growth Factors --- p.14 / Chapter 1.2.1 --- Structure --- p.14 / Chapter 1.2.2 --- Function --- p.17 / Chapter 1.3 --- Objectives --- p.22 / Chapter Chapter 2 --- Cloning of Goldfish Activin βA cDNA and the Expression of Its mRNA in Gonadal and Non-gonadal Tissues --- p.24 / Chapter 2.1 --- Introduction --- p.24 / Chapter 2.2 --- Materials and Methods --- p.25 / Chapter 2.2.1 --- Cloning of goldfish activin βA cDNA --- p.26 / Chapter 2.2.1.1 --- Cloning of the 5' and 3' cDNA ends --- p.26 / Chapter 2.2.1.2 --- Extension of the 5' and 3' fragments --- p.28 / Chapter 2.2.2 --- Sequencing of the cDNA --- p.28 / Chapter 2.2.2.1 --- Generation of pKS/GactβA constructs with insert in different orientations --- p.28 / Chapter 2.2.2.2 --- Generation of overlapping subclones --- p.29 / Chapter 2.2.2.3 --- Cycle sequencing --- p.30 / Chapter 2.2.2.4 --- Sequence analyses --- p.30 / Chapter 2.2.3 --- Isolation of total and messenger RNA --- p.30 / Chapter 2.2.3.1 --- Isolation of total RNA --- p.30 / Chapter 2.2.3.2 --- Isolation of messenger RNA --- p.31 / Chapter 2.2.4 --- Southern blot analysis --- p.32 / Chapter 2.2.5 --- Northern blot analysis --- p.33 / Chapter 2.2.6 --- Reverse transcription-polymerase chain reaction (RT-PCR) --- p.33 / Chapter 2.3 --- Results --- p.35 / Chapter 2.3.1 --- Cloning and sequence analysis of activin β A cDNA --- p.35 / Chapter 2.3.2 --- Distribution of activin βA mRNA in different tissues --- p.49 / Chapter 2.4 --- Discussion --- p.53 / Chapter Chapter 3 --- Establishment and Characterization of Stable Cell Lines for the Recombinant Production of Goldfish Activin A --- p.59 / Chapter 3.1 --- Introduction --- p.59 / Chapter 3.2 --- Materials and Methods --- p.60 / Chapter 3.2.1 --- Construction of expression plasmid --- p.60 / Chapter 3.2.2 --- Cell culture --- p.62 / Chapter 3.2.3 --- Transfection of CHO cells --- p.62 / Chapter 3.2.4 --- G418 selection of transfected CHO cells --- p.62 / Chapter 3.2.5 --- Activin bioassay (EDF-assay) --- p.63 / Chapter 3.2.6 --- Cloning of pBK/GactβA-transfected CHO cells by limited dilution --- p.63 / Chapter 3.2.7 --- Isolation of total RNA --- p.65 / Chapter 3.2.8 --- Northern blot analysis --- p.65 / Chapter 3.3 --- Results --- p.66 / Chapter 3.3.1 --- Optimization of G418 concentration for selection --- p.66 / Chapter 3.3.2 --- Expression of activin activity by pBK/GactpβA- transfected CHO cells --- p.67 / Chapter 3.3.3 --- Establishment and characterization of CHO cell lines that stably produce recombinant goldfish activin A --- p.67 / Chapter 3.4 --- Discussion --- p.73 / Chapter Chapter 4 --- Differential Regulation of Goldfish Gonadotropin (GTH-Iβ and GTH-IIβ) Gene Expression by Recombinant Goldfish Activin --- p.79 / Chapter 4.1 --- Introduction --- p.79 / Chapter 4.2 --- Materials and Methods --- p.82 / Chapter 4.2.1 --- Animals --- p.82 / Chapter 4.2.2 --- Drug treatment --- p.83 / Chapter 4.2.3 --- Primary culture of dispersed pituitary cells --- p.84 / Chapter 4.2.4 --- Southern blot analysis --- p.85 / Chapter 4.2.5 --- Isolation of total RNA --- p.86 / Chapter 4.2.6 --- Northern blot analysis --- p.86 / Chapter 4.2.7 --- Dot blot analysis --- p.87 / Chapter 4.2.8 --- Data analyses --- p.87 / Chapter 4.3 --- Results --- p.88 / Chapter 4.3.1 --- Probe specificity --- p.88 / Chapter 4.3.2 --- Effects of goldfish activin on pituitary GTH-Iβ and -IIβ mRNA expression --- p.88 / Chapter 4.3.3 --- Blockade of activin effects by follistatin --- p.92 / Chapter 4.4 --- Discussion --- p.96 / Chapter Chapter 5 --- General Discussion --- p.101 / Chapter 5.1 --- Overview --- p.101 / Chapter 5.2 --- Contribution of the Present Research --- p.103 / Chapter 5.2.1 --- Cloning of full-length goldfish activin βA cDNA --- p.103 / Chapter 5.2.2 --- Establishment of stable cell lines for the recombinant production of goldfish activin A --- p.104 / Chapter 5.2.3 --- Differential regulation of goldfish gonadotropin (GTH-Iβ and GTH-IIβ) gene expression by recombinant goldfish activin --- p.105 / Chapter 5.3 --- Future Research Direction --- p.107 / Chapter 5.3.1 --- Activin studies --- p.107 / Chapter 5.3.2 --- Gonadotropin studies --- p.107 / References --- p.108

Kwok Hin-Fai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 84-100). / Abstracts in English and Chinese. / Abstract (in English) --- p.i / Abstract (in Chinese) --- p.iv / Acknowledgement --- p.vi / Table of contents --- p.vii / List of figures --- p.xi / List of tables --- p.xiv / Symbols and abbreviations --- p.xv / List of fish names mentioned in the thesis --- p.xviii / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Gonadotropins / Chapter 1.1.1 --- Structure --- p.1 / Chapter 1.1.2 --- Function --- p.4 / Chapter 1.2 --- Gonadotropin receptor / Chapter 1.2.1 --- Structure --- p.5 / Chapter 1.2.2 --- Expression --- p.7 / Chapter 1.2.3 --- Signaling / Chapter 1.2.3.1 --- cAMP-mediated pathway --- p.7 / Chapter 1.2.3.2 --- Phospholipase C-mediated pathway --- p.9 / Chapter 1.2.4 --- Regulation of expression --- p.12 / Chapter 1.2.5 --- Desensitization of receptors / Chapter 1.2.5.1 --- Uncoupling --- p.13 / Chapter 1.2.5.2 --- Internalization --- p.13 / Chapter 1.3 --- Structure of ovarian follicles --- p.14 / Chapter 1.4 --- The project objectives and long-term significance --- p.16 / Chapter Chapter 2 --- Cloning and Characterization of Zebrafish Follicle-stimulating Hormone (FSH) and Luteinizing Hormone (LH) Receptors ´ؤ Evidence for Distinct Functions of FSH and LH in Follicle Development / Chapter 2.1 --- Introduction --- p.19 / Chapter 2.2 --- Materials and Methods / Chapter 2.2.1 --- Animals and chemicals --- p.22 / Chapter 2.2.2 --- Isolation of total RNA --- p.22 / Chapter 2.2.3 --- Cloning of zebrafish FSHR (zfFSHR) and LHR (zfLHR) cDNA fragments from the zebrafish ovary --- p.23 / Chapter 2.2.4 --- Rapid amplification of 5´ةcDNA ends (5'-RACE) and full-length cDNA --- p.24 / Chapter 2.2.5 --- Isolation of ovarian follicles --- p.25 / Chapter 2.2.6 --- Sampling of the ovaries from sexually immature zebrafish --- p.25 / Chapter 2.2.7 --- Reverse transcription-polymerase chain reaction (RT-PCR) --- p.25 / Chapter 2.2.8 --- Construction of expression plasmids --- p.26 / Chapter 2.2.9 --- Transient transfection and reporter gene assay --- p.27 / Chapter 2.2.10 --- Establishment and characterization of stable zfFSHR or zfLHR-expressing cell lines --- p.28 / Chapter 2.3 --- Results / Chapter 2.3.1 --- Cloning of FSHR and LHR cDNA from the zebrafish ovary --- p.29 / Chapter 2.3.2 --- Functional characterization of zfFSHR and zfLHR --- p.30 / Chapter 2.3.3 --- Expression of zfFSHR and zfLHR during sexual maturation --- p.31 / Chapter 2.3.4 --- Stage-dependent expression of zfFSHR and zfLHR in the ovarian follicles --- p.32 / Chapter 2.4 --- Discussion --- p.33 / Chapter Chapter 3 --- Down-regulation of FSHR and LHR Expression in the Zebrafish Follicle Ceils by Gonadotropin (hCG) and Its Sigaling Mechanism / Chapter 3.1 --- Introduction --- p.51 / Chapter 3.2 --- Materials and Methods / Chapter 3.2.1 --- Animals --- p.54 / Chapter 3.2.2 --- Chemicals and hormones --- p.54 / Chapter 3.2.3 --- Primary follicle cell culture --- p.55 / Chapter 3.2.4 --- Total RNA isolation --- p.55 / Chapter 3.2.5 --- "Validation of semi-quantitative RT-PCR assays for FSHR, LHR and GAPDH" --- p.56 / Chapter 3.2.6 --- Data analysis --- p.57 / Chapter 3.3 --- Results / Chapter 3.3.1 --- Validation of semi-quantitative RT-PCR assays --- p.57 / Chapter 3.3.2 --- Gonadotropin regulation of FSHR and LHR expression in cultured zebrafish ovarian follicle cells --- p.58 / Chapter 3.3.3 --- Effect of db-cAMP and forskolin on FSHR and LHR expression --- p.59 / Chapter 3.3.4 --- Effects of H89 on hCG-induced suppression of FSHR and LHR expression --- p.60 / Chapter 3.4 --- Discussion --- p.60 / Chapter Chapter 4 --- General Discussion --- p.75 / Chapter 4.1 --- Cloning of zebrafish FSHR and LHR cDNAs and demonstration of receptor specificity --- p.77 / Chapter 4.2 --- Evidence for the differential expression of FSHR and LHR in the zebrafish ovarian and follicle development --- p.78 / Chapter 4.3 --- Down-regulation of FSHR and LHR expression in the zebrafish follicle cells by gonadotropin (hCG) --- p.79 / Chapter 4.4 --- Future research direction --- p.80 / References --- p.84

So Wai-Kin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 100-127). / Abstracts in English and Chinese. / Acknowledgement --- p.I / Abstract (in English) --- p.II / Abstract (in Chinese) --- p.IV / Table of contents --- p.VI / List of Figures --- p.X / Symbols and Abbreviations --- p.XII / List of fish names mentioned in the thesis --- p.XIV / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Pituitary --- p.1 / Chapter 1.2 --- Gonadotropins --- p.1 / Chapter 1.2.1 --- Structure --- p.2 / Chapter 1.2.2 --- Signaling --- p.3 / Chapter 1.2.3 --- Expression --- p.5 / Chapter 1.2.4 --- Functions --- p.7 / Chapter 1.2.4.1 --- Gonadotropin actions on gametogenesis --- p.7 / Chapter 1.2.4.2 --- Gonadotropin actions on steroidogenesis --- p.8 / Chapter 1.2.5 --- Regulation --- p.9 / Chapter 1.2.5.1 --- Neuroendocrine control --- p.10 / Chapter 1.2.5.1.1 --- Gonadotropin-releasing hormone (GnRH) --- p.10 / Chapter 1.2.5.1.2 --- Dopamine (DA) --- p.12 / Chapter 1.2.5.2 --- Gonadal steroid feedback --- p.12 / Chapter 1.2.5.2.1 --- Positive feedback --- p.13 / Chapter 1.2.5.2.2 --- Negative feedback --- p.14 / Chapter 1.2.5.3 --- Paracrine regulators within pituitary --- p.15 / Chapter 1.3 --- Objectives of the present study --- p.16 / Chapter Chapter 2 --- "Molecular Cloning and Functional Characterization of Zebrafish FSHβ, LHβ and GTHα subunits" / Chapter 2.1 --- Introduction --- p.19 / Chapter 2.2 --- Materials and methods --- p.21 / Chapter 2.2.1 --- Chemicals --- p.21 / Chapter 2.2.2 --- Animals --- p.21 / Chapter 2.2.3 --- Genomic DNA isolation --- p.22 / Chapter 2.2.4 --- Total RNA isolation --- p.22 / Chapter 2.2.5 --- Cloning of zebrafish FSHp，LHβ and GTHa fragments --- p.23 / Chapter 2.2.5.1 --- LHβ and GTHα --- p.23 / Chapter 2.2.5.2 --- FSHβ --- p.23 / Chapter 2.2.6 --- "5'- and 3'-RACE of zebrafish FSHp, LHβ and GTHα subunits" --- p.24 / Chapter 2.2.7 --- Construction of expression constructs --- p.25 / Chapter 2.2.8 --- Cell culture and transfection of Flp-In´ёØ CHO cell --- p.26 / Chapter 2.2.9 --- Recombinant production of zebrafish FSH and LH --- p.27 / Chapter 2.2.10 --- Reverse transcription-polymerase chain reaction (RT-PCR) analysis --- p.27 / Chapter 2.2.11 --- Northern blot hybridization --- p.28 / Chapter 2.2.12 --- SEAP reporter gene assay --- p.28 / Chapter 2.2.13 --- Data analysis --- p.29 / Chapter 2.3 --- Results --- p.30 / Chapter 2.3.1 --- "Cloning of zebrafish FSHβ, LHβ and GTHα subunits" --- p.30 / Chapter 2.3.2 --- "Expression of zebrafish FSHp, LHβ and GTHα in the zebrafish pituitary" --- p.31 / Chapter 2.3.3 --- Recombinant production of zebrafish FSH and LH --- p.32 / Chapter 2.3.4 --- Functional analysis of zebrafish FSH and LH --- p.33 / Chapter 2.4 --- Discussion --- p.34 / Chapter Chapter 3 --- "Spatial Expression Patterns of Zebrafish FSHβ, LHβ and GTHα Subunits in the Pituitary and Their Temporal Expression Profiles during Sexual Maturation and Ovulatory Cycle" / Chapter 3.1 --- Introduction --- p.58 / Chapter 3.2 --- Materials and methods --- p.61 / Chapter 3.2.1 --- Chemicals --- p.61 / Chapter 3.2.2. --- Animals --- p.62 / Chapter 3.2.3 --- Total RNA isolation from zebrafish pituitaries and reverse transcription --- p.62 / Chapter 3.2.4 --- Validation of RT-PCR on single pituitary --- p.63 / Chapter 3.2.5 --- Real-time PCR --- p.64 / Chapter 3.2.6 --- Tissue preparation for in situ hybridization --- p.64 / Chapter 3.2.7 --- In situ hybridization --- p.65 / Chapter 3.2.8 --- Data analysis --- p.66 / Chapter 3.3 --- Results --- p.66 / Chapter 3.3.1 --- "PCR amplification of FSHβ, LHβ and GTHα and GAPDH in single zebrafish pituitary" --- p.67 / Chapter 3.3.2 --- "Establishement of real-time RT-PCR for zebrafish FSHβ, LHβ and GTHa and GAPDH" --- p.67 / Chapter 3.3.3 --- "Temporal expression profiles of zebrafish FSHβ, LHβ and GTHα subunits during sexual maturation" --- p.67 / Chapter 3.3.4 --- "Temporal expression profiles of zebrafish FSHp, LHβ and GTHα subunits during ovulatory cycle" --- p.68 / Chapter 3.3.5 --- "In situ hybridization of zebrafish FSHβ, LHβ and GTHα" --- p.69 / Chapter 3.4 --- Discussion --- p.70 / Chapter Chapter 4 --- General Discussion / Chapter 4.1 --- Cloning of zebrafish gonadotropin subunit cDNAs --- p.91 / Chapter 4.2 --- Bioactivity and receptor specificity of recombinant zebrafish FSH and LH --- p.91 / Chapter 4.3 --- Expression of gonadotropin subunits during zebrafish sexual maturation and ovulatory cycle --- p.92 / Chapter 4.4 --- "Localization of FSHβ, LHβ and GTHα subunits in zebrafish pituitary" --- p.93 / Chapter 4.5 --- Contributions of the present study --- p.94 / Chapter 4.6 --- Future prospects --- p.95 / References --- p.100

A series of experiments was conducted to evaluate the effects of nutrition during calfhood (defined as the period from 10 to 26-30 wk of age) and peripubertal period (defined as the period from 27-31 to 70-74 wk of age) on sexual development and reproductive function in beef bulls. The overall objective of these experiments was to evaluate the effects of nutrition on endogenous metabolic hormones (leptin, insulin, GH, and IGF-I), gonadotropins and testosterone concentrations, sexual development, sperm production, and semen quality in bulls. The results of these experiments demonstrated that nutrition affected GnRH secretion and sexual development in bulls. Increased nutrition during calfhood resulted in a more sustained increase in LH pulse frequency during the early gonadotropin rise and greater testicular development at maturity. On the other hand, low nutrition during calfhood suppressed LH secretion during the early gonadotropin rise and resulted in delayed puberty and reduced testicular development at maturity. When low nutrition was accomplished by restricted feed intake, hypothalamic and pituitary function were compromised and LH secretion was more severely affected. Temporal associations between LH secretion patterns and circulating IGF-I concentrations implied that IGF-I is a possible signal to the central metabolic sensor involved in translating body nutritional status to the GnRH pulse generator. Nutrition also affected testicular steroidogenesis (testosterone concentrations), indicating effects on the number or function of Leydig cells, or both. Age-related increases in physiological and GnRH-stimulated circulating testosterone concentrations were hastened in bulls receiving high nutrition and delayed in bulls receiving low nutrition; these effects were probably mediated by both LH secretion and IGF-I concentrations. Circulating leptin and insulin may have only permissive roles on GnRH secretion, but may enhance testicular development. Growth hormone concentrations decreased concomitantly with increasing IGF-I concentrations during sexual development in bulls, suggesting that the testes could contribute considerable amounts of circulating IGF-I. In conclusion, management strategies to optimize reproductive function in bulls should focus on increasing nutrition during calfhood.

A series of experiments was conducted to evaluate the effects of nutrition during calfhood (defined as the period from 10 to 26-30 wk of age) and peripubertal period (defined as the period from 27-31 to 70-74 wk of age) on sexual development and reproductive function in beef bulls. The overall objective of these experiments was to evaluate the effects of nutrition on endogenous metabolic hormones (leptin, insulin, GH, and IGF-I), gonadotropins and testosterone concentrations, sexual development, sperm production, and semen quality in bulls. The results of these experiments demonstrated that nutrition affected GnRH secretion and sexual development in bulls. Increased nutrition during calfhood resulted in a more sustained increase in LH pulse frequency during the early gonadotropin rise and greater testicular development at maturity. On the other hand, low nutrition during calfhood suppressed LH secretion during the early gonadotropin rise and resulted in delayed puberty and reduced testicular development at maturity. When low nutrition was accomplished by restricted feed intake, hypothalamic and pituitary function were compromised and LH secretion was more severely affected. Temporal associations between LH secretion patterns and circulating IGF-I concentrations implied that IGF-I is a possible signal to the central metabolic sensor involved in translating body nutritional status to the GnRH pulse generator. Nutrition also affected testicular steroidogenesis (testosterone concentrations), indicating effects on the number or function of Leydig cells, or both. Age-related increases in physiological and GnRH-stimulated circulating testosterone concentrations were hastened in bulls receiving high nutrition and delayed in bulls receiving low nutrition; these effects were probably mediated by both LH secretion and IGF-I concentrations. Circulating leptin and insulin may have only permissive roles on GnRH secretion, but may enhance testicular development. Growth hormone concentrations decreased concomitantly with increasing IGF-I concentrations during sexual development in bulls, suggesting that the testes could contribute considerable amounts of circulating IGF-I. In conclusion, management strategies to optimize reproductive function in bulls should focus on increasing nutrition during calfhood.

The present investigation was designed to further our understanding of the interaction of pituitary (luteinizing hormone, LH) and placental (chorionic gonadotropin, CG) gonadotropins with the primate corpus luteum. Studies were performed (1) to characterize the LH/CG receptor population in the