Studies on ovarian maturation of the shrimp, metapenaeus ensis.

• Other Authors: Lo, Ting Sze.
• Format: Others
• Language: English; Chinese
• Published: 2003
• Subjects: Metapenaeus; Shrimps > Reproduction; Ovaries
• Online Access: http://library.cuhk.edu.hk/record=b5896079; http://repository.lib.cuhk.edu.hk/en/item/cuhk-324395

Lo Ting Sze. === Thesis submitted in: December 2002. === Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. === Includes bibliographical references (leaves 145-170). === Abstracts in English and Chinese. === Declaration --- p.i === Abstract --- p.ii === Acknowledgements --- p.vii === List of Contents === Contents in Brief --- p.viii === Contents in Detail --- p.ix === List of Figures and Tables === List of Figures --- p.xiii === List of Tables --- p.xvi === Chapter Chapter 1 --- Introduction and literature review General introduction --- p.1 === Chapter 1.1 --- Structural changes in ovary of penaeid shrimp during maturation --- p.3 === Chapter 1.2 --- Biochemical and physiological changes in ovary of penaeid shrimp during maturation --- p.7 === Chapter 1.2.1 --- Variation of protein content --- p.7 === Chapter 1.2.1.1 --- Changes of proteins in ovary of penaeid shrimp during maturation --- p.7 === Chapter 1.2.1.2 --- Biochemical characterization of vitellin --- p.8 === Chapter 1.2.1.3 --- Site of vitellogenin synthesis --- p.10 === Chapter 1.2.2 --- Variation of lipid composition --- p.12 === Chapter 1.2.2.1 --- Role of lipids in ovary of penaeid shrimp --- p.12 === Chapter 1.2.2.2 --- Changes of lipids in ovary of penaeid shrimp during maturation --- p.13 === Chapter 1.2.3 --- Variation of other nutrient contents --- p.16 === Chapter 1.3 --- Endocrine control of ovarian maturation in penaeid shrimp --- p.16 === Chapter 1.3.1 --- Gonad-inhibiting hormone (GIH) --- p.17 === Chapter 1.3.2 --- Crustacean hyperglycemic hormone (CHH) --- p.19 === Chapter 1.3.3 --- X-organ sinus gland complex (XOSG) and CHH neuropeptide family --- p.20 === Chapter 1.3.4 --- Gonad-stimulating hormone (GSH) --- p.22 === Chapter 1.3.5 --- Methyl farnesoate (MF) --- p.23 === Chapter 1.3.6 --- Neurotransmitters --- p.24 === Chapter 1.3.7 --- Other factors --- p.26 === Chapter 1.3.8 --- Androgenic hormone (AH) --- p.27 === Chapter 1.4 --- Ecology and reproductive biology of Metapenaeus ensis --- p.29 === Chapter 1.4.1 --- Identification --- p.29 === Chapter 1.4.2 --- Ecology --- p.29 === Chapter 1.4.3 --- Reproductive biology --- p.30 === Chapter 1.5 --- Objective of research --- p.32 === Chapter Chapter 2 --- "Variation of circulating gonad-inhibiting hormone (GIH) level during ovarian maturation in the shrimp, Metapenaeus ensis" === Chapter 2.1 --- Introduction --- p.34 === Chapter 2.2 --- Materials and methods --- p.35 === Chapter 2.2.1 --- Experimental animals and serum extraction --- p.35 === Chapter 2.2.2 --- Protein content assay --- p.36 === Chapter 2.2.3 --- SDS-polyacrymide gel electrophoresis (SDS-PAGE) --- p.39 === Chapter 2.2.4 --- Western blot --- p.40 === Chapter 2.2.5 --- Purification of IgG from antiserum --- p.42 === Chapter 2.2.6 --- ELISA assay (indirect ELISA) --- p.42 === Chapter 2.3 --- Results --- p.44 === Chapter 2.3.1 --- Experimental animals --- p.44 === Chapter 2.3.2 --- Purification of anti-GIH IgG --- p.46 === Chapter 2.3.3 --- Optimization of indirect ELISA --- p.46 === Chapter 2.3.4 --- Determination of circulating GIH in haemolymph --- p.51 === Chapter 2.4 --- Discussion --- p.55 === Chapter Chapter 3 --- "Identification of genes differentially expressed during ovarian maturation in the shrimp, Metapenaeus ensis" === Chapter 3.1 --- Introduction --- p.60 === Chapter 3.2 --- Materials and methods --- p.61 === Chapter 3.2.1 --- Outline of methodology --- p.61 === Chapter 3.2.2 --- Experimental animals --- p.62 === Chapter 3.2.3 --- Total RNA extraction --- p.62 === Chapter 3.2.4 --- RNA arbitrarily primed PCR (RAP-PCR) --- p.64 === Chapter 3.2.5 --- DNA electrophoresis by agarose gel --- p.68 === Chapter 3.2.6 --- DNA electrophoresis by polyacrylamide gel --- p.68 === Chapter 3.2.7 --- Preparation of DIG-labeled probe from PCR product --- p.69 === Chapter 3.2.8 --- Checking for DIG-labeling yield --- p.69 === Chapter 3.2.9 --- Excision of cDNA library from ovary of Metapenaeus ensis --- p.70 === Chapter 3.2.10 --- PCR screening of insertion sequence --- p.73 === Chapter 3.2.11 --- Dot-blot --- p.75 === Chapter 3.2.12 --- Dot-blot hybridization --- p.76 === Chapter 3.2.13 --- Growth of cell for plasmid preparation --- p.79 === Chapter 3.2.14 --- Permanent culture preparation --- p.80 === Chapter 3.2.15 --- DNA Sequencing --- p.80 === Chapter 3.2.16 --- RNA formaldehyde denaturing gel electrophoresis --- p.82 === Chapter 3.2.17 --- Northern blot --- p.84 === Chapter 3.2.18 --- Northern hybridization --- p.84 === Chapter 3.2.19 --- 5´ة RACE (rapid amplification of cDNA ends) --- p.88 === Chapter 3.3 --- Results --- p.92 === Chapter 3.3.1 --- Experimental animals --- p.92 === Chapter 3.3.2 --- Total RNA extraction --- p.92 === Chapter 3.3.3 --- RNA arbitrarily primed PCR (RAP-PCR) --- p.92 === Chapter 3.3.4 --- Titer of cDNA library from Metapenaeus ensis ovary --- p.95 === Chapter 3.3.5 --- PCR screening of insertion sequence --- p.95 === Chapter 3.3.6 --- Dot-blot hybridization --- p.99 === Chapter 3.3.7 --- DNA sequencing of differentially expressed genes --- p.96 === Chapter 3.3.8 --- Northern analysis --- p.99 === Chapter 3.3.8.1 --- Housekeeping gene - elongation factor 1α --- p.106 === Chapter 3.3.8.2 --- Translationally controlled tumor protein (TCTP) --- p.106 === Chapter 3.3.8.3 --- Cytoskeletal actin --- p.109 === Chapter 3.3.8.4 --- Keratin --- p.109 === Chapter 3.3.8.5 --- Heat shock cognate 70 kD protein (hsc70) --- p.109 === Chapter 3.3.8.6 --- Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) --- p.117 === Chapter 3.3.8.7 --- Arginine kinase --- p.117 === Chapter 3.3.8.8 --- High mobility group 1-like protein (HMGl-like protein) --- p.122 === Chapter 3.3.8.9 --- Nucleoside diphosphate kinase --- p.122 === Chapter 3.4 --- Discussion --- p.122 === Chapter 3.4.1 --- RAP-PCR and dot-blot hybridization --- p.122 === Chapter 3.4.2 --- Functions of differentially expressed genes in ovarian maturation --- p.128 === Chapter 3.4.2.1 --- Translationally controlled tumor protein (TCTP) --- p.128 === Chapter 3.4.2.2 --- Cytoskeletal actin --- p.130 === Chapter 3.4.2.3 --- Keratin --- p.130 === Chapter 3.4.2.4 --- Heat shock cognate 70 kD protein (hsc70) --- p.131 === Chapter 3.4.2.5 --- Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) --- p.134 === Chapter 3.4.2.6 --- Arginine kinase --- p.135 === Chapter 3.4.2.7 --- High mobility group 1-like protein (HMGl-like protein) --- p.136 === Chapter 3.4.2.8 --- Nucleoside diphosphate kinase --- p.137 === Chapter 3.4.3 --- Speculation on the molecular changes in ooctyes during ovarian maturation --- p.139 === Chapter Chapter 4 --- General conclusion --- p.142 === References --- p.145