Luk Chun-yin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 119-131). / Abstracts in English and Chinese. / ABSTRACT --- p.i / ACKNOWLEDGEMENTS --- p.iv / LIST OF FIGURES --- p.x / LIST OF TABLES --- p.xii / Chapter CHAPTER 1 --- GENERAL INTRODUCTION --- p.1 / Chapter CHAPTER 2 --- LITERATURE REVIEW --- p.6 / Chapter 2.1 --- Introduction --- p.7 / Chapter 2.2 --- Ammonia chemistry --- p.10 / Chapter 2.3 --- Ammonia metabolism and excretion --- p.11 / Chapter 2.3.1 --- Ammonia production --- p.11 / Chapter 2.3.2 --- Blood levels of ammonia --- p.12 / Chapter 2.3.3 --- Ammonia Excretion --- p.17 / Chapter 2.4 --- Urea metabolism and excretion --- p.23 / Chapter 2.4.1 --- Urea Chemistry --- p.23 / Chapter 2.4.2 --- Urea production in fishes --- p.24 / Chapter 2.4.3 --- Argininolysis --- p.25 / Chapter 2.4.4 --- Uricolysis --- p.26 / Chapter 2.4.5 --- Ornithine-urea Cycle (OUC) --- p.28 / Chapter 2.4.5.1 --- Tilapia inhabiting the highly alkaline Lake Magadi --- p.32 / Chapter 2.4.5.2 --- High Ambient Ammonia --- p.33 / Chapter 2.4.5.3 --- Air Exposure --- p.34 / Chapter 2.4.5.4 --- Toadfishes --- p.34 / Chapter 2.4.6 --- Blood urea concentration --- p.35 / Chapter 2.4.7 --- Urea excretion in fishes --- p.37 / Chapter 2.4.7.1 --- Branchial urea excretion in fishes --- p.37 / Chapter 2.4.7.2 --- Mechanisms of renal excretion in fishes --- p.40 / Chapter 2.5 --- Influence of environmental salinity on nitrogen excretion in teleosts --- p.42 / Chapter CHAPTER 3 --- BODY COMPOSITION AND UREA BIOSYNTHESIS OF SPAR US SARBA IN DIFFERENT SALINITIES --- p.46 / Chapter 3.1 --- Introduction --- p.47 / Chapter 3.2 --- Materials and Methods --- p.49 / Chapter 3.2.1 --- Experimental animals --- p.49 / Chapter 3.2.2 --- Tissue sampling --- p.49 / Chapter 3.2.3 --- Water chemistry analysis --- p.50 / Chapter 3.2.4 --- Hematological parameters --- p.50 / Chapter 3.2.5 --- Metabolite and electrolyte contents --- p.51 / Chapter 3.2.6 --- Hepatic enzymes activities --- p.51 / Chapter 3.2.6.1 --- Tissue preparation --- p.51 / Chapter 3.2.6.2 --- Carbamyl phosphate synthetases (CPSases; E.C. 2.7.2.5) --- p.52 / Chapter 3.2.6.3 --- Ornithine carbamoyl transferase (OCTase; E.C. 2.1.3.3) --- p.53 / Chapter 3.2.6.4 --- Argininosuccinate synthetase (ASS; E.C. 6.3.4.5) --- p.54 / Chapter 3.2.6.5 --- Argininosuccinate lyase (ASL; E.C. 4.3.2.1) --- p.54 / Chapter 3.2.6.6 --- Arginase (ARG; 3.5.3.1) --- p.55 / Chapter 3.2.6.7 --- Glutamate dehydrogenase (EC 1.4.1.3) --- p.55 / Chapter 3.2.6.8 --- Uricase (E.C. 1.7.3.3) --- p.56 / Chapter 3.2.6.9 --- Allantoinase --- p.57 / Chapter 3.2.6.10 --- Allantoicase --- p.57 / Chapter 3.2.7 --- Statistical analysis --- p.58 / Chapter 3.3 --- Results --- p.59 / Chapter 3.3.1 --- "Changes in hepatosmatic index, renal somatic index, muscle water and lipid content and hematological parametersin response to different salinity acclimation" --- p.59 / Chapter 3.3.2 --- Changes in serum chemistry in response to different salinity acclimation --- p.60 / Chapter 3.3.3 --- Changes in hepatic ornithine-urea cycle enzyme activitiesin response to different salinity acclimation --- p.61 / Chapter 3.3.4 --- Changes in GDHase and uricolytic enzyme activitiesin response to different salinity acclimation --- p.62 / Chapter 3.4 --- Discussion --- p.71 / Chapter 3.4.1 --- Hematological responses --- p.72 / Chapter 3.4.2 --- Muscle moisture content --- p.74 / Chapter 3.4.3 --- Circulating electrolyte levels --- p.75 / Chapter 3.4.4 --- Circulating metabolites levels --- p.77 / Chapter 3.4.5 --- Urea metabolism --- p.80 / Chapter 3.4.5.1 --- Ornithine-urea cycle enzymes --- p.80 / Chapter 3.4.5.2 --- Carbamoyl phosphate synthetase isozymes --- p.81 / Chapter 3.4.5.3 --- Uricolytic pathway and argininolysis --- p.85 / Chapter 3.4.5.4 --- Influence of salinity on urea metabolism --- p.86 / Chapter 3.4.6 --- Conclusion --- p.87 / Chapter CHAPTER 4 --- EFFECT OF SALINITY ON NITROGEN EXCRETION OF SPARUS SARBA --- p.88 / Chapter 4.1 --- Introduction --- p.89 / Chapter 4.2 --- Materials and Methods --- p.91 / Chapter 4.2.1 --- Experimental animals --- p.91 / Chapter 4.2.2 --- Experimental protocol --- p.92 / Chapter 4.2.3 --- Determination of net ammonia and urea excretion rates --- p.94 / Chapter 4.2.4 --- Statistical analysis --- p.94 / Chapter 4.3 --- Results --- p.95 / Chapter 4.3.1 --- Net ammonia-N and urea-N excretion rates --- p.95 / Chapter 4.3.2 --- Changes in net ammonia-N and urea-N excretion ratesin response to abrupt hyposmotic exposure --- p.95 / Chapter 4.3.3 --- Changes in net ammonia-N and urea-N excretion rates after exposure to amiloride for 3 hours --- p.96 / Chapter 4.3.4 --- Changes in net urea-N excretion rates in response to elevated body urea levels --- p.96 / Chapter 4.3.5 --- Changes in net ammonia-N excretion rates in response to elevated body ammonia levels --- p.97 / Chapter 4.4 --- Discussion --- p.106 / Chapter 4.4.1 --- Influence of environmental salinity on net ammonia-N and urea-N excretion rates --- p.106 / Chapter 4.4.2 --- Effects of amiloride on nitrogen excretion --- p.109 / Chapter 4.4.3 --- Effect of increased body ammonia on ammonia excretion --- p.113 / Chapter 4.4.4 --- Changes in net urea-N excretion rates in response to elevated body urea levels --- p.113 / Chapter 4.5 --- Conclusion --- p.114 / Chapter CHAPTER 5 --- GENERAL CONCLUSION --- p.115 / references --- p.119

Hui Fong Fong Liza. / Thesis submitted in: December 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 130-182). / Abstracts in English and Chinese. / Chapter I --- Title page --- p.I / Chapter II --- Thesis committee --- p.II / Chapter III --- Acknowledgements --- p.III / Chapter IV --- Abstract (Chinese version) --- p.IV / Chapter V --- Abstract (English version) --- p.VII / Chapter VI --- Table of contents --- p.X / Chapter VII --- List of figures --- p.XIV / Chapter VIII --- List of table --- p.XVIII / Chapter Chapter 1: --- General introduction --- p.1 / Chapter Chapter 2: --- Literature review --- p.5 / Chapter 2.1. --- Gill --- p.5 / Chapter 2.2. --- Chloride cells (Mitochondria-rich cells) --- p.6 / Chapter 2.2.1. --- Ion extrusion by fish in seawater --- p.9 / Chapter 2.2.2. --- Ion uptake by fish in hypo-osmotic condition --- p.12 / Chapter 2.3. --- Sparus sarba (Silver seabream) --- p.14 / Chapter 2.4. --- Sodium-potassium adenosinetriphosphatase (Na+-K+-ATPase) --- p.15 / Chapter 2.4.1. --- Na+-K+-ATPase α-subunit --- p.17 / Chapter 2.4.2. --- Na+-K+-ATPase β-subunit --- p.18 / Chapter 2.4.3. --- Regulation of Na+-K+-ATPase --- p.20 / Chapter 2.5. --- Hormones --- p.21 / Chapter 2.5.1. --- Growth hormone-prolactin family --- p.21 / Chapter 2.5.2. --- Structure of hormones --- p.22 / Chapter 2.5.2.1. --- Structure of growth hormone and prolactin in fish --- p.22 / Chapter 2.5.2.2. --- Structure of insulin-like growth factors in fish --- p.26 / Chapter 2.5.2.3. --- Structure of Cortisol in fish --- p.27 / Chapter 2.5.3. --- Regulation of hormones --- p.28 / Chapter 2.5.3.1. --- Regulation of growth hormone in fish --- p.28 / Chapter 2.5.3.2. --- Regulation of prolactin in fish --- p.32 / Chapter 2.5.3.3. --- Regulation of insulin-like growth factor-I in fish --- p.33 / Chapter 2.5.3.4. --- Regulation of Cortisol in fish --- p.33 / Chapter 2.5.4. --- Functions of hormones --- p.33 / Chapter 2.5.4.1. --- Functions of growth hormone in fish --- p.33 / Chapter 2.5.4.2. --- Functions of prolactin in fish --- p.39 / Chapter 2.5.4.3. --- Functions of insulin-like growth factor-I in fish --- p.44 / Chapter 2.5.4.4. --- Functions of Cortisol in fish --- p.45 / Chapter 2.5.4.5. --- "Combined effects of GH, IGF-I, PRL and Cortisol" --- p.49 / Chapter 2.6. --- Salinity effects on Na+-K+-ATPase expression --- p.52 / Chapter Chapter 3: --- In vitro effect of hormones on branchial Na+-K+- ATPase expression in marine teleost Sparus sarba --- p.58 / Chapter 3.1. --- Abstract --- p.58 / Chapter 3.2. --- Introduction --- p.60 / Chapter 3.3. --- Materials and methods --- p.62 / Chapter 3.3.1. --- Overall experimental design --- p.62 / Chapter 3.3.2. --- Fish preparation --- p.62 / Chapter 3.3.3. --- Tissue sampling --- p.62 / Chapter 3.3.4. --- RNA extraction and dot blot analysis --- p.63 / Chapter 3.3.5. --- Protein extraction --- p.65 / Chapter 3.3.6. --- Protein quantification --- p.65 / Chapter 3.3.7. --- Na+-K+-ATPase activity --- p.65 / Chapter 3.3.8. --- Protein gel electrophoresis and immunoblotting (Western blotting) --- p.66 / Chapter 3.3.9. --- Statistical analysis --- p.67 / Chapter 3.4. --- Results --- p.69 / Chapter 3.4.1. --- Dot blot analysis of Na+-K+-ATPase mRNA subunits --- p.69 / Chapter 3.4.2. --- Analysis of Na+-K+-ATPase protein α-subunit --- p.81 / Chapter 3.4.3. --- Analysis of Na+-K+-ATPase activity --- p.87 / Chapter 3.5. --- Discussion --- p.92 / Chapter 3.5.1. --- Effects of rbGH and rbIGF-I on Na+-K+-ATPase expression --- p.92 / Chapter 3.5.2. --- Effects of oPRL on Na+-K+-ATPase expression --- p.102 / Chapter 3.5.3 --- Effects of Cortisol on Na+-K+-ATPase expression --- p.104 / Chapter 3.6. --- Conclusion --- p.108 / Chapter Chapter 4: --- In vivo effect of salinity on branchial Na+-K+-ATPase expression in marine teleost Sparus sarba --- p.109 / Chapter 4.1. --- Abstract --- p.109 / Chapter 4.2. --- Introduction --- p.110 / Chapter 4.3. --- Materials and methods --- p.112 / Chapter 4.3.1. --- Overall experimental design --- p.112 / Chapter 4.3.2. --- Fish preparation --- p.112 / Chapter 4.3.3. --- Tissue sampling --- p.113 / Chapter 4.3.4. --- "RNA extraction, dot blot analysis, protein extraction, quantification, Na+-K+-ATPase activity, protein gel electrophoresis and immunoblotting (Western blotting)" --- p.113 / Chapter 4.3.5. --- Statistical analysis --- p.114 / Chapter 4.4. --- Results --- p.114 / Chapter 4.4.1. --- Dot blot analysis of Na+-K+-ATPase mRNA subunits --- p.114 / Chapter 4.4.2. --- Analysis of Na+-K+-ATPase protein a-subunit --- p.114 / Chapter 4.4.3. --- Analysis of Na+-K+-ATPase activity --- p.115 / Chapter 4.5. --- Discussion --- p.120 / Chapter 4.6. --- Conclusion --- p.125 / Chapter Chapter 5: --- General discussion and conclusion --- p.126 / References --- p.130

Fung Chun Kit. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 117-139). / Abstracts in English and Chinese. / Declaration of Originality --- p.i / Acknowledgements --- p.ii / Abstract --- p.iii / 摘要 --- p.v / List of figures --- p.vi / List of tables --- p.viii / List of abbreviations --- p.ix / Chapter Chapter I --- General introduction --- p.1 / Chapter 1.1 --- Different fish habitats with various salinities --- p.1 / Chapter 1.2 --- Osmotic challenges faced by teleosts --- p.2 / Chapter 1.3 --- High ionic strength results in DNA damage and excess water gain --- p.3 / Chapter 1.4 --- Osmoregulatory organs and mechanisms for osmotic balance --- p.4 / Chapter 1.5 --- Different tolerance to salinity changes --- p.8 / Chapter 1.6 --- Effective communication among osmoregulatory organs --- p.9 / Chapter 1.7 --- Introduction to GH and PRL --- p.9 / Chapter 1.8 --- Structure of the GHR and PRLR --- p.10 / Chapter 1.9 --- Hypoosmoregulatory action of GH/IGF-I axis in teleosts --- p.11 / Chapter 1.10 --- Hyperosmoregulatory action of PRL in teleosts --- p.11 / Chapter Chapter II --- Research rationale --- p.13 / Chapter 2.1 --- Physiological importance of osmoregulation in fish --- p.13 / Chapter 2.1.1 --- Energy metabolism --- p.13 / Chapter 2.1.2 --- Growth --- p.14 / Chapter 2.1.3 --- Immunity --- p.14 / Chapter 2.1.4 --- Reproduction --- p.15 / Chapter 2.2 --- Aquaculture importance --- p.15 / Chapter 2.3 --- Unknown molecular regulatory mechanism of hormones during salinity changes in fish --- p.16 / Chapter 2.4 --- Animal model --- p.17 / Chapter Chapter III --- In vivo studies of sbGHR and sbPRLR expression in osmoregulatory organs in response to salinity changes --- p.18 / Chapter 3.1 --- Introduction --- p.18 / Chapter 3.1.1 --- Dynamic change of GH level during salinity changes --- p.18 / Chapter 3.1.2 --- Dynamic change of PRL level during salinity changes --- p.19 / Chapter 3.1.3 --- In vitro studies of GH and PRL release from teleost pituitary in response to extracellular osmolality changes --- p.20 / Chapter 3.1.4 --- Biological actions of GH and PRL through the GHR and PRLR --- p.21 / Chapter 3.2 --- Materials and methods --- p.23 / Chapter 3.3 --- Results --- p.28 / Chapter 3.4 --- Discussion --- p.36 / Chapter 3.4.1 --- Plasma osmolality change during salinity changes --- p.36 / Chapter 3.4.2 --- Gene expression after HSW exposure --- p.38 / Chapter 3.4.3 --- Ionic mediators of the gene expression --- p.43 / Chapter 3.4.4 --- Gene expression after BW exposure --- p.44 / Chapter 3.4.5 --- Dynamic changes of the GHR and PRLR in response to salinity changes --- p.45 / Chapter 3.4.6 --- Regulation of the gene expression in response to salinity changes --- p.46 / Chapter Chapter IV --- Gene expression of sbGHR in liver during salinity changes --- p.49 / Chapter 4.1 --- Introduction --- p.49 / Chapter 4.1.1 --- Responses of the somatotropic axis to salinity changes in fish --- p.49 / Chapter 4.2 --- Materials and methods --- p.52 / Chapter 4.3 --- Results --- p.56 / Chapter 4.4 --- Discussion --- p.60 / Chapter 4.4.1 --- Inhibition of GHR and IGF-I gene expression in liver during HSW exposure --- p.60 / Chapter 4.4.2 --- Downregulation of GHR gene expression by hyperosmotic stress --- p.62 / Chapter 4.4.3 --- Growth retardation of fish during hyperosmotic environment --- p.64 / Chapter Chapter V --- Gene expression studies of sbPRLR in gill organ culture --- p.68 / Chapter 5.1 --- Introduction --- p.68 / Chapter 5.1.1 --- Functions of gill in fish osmoregulation --- p.68 / Chapter 5.1.2 --- Gill culture as a model for osmoregulation studies --- p.69 / Chapter 5.2 --- Materials and methods --- p.70 / Chapter 5.3 --- Results --- p.71 / Chapter 5.4 --- Discussion --- p.73 / Chapter Chapter VI --- Regulation of gene expression of sbGHR in liver during hyperosmotic stress: promoter studies --- p.75 / Chapter 6.1 --- Introduction --- p.75 / Chapter 6.1.1 --- What is a promoter? --- p.75 / Chapter 6.1.2 --- Promoter studies of GHR gene --- p.76 / Chapter 6.2 --- Materials and methods --- p.78 / Chapter 6.3 --- Results --- p.85 / Chapter 6.4 --- Discussion --- p.104 / Chapter Chapter VII --- General discussion and future perspectives --- p.111 / References --- p.117

All these findings can help us to elucidate the mechanisms for the fish to detect changing osmotic conditions and transform signals to osmoregulatory responses. / In the first part of the study, PRL and PRL-releasing peptide (PrRP) cDNAs have been isolated from euryhaline silver sea bream. The PRL cDNA consists of 1360 bp encoding 212 amino acids whereas the PrRP cDNA contains 631 bp encoding prepro-PrRP with 122 amino acids. PRL mRNA was uniquely expressed in sea bream pituitary but PrRP mRNA was expressed in a variety of tissues. Expression levels of both PRL and PrRP mRNA have been examined in sea bream adapted to different salinities (0, 6, 12, 33 and 50 ppt). In pituitary, both PRL and PrRP mRNA were synchronized in their expression, being significantly higher in fish adapted to low salinities (0 and 6 ppt), but the expression profile of hypothalamic PrRP in different salinities was different. These data suggested that PrRP may possibly act as a local modulator in pituitary rather than a hypothalamic factor for regulating pituitary PRL expression in silver sea bream. / In the second part of the study, silver sea bream abruptly transferred from 33 to 6 ppt exhibited a remarkable pituitary PRL secretion following closely with the temporal changes in serum osmolality and ion levels. In order to investigate the direct effect of extracellular osmolality to pituitary PRL secretion, sea bream pituitary cells were dispersed and exposed to a medium with reduced ion levels and osmolality. PRL released from pituitary cells was found to be significantly elevated. In hyposmotic exposed anterior pituitary cells, cell volume exhibited a 20% increase when exposed to a medium with a 20% decrease in osmolality. These enlarged pituitary cells did not shrink until the surrounding hyposmotic medium was replaced, a phenomenon suggesting an osmosensing ability of silver sea bream PRL cells for PRL secretion in response to a change in extracellular osmotic pressure. / In the third part, olfactory rosette in the nasal cavity was surgically removed from silver sea bream adapted to 6 ppt and 33 ppt and mRNA expression of PRL and PrRP in silver sea bream were measured. The elevated pituitary PRL and PrRP mRNA expression levels as seen in 6 ppt-adapted fish were abolished by this olfactory lamellectomy. On the other hand, hypothalamic PrRP mRNA expression in 6 ppt-adapted fish did not change but those in 33 ppt-adapted fish increase significantly after olfactory lamellectomy. These data suggest a possible osmosensing role of the olfactory system for regulation of PRL expression during hypo-osmotic acclimation of the fish. Besides, calcium-sensing receptor (CaSR) was cloned and its mRNA expression in olfactory system, as shown in other fish species previously, was investigated. However, no CaSR expression could be detected in olfactory rosette and nerve but its expression was demonstrated in osmoregulatory tissues and brain. There was no significant difference in CaSR mRNA expression in pituitary, kidney and anterior intestine of fish adapted to different salinities. These studies could not provide conclusive evidence to correlate CaSR with osmosensing in silver sea bream. / The present study used silver sea bream (Sparus sarba ) as a euryhaline fish model to investigate the regulation of prolactin (PRL) expression and secretion in fish adapted to different salinities. / Kwong, Ka Yee. / Adviser: Norman Y. S. Woo. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3248. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 154-184). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Branchial carbonic anhydrase was purified from silver seabream (Sparus sarba) and antibody against the enzyme was obtained by immunization in rabbits. An assay for quantifying the activity of carbonic anhydrase was developed. Using enzymatic and immunological techniques, the activity, expression and distribution of branchial carbonic anhydrase of silver seabream acclimated to different salinities were studied. Fish gill is one of the most important organs involved in various homeostatic processes. The ability of euryhaline fish to maintain constant internal ionic balance is crucial for the survival of the fish upon change in salinity. The presence of carbonic anhydrase in the chloride cells was suggested to be an important enzyme involved in ion regulation of fish. / In the present study, branchial carbonic anhydrase and erythrocyte carbonic anhydrase were purified from the gill cells of silver seabream with p-aminomethylbenzenesulfonamide-agarose affinity column. They were predominantly cytosolic with a molecular size of 26.6 k Da for branchial carbonic anhydrase and 28.6 k Da for erythrocyte carbonic anhydrase. Investigation of kinetic properties towards the inhibitor acetazolamide has helped determine the inhibition constants (Ki of branchial carbonic anhydrase: 0.54 x 10-9; Ki of erythrocyte carbonic anhydrase: 0.22 x 10-9). The difference in molecular size and inhibition constant towards acetazolamide supported the view that branchial carbonic anhydrase and erythrocyte carbonic anhydrase were two different isozymes. Polyclonal antibody specific to seabream branchial carbonic anhydrase was obtained by immunization in rabbit. The distribution of branchial carbonic anhydrase in the gill of seabream acclimated to different salinities was studied with immunohistochemical method. The enzyme was mainly located at the interlamellar region. The effect of salinity (0, 6, 12, 33, 50 and 70 ‰) acclimation on the expression and activities of branchial carbonic anhydrase has shown a U-shape pattern from freshwater to double-strength seawater on the quantity of seabream branchial carbonic anhydrase. Higher amount of branchial carbonic anhydrase in freshwater was consistent with the current view that the enzyme was actively involved in the ion uptake process through the hydration of carbon dioxide to produce bicarbonate ion and proton for the exchange of chloride and sodium ions, respectively. An interesting finding was obtained with elevated amount of branchial carbonic anhydrase in seabream acclimated to double-strength seawater and the possible role of the enzyme in such extreme environment was discussed. / This study has provided useful information on the properties, localizations and activities of branchial carbonic anhydrase in silver seabream for the understanding of the involvement of the enzyme in salinity adaptation of silver seabream. / Ma, Wing Chi Joyce. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3250. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 127-151). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

In the second part of our study, cDNA of AQP-1 and pro-vasotocin were cloned from the silver sea bream. An AQP-1 full clone was isolated from kidney and intestine and it consists of 904 bp with an open reading frame of 774 bp. The deduced amino sequence of sea bream AQP-1 shares highest identity with AQP-1a of gilthead sea bream (97.7%) and AQP-1a of other fish species (83.6% to 95.8%), however, considerably low identity was found between the silver sea bream AQP-1 and AQP-1b of gilthead sea bream (56%). The silver sea bream AQP-1 possesses basic features of a functional aquaporin and AQP-1, which includes two channel-forming asparagine-proline-alanine (NPA) signature motifs, six transmembrane domains, residues of the pore-forming region and a potential mercurial inhibiting site (Cys-178). The water channel was ubiquitously expressed in gills, liver, intestine, rectum, kidney, heart, urinary bladder and blood cells. A partial fragment of pro-vasotocin was isolated from hypothalamus of silver sea bream and consists of 184 bp, including encoding regions for the processing and amidation signal, vasotocin hormone and part of the neurophysin. / Lastly, single doses of cortisol (50 microg/g tissue) or vasotocin (1 microg/g tissue) were administered to seawater-acclimated sea bream with further three-day stabilizing period in seawater followed by an abrupt 6‰ exposure or administered to seawater transfer controls for three days. Cortisol markedly stimulated intestinal expression of AQP-1 in both the seawater transfer control and abrupt 6‰ transfer groups. Vasotocin treatment did not significantly modify AQP-1 expression in all tested organs. Hypothalamic pro-vasotocin expression levels were similar among different treatment groups. / Semi-quantitative RT-PCR analysis was used for studying the effect of salinity and hormones on expression of AQP-1 and pro-vasotocin. In the long-term salinity acclimation experiment, the sea bream were acclimated to six different salinity regimes (0‰, 6‰, 12‰, 33‰, 50‰, 70‰) for four weeks. The abundance of AQP-1 transcript was the highest in intestine of 70‰-acclimated fish among different salinity groups and there was also a statistically significant increase in 12‰-acclimated fish. Branchial AQP-1 expression was significantly upregulated in sea bream acclimated to freshwater. In contrast, the hypothalamic pro-vasotocin expression was significantly downregulated during freshwater acclimation. In addition, the sea bream were also subjected to an abrupt 6%o transfer at different time intervals (2, 6, 12, 72 and 168 hours). RT-PCR analysis revealed there was a transient decrease in branchial AQP-1 expression two hours after abrupt hypo-osmotic exposure and the expression levels subsequently returned to the seawater control levels. The expression levels of hypothalamic pro-vasotocin were not significantly altered by the abrupt exposure treatment. / The present experiments investigated the effects of salinity and hormones on the relative expression of hypothalamic pro-vasotocin, and aquaporin-1 (AQP-1) in intestine, gills and kidney of the silver sea bream Sparus sarba. With the use of immunohistochemical techniques, immunoreactivity of AQP-1 was detected at the basal side of enterocytes and gill chloride cells, and at the apical brush border of kidney tubules whereas AQP-3 was only localized in similar positions in the gills and intestines. AQP-1 was relatively more ubiquitous than AQP-3 and was localized with same cell types as the electrogenic Na+-K+-ATPase in gills and kidney. / The present study had demonstrated the responsiveness of intestinal and branchial AQP-1 expressions of the silver sea bream to environmental salinity perturbations. Further to this, cortisol was observed to upregulate the transcription of AQP-1 in the intestine. Pro-vasotocin expression was altered by long-term salinity adaptation, however, the linkage of this alteration to AQP-1 functioning in different osmoregulatory organs is yet to be elucidated. / Luk, Chun Yin. / Adviser: Norman Y. S. Woo. / Source: Dissertation Abstracts International, Volume: 72-04, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 200-222). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Wong Kwok-Shing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 89-107). / Abstracts in English and Chinese. / Title --- p.i / Abstract (English) --- p.ii / Abstract (Chinese) --- p.v / Content --- p.vii / Acknowledgement --- p.x / Abbreviation --- p.xii / Lists of tables and figures --- p.xiii / Chapter Chapter 1 --- General introduction --- p.1 / Chapter Chapter 2 --- "Effects of salinity on the cardiovascular responses and dipsogenic behaviors and silver seabream, Sparus sarba." / Chapter 2.1 --- Literature review / Chapter 2.1.1 --- Teleost euryhalinity --- p.5 / Chapter 2.1.2 --- Salinity and blood respiratory properties --- p.7 / Chapter 2.1.3 --- Salinity and blood volume --- p.8 / Chapter 2.1.4 --- Salinity and blood pressure --- p.10 / Chapter 2.1.5 --- Intestine physiology --- p.12 / Chapter 2.1.6 --- Summary --- p.14 / Chapter 2.2 --- Materials and methods / Chapter 2.2.1 --- Experimental animals --- p.19 / Chapter 2.2.2 --- Salinity adaptation --- p.19 / Chapter 2.2.3 --- Drinking rate measurement --- p.19 / Chapter 2.2.4 --- Respiratory characteristics --- p.20 / Chapter 2.2.5 --- Blood volume measurement --- p.21 / Chapter 2.2.6 --- Blood pressure experiment --- p.23 / Chapter 2.2.7 --- Statistical analysis --- p.23 / Chapter 2.3 --- Results / Chapter 2.3.1 --- Drinking rate --- p.25 / Chapter 2.3.2 --- Oxygen dissociation curves --- p.27 / Chapter 2.3.3 --- Blood volume --- p.29 / Chapter 2.3.4 --- Blood pressure --- p.31 / Chapter 2.4 --- Discussion / Chapter 2.4.1 --- Drinking rate --- p.36 / Chapter 2.4.2 --- Oxygen dissociation curves --- p.37 / Chapter 2.4.3 --- Blood volume --- p.38 / Chapter 2.4.4 --- Blood pressure --- p.40 / Chapter Chapter 3 --- "Manipulation of renin-angiotensin system in relation to the cardiovascular responses and dipsogenic behaviors of silver seabream, Sparus sarba." / Chapter 3.1 --- Literature review / Chapter 3.1.1 --- Renin angiotensin system (RAS) --- p.41 / Chapter 3.1.2 --- RAS and blood pressure --- p.47 / Chapter 3.1.3 --- RAS and drinking --- p.53 / Chapter 3.1.4 --- RAS and Cortisol --- p.55 / Chapter 3.1.5 --- RAS and kidney --- p.58 / Chapter 3.1.6 --- Summary --- p.58 / Chapter 3.2 --- Materials and methods / Chapter 3.2.1 --- Experimental animals --- p.61 / Chapter 3.2.2 --- Salinity adaptation --- p.61 / Chapter 3.2.3 --- Drinking rate measurement --- p.61 / Chapter 3.2.4 --- Determination of angiotensin converting enzyme (ACE) activity --- p.61 / Chapter 3.2.5 --- Blood pressure experiment --- p.62 / Chapter 3.2.6 --- Statistical analysis --- p.63 / Chapter 3.3 --- Results / Chapter 3.3.1 --- Drinking rate --- p.64 / Chapter 3.3.2 --- ACE activity --- p.69 / Chapter 3.3.3 --- Blood pressure --- p.71 / Chapter 3.4 --- Discussion / Chapter 3.4.1 --- Drinking rate --- p.77 / Chapter 3.4.2 --- ACE activity --- p.81 / Chapter 3.4.3 --- Blood pressure --- p.83 / Chapter Chapter 4 --- General conclusion --- p.86 / Reference --- p.89

Zhang, Chaoxiong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 115-132). / Abstracts in English and Chinese. / Chapter I --- Title page --- p.i / Chapter II --- Thesis committee --- p.ii / Chapter III --- Abstract --- p.iii / Chapter IV --- Abstract (Chinese version) --- p.v / Chapter V --- Acknowledgement --- p.vii / Chapter VI --- Table of content --- p.viii / Chapter VII --- List of figure --- p.xiii / Chapter Chapter 1 --- General introduction --- p.1 / Chapter Chapter 2 --- Literature review --- p.7 / Chapter 2.1 --- An introduction to myostatin --- p.8 / Chapter 2.1.1 --- A general introduction --- p.8 / Chapter 2.1.2 --- Myostatin identification --- p.9 / Chapter 2.1.3 --- Structural studies of myostatin --- p.10 / Chapter 2.1.4 --- Phenotype of myostatin-null animals or transgenic animal --- p.10 / Chapter 2.2 --- Regulation of myostatin --- p.12 / Chapter 2.2.1 --- Biosynthesis of myostatin --- p.12 / Chapter 2.2.2 --- Regulation of myostatin expression --- p.13 / Chapter 2.2.3 --- Regulation of myostatin protein --- p.16 / Chapter 2.3 --- Myostatin effect --- p.20 / Chapter 2.3.1 --- Myostatin Signaling Pathway --- p.20 / Chapter 2.3.2 --- Cellular Responses to Myostatin Signaling --- p.23 / Chapter 2.4 --- Possible functions in tissues other than muscle --- p.26 / Chapter 2.5 --- Myostatin in fishes --- p.27 / Chapter 2.5.1 --- Introduction of silver sea bream --- p.27 / Chapter 2.5.2 --- Studies carried out in fishes --- p.27 / Chapter 2.5.3 --- Possible novel functions of myostatin in fishes --- p.30 / Chapter Chapter 3 --- Characterization of myostatin gene in the silver seabream (Sparus sarba) --- p.31 / Chapter 3.1 --- Abstract --- p.32 / Chapter 3.2 --- Introduction --- p.33 / Chapter 3.3 --- Materials and methods --- p.35 / Chapter 3.3.1 --- Experimental fish --- p.35 / Chapter 3.3.2 --- Total RNA extraction and cDNA cloning of myostatin-1 and myostatin-2 in silver sea bream --- p.35 / Chapter 3.3.3 --- Multiple sequence alignment --- p.38 / Chapter 3.3.4 --- Real-time PCR for quantification of myostatin-1 and myostatin-2 mRNA expression --- p.38 / Chapter 3.3.5 --- 1 --- p.39 / Chapter 3.3.6 --- Data processing and statistical analysis --- p.40 / Chapter 3.4 --- Results --- p.40 / Chapter 3.4.1 --- Cloning of myostatin-l and myostatin-2 cDNA --- p.40 / Chapter 3.4.2 --- Myostatin tissue distribution and seasonal pattern --- p.42 / Chapter 3.5 --- Discussion --- p.55 / Chapter Chapter 4 --- "Effects of growth hormone, 11-ketotestosterone and cortisol on myostatin mRNA expression in silver sea bream (Sparus sarba)" --- p.61 / Chapter 4.1 --- Abstract --- p.62 / Chapter 4.2 --- Introduction --- p.63 / Chapter 4.3 --- Materials and methods --- p.65 / Chapter 4.3.1 --- Experimental fish --- p.65 / Chapter 4.3.2 --- Growth hormone injection --- p.65 / Chapter 4.3.3 --- 11-ketotestosterone and cortisol injection --- p.66 / Chapter 4.3.4 --- Muscle explants culture and hormone exposure --- p.67 / Chapter 4.3.5 --- Primary pituitary cell culture and cortisol exposure --- p.68 / Chapter 4.3.6 --- Measurement of growth hormone secretion by ELISA --- p.69 / Chapter 4.3.7 --- Data processing and statistical analysis --- p.70 / Chapter 4.4 --- Results --- p.71 / Chapter 4.4.1 --- Growth hormone injection --- p.71 / Chapter 4.4.2 --- 11-ketotestosterone injection --- p.71 / Chapter 4.4.3 --- Cortisol injection --- p.71 / Chapter 4.4.4 --- "In vitro hormone treatment-growth hormone, 11-ketotestosterone and cortisol" --- p.72 / Chapter 4.4.5 --- Pituitary cell growth hormone secretion under cortisol treatment --- p.72 / Chapter 4.5 --- Discussion --- p.81 / Chapter Chapter 5 --- Expression of myostatin mRNA in silver sea bream in different salinity --- p.87 / Chapter 5.1 --- Abstract --- p.88 / Chapter 5.2 --- Introduction --- p.89 / Chapter 5.3 --- Materials and Methods --- p.91 / Chapter 5.3.1 --- Experimental fish --- p.92 / Chapter 5.3.2 --- Long term salinity adaptation --- p.92 / Chapter 5.3.3 --- Abrupt transfer form seawater to freshwater --- p.92 / Chapter 5.3.4 --- Data processing and statistical analysis --- p.93 / Chapter 5.4 --- Results --- p.93 / Chapter 5.4.1 --- Long term adaptation to different salinities --- p.93 / Chapter 5.4.2 --- Abrupt transfer from 33ppt to 6ppt - 24 h --- p.93 / Chapter 5.4.3 --- Abrupt transfer from 33ppt to 6ppt - 72 h --- p.94 / Chapter 5.5 --- Discussion --- p.104 / Chapter Chapter 6 --- General discussion and conclusion --- p.108 / References --- p.115

Yuen, Wing Sum. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 136-155). / Abstract also in Chinese. / Chapter I --- Title page --- p.i / Chapter II --- Acknowledgements --- p.ii / Chapter III --- Abstract --- p.iii / Chapter IV --- Abstract (Chinese version) --- p.vi / Chapter V --- Table of contents --- p.viii / Chapter VI --- List of abbreviations --- p.xv / Chapter VII --- List of figures --- p.xvi / Chapter Chapter 1 --- General introduction --- p.1 / Chapter Chapter 2 --- Literature review --- p.5 / Chapter 2.1 --- Cystic fibrosis transmembrane conductance regulator in human --- p.5 / Chapter 2.1.1. --- Pathology of cystic fibrosis --- p.5 / Chapter 2.1.2. --- CFTR gene and the encoded protein --- p.6 / Chapter 2.1.3. --- Hypothetical model for CFTR function --- p.7 / Chapter 2.1.4. --- Functions of CFTR --- p.7 / Chapter 2.1.5. --- Regulation of CFTR gene expression --- p.8 / Chapter 2.1.6 --- Regulation of CFTR protein --- p.9 / Chapter 2.1.7. --- Discovery of CFTR homologues in other vertebrates --- p.10 / Chapter 2.2 --- Cystic fibrosis transmembrane conductance regulator in teleosts --- p.10 / Chapter 2.2.1. --- Evidence for the presence of CFTR homologue in teleosts --- p.10 / Chapter 2.2.2. --- Molecular cloning of teleost CFTR genes --- p.11 / Chapter 2.2.3. --- Role of teleost CFTR in osmoregulation --- p.13 / Chapter 2.2.3.1. --- Tissue distribution of CFTR --- p.13 / Chapter 2.2.3.2. --- Changes in CFTR expression in response to ambient salinities --- p.14 / Chapter 2.2.3.3. --- Immunocytochemical studies of CFTR --- p.15 / Chapter 2.2.3.4. --- Regulation of CFTR --- p.17 / Chapter 2.3 --- Osmoregulation in teleosts --- p.19 / Chapter 2.3.1. --- Importance of osmoregulation --- p.19 / Chapter 2.3.2. --- Major components of chloride cells in marine teleosts --- p.20 / Chapter 2.3.2.1. --- Overview --- p.20 / Chapter 2.3.2.2. --- Sodium-potassium adenosine triphosphatase (Na+，K+-ATPase) --- p.21 / Chapter 2.3.2.3. --- Cystic fibrosis transmembrane conductance regulator (CFTR) --- p.22 / Chapter 2.3.2.4. --- Na+/K+/2Cr cotransporter (NKCC) --- p.23 / Chapter 2.3.2.5. --- Potassium (K+) channel --- p.25 / Chapter 2.4 --- Endocrine control of osmoregulation --- p.26 / Chapter 2.4.1. --- Overview --- p.26 / Chapter 2.4.2. --- Growth hormone (GH) and insulin-like growth factor I (IGF-I) --- p.27 / Chapter 2.4.2.1. --- Role of GH in osmoregulation --- p.27 / Chapter 2.4.2.2. --- Mediation through IGF-I --- p.29 / Chapter 2.4.2.3. --- Synergic effect with cortisol --- p.30 / Chapter 2.4.3. --- Prolactin (PRL) --- p.30 / Chapter 2.4.3.1. --- Role of PRL in osmoregulation --- p.30 / Chapter 2.4.3.2. --- Synergic effect with cortisol --- p.33 / Chapter 2.4.4. --- Cortisol --- p.33 / Chapter 2.4.4.1. --- Role of cortisol in osmoregulation --- p.33 / Chapter 2.4.4.2. --- Dual functions of cortisol --- p.34 / Chapter Chapter 3 --- Cloning and tissue distribution of silver sea bream CFTR gene --- p.36 / Chapter 3.1 --- Introduction --- p.36 / Chapter 3.2 --- Materials and methods --- p.38 / Chapter 3.2.1. --- Part A: Cloning of silver sea bream CFTR gene --- p.38 / Chapter 3.2.1.1. --- Fish and culture conditions --- p.38 / Chapter 3.2.1.2. --- Sampling of fish --- p.38 / Chapter 3.2.1.3. --- Preparation of first strand cDNA --- p.38 / Chapter 3.2.1.4. --- Design of primers --- p.39 / Chapter 3.2.1.5. --- Semi-quantitative reverse transcriptase (RT)-PCR --- p.40 / Chapter 3.2.1.6 --- Cloning --- p.41 / Chapter 3.2.2. --- Part B: Tissue distribution of CFTR in silver sea bream --- p.41 / Chapter 3.2.2.1. --- Fish and culture conditions --- p.41 / Chapter 3.2.2.2. --- Tissue sampling --- p.42 / Chapter 3.2.2.3. --- Preparation of first strand cDNA --- p.42 / Chapter 3.2.2.4 --- Design of primers --- p.42 / Chapter 3.2.2.5. --- Semi-quantitative reverse transcriptase (RT)-PCR --- p.43 / Chapter 3.3 --- Results --- p.44 / Chapter 3.3.1. --- Part A: Cloning of silver sea bream CFTR gene --- p.44 / Chapter 3.3.2. --- Part B: Tissue distribution of CFTR in silver sea bream --- p.60 / Chapter 3.4 --- Discussion --- p.62 / Chapter 3.4.1. --- Part A: Cloning of silver sea bream CFTR --- p.62 / Chapter 3.4.2. --- Part B: Tissue distribution of CFTR in silver sea bream --- p.64 / Chapter Chapter 4 --- Effect of salinity on CFTR mRNA expression in gill and posterior intestine of silver sea bream --- p.68 / Chapter 4.1 --- Introduction --- p.68 / Chapter 4.2 --- Materials and methods --- p.70 / Chapter 4.2.1. --- Part A: Effect of long-term exposure to different salinities on CFTR expression --- p.70 / Chapter 4.2.1.1. --- Experimental fish and salinity adaptation --- p.70 / Chapter 4.2.1.2. --- Tissue sampling --- p.70 / Chapter 4.2.1.3. --- Serum ion levels --- p.71 / Chapter 4.2.1.4. --- Preparation of first strand cDNA --- p.71 / Chapter 4.2.1.5. --- Design of primers --- p.71 / Chapter 4.2.1.6. --- Semi-quantitative reverse transcriptase (RT)-PCR --- p.71 / Chapter 4.2.1.7. --- Statistical analysis --- p.72 / Chapter 4.2.2. --- Part B: Effect of abrupt transfer on CFTR expression --- p.73 / Chapter 4.2.2.1. --- Experimental fish --- p.73 / Chapter 4.2.2.2. --- Experimental design --- p.73 / Chapter 4.2.2.2.1 --- Experiment 1: Abrupt transfer from seawater (SW) to 6 ppt --- p.73 / Chapter 4.2.2.2.2. --- Experiment 2: Abrupt transfer from 6 ppt to SW --- p.73 / Chapter 4.2.2.3. --- Tissue sampling --- p.74 / Chapter 4.2.2.4. --- Serum ion levels --- p.74 / Chapter 4.2.2.5. --- Preparation of first strand cDNA --- p.74 / Chapter 4.2.2.6. --- Design of primers --- p.75 / Chapter 4.2.2.7. --- Semi-quantitative reverse transcriptase (RT)-PCR --- p.75 / Chapter 4.2.2.8. --- Statistical analysis --- p.75 / Chapter 4.3 --- Results --- p.76 / Chapter 4.3.1. --- Part A: Effect of long-term exposure to different salinities on CFTR expression --- p.76 / Chapter 4.3.1.1. --- Serum ion levels --- p.76 / Chapter 4.3.1.2. --- CFTR expression in gill --- p.76 / Chapter 4.3.1.3. --- CFTR expression in posterior intestine --- p.76 / Chapter 4.3.2. --- Part B: Effect of abrupt salinity transfer on CFTR expression --- p.83 / Chapter 4.3.2.1. --- Experiment 1: Abrupt transfer from SW to 6 ppt --- p.83 / Chapter 4.3.2.1.1. --- Serum ion levels --- p.83 / Chapter 4.3.2.1.2. --- CFTR in gill --- p.83 / Chapter 4.3.2.1.3. --- CFTR in posterior intestine --- p.83 / Chapter 4.3.2.2. --- Experiment 2: Abrupt transfer from 6 ppt to SW --- p.89 / Chapter 4.3.2.2.1. --- Serum ion levels --- p.89 / Chapter 4.3.2.2.2. --- CFTR in gill --- p.89 / Chapter 4.3.2.2.3. --- CFTR in posterior intestine --- p.89 / Chapter 4.4 --- Discussion --- p.95 / Chapter 4.4.1. --- Long-term exposure to various salinities --- p.95 / Chapter 4.4.2. --- Abrupt salinity transfer --- p.98 / Chapter 4.4.2.1. --- Abrupt hypo-osmotic transfer (33 ppt to 6 ppt) --- p.98 / Chapter 4.4.2.2. --- Abrupt seawater transfer (6 ppt to 33 ppt) --- p.99 / Chapter 4.4.3. --- CFTR mRNA expression in posterior intestine --- p.101 / Chapter 4.4.4. --- Conclusion --- p.101 / Chapter Chapter 5 --- Effect of hormones on CFTR expression in gill and posterior intestine of silver sea bream --- p.102 / Chapter 5.1 --- Introduction --- p.102 / Chapter 5.2 --- Materials and methods --- p.104 / Chapter 5.2.1. --- Part A: In vivo effect of hormones on CFTR expression --- p.104 / Chapter 5.2.1.1. --- Experimental fish and salinity adaptation --- p.104 / Chapter 5.2.1.2. --- Hormone treatment --- p.104 / Chapter 5.2.1.3. --- Tissue sampling --- p.105 / Chapter 5.2.1.4. --- "Serum ion levels, preparation of first strand cDNA, design of primers and semi-quantitative reverse transcriptase (RT)-PCR" --- p.105 / Chapter 5.2.1.5. --- Statistical analysis --- p.105 / Chapter 5.2.2. --- Part B: In vitro effect of hormones on CFTR expression --- p.106 / Chapter 5.2.2.1. --- Fish and culture conditions --- p.106 / Chapter 5.2.2.2. --- Gill and posterior intestine preparations --- p.106 / Chapter 5.2.2.3. --- Hormone treatment --- p.106 / Chapter 5.2.2.4. --- "Preparation of first strand cDNA, design of primers and semi-quantitative reverse transcriptase (RT)-PCR" --- p.107 / Chapter 5.2.2.5. --- Statistical analysis --- p.107 / Chapter 5.3 --- Results --- p.108 / Chapter 5.3.1. --- Part A: In vivo effect of hormones on CFTR expression --- p.108 / Chapter 5.3.1.1. --- Serum ion levels --- p.108 / Chapter 5.3.1.1.1. --- Serum [Na+] level --- p.108 / Chapter 5.3.1.1.2. --- Serum [K+] level --- p.108 / Chapter 5.3.1.1.3. --- Serum [Cl' ] level --- p.108 / Chapter 5.3.1.2. --- CFTR expression in gill --- p.109 / Chapter 5.3.1.3. --- CFTR expression in posterior intestine --- p.109 / Chapter 5.3.2. --- Part B: In vitro effect of hormones on CFTR expression --- p.115 / Chapter 5.3.2.1. --- CFTR expression in gill --- p.115 / Chapter 5.3.2.2. --- CFTR expression in posterior intestine --- p.115 / Chapter 5.4 --- Discussion --- p.122 / Chapter 5.4.1. --- Effects of cortisol on CFTR expression --- p.122 / Chapter 5.4.2. --- Effects of growth hormone on CFTR expression --- p.124 / Chapter 5.4.3. --- Effects of prolactin on CFTR expression --- p.127 / Chapter 5.4.4. --- "Overall effect of cortisol, growth hormone and prolactin on CFTR expression" --- p.128 / Chapter 5.4.5 --- Conclusion --- p.130 / Chapter Chapter 6 --- General discussion and conclusion --- p.132 / References --- p.136

Chau, Kai Ming. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 136-159). / Abstract also in Chinese. / Chapter I --- Abstract --- p.i / Chapter II --- Acknowledgements --- p.vi / Chapter III --- Table of Contents --- p.vii / Chapter IV --- List of Figures --- p.xv / Chapter Chapter 1: --- Introduction --- p.1 / Chapter Chapter 2: --- Literature review --- p.7 / Chapter 2.1 --- Na+-K+ ATPase --- p.7 / Chapter 2.1.1 --- Introduction / Chapter 2.1.2 --- Structure of Na+-K+ ATPase --- p.9 / Chapter 2.1.1.2 --- Na+-K+ ATPase a subunit --- p.9 / Chapter 2.1.1.3 --- Na+-K+ ATPase β subunit --- p.11 / Chapter 2.1.1.4 --- Composition of the a subunit and β subunit --- p.12 / Chapter 2.1.1.5 --- Isomers of Na+-K+ ATPase --- p.13 / Chapter 2.1.1.6 --- Mechanism of ion exchange --- p.15 / Chapter 2.2 --- Aquaporins --- p.17 / Chapter 2.2.1 --- Introduction --- p.17 / Chapter 2.2.2 --- Structure of AQP-1 --- p.18 / Chapter 2.2.3 --- Distribution and function of AQP-1 --- p.19 / Chapter 2.3 --- Hormone --- p.22 / Chapter 2.3.1 --- Prolactin --- p.22 / Chapter 2.3.1.1 --- Structure of prolactin --- p.22 / Chapter 2.3.1.2. --- Functions of prolactin --- p.24 / Chapter 2.3.2 --- Growth hormone --- p.27 / Chapter 2.3.2.1 --- Structure --- p.27 / Chapter 2.3.2.2 --- Function of growth hormone --- p.28 / Chapter 2.3.3 --- Cortisol --- p.30 / Chapter 2.3.3.1 --- Structure --- p.30 / Chapter 2.3.3.2 --- Functions of cortisol --- p.31 / Chapter 2.4 --- Sparus sarba --- p.34 / Chapter 2.5 --- Urinary bladder of fish --- p.36 / Chapter Chapter 3: --- Effect of salinity on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder of silver sea bream Sparus sarba --- p.38 / Chapter 3.1 --- Introduction --- p.38 / Chapter 3.2 --- Chronic effect of salinity on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.40 / Chapter 3.2.1 --- Materials and Methods --- p.40 / Chapter 3.2.1.1 --- Fish --- p.40 / Chapter 3.2.1.2 --- Tissue sampling --- p.41 / Chapter 3.2.1.3 --- Protein extraction and quantification --- p.41 / Chapter 3.2.1.4 --- Na+-K+ ATPase ATPase activity --- p.42 / Chapter 3.2.1.5 --- RNA extraction and first strand cDNA synthesis --- p.43 / Chapter 3.2.1.6 --- Validation of semi-quantitative RT-PCR --- p.45 / Chapter 3.2.1.7 --- Semi-quantification of expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.47 / Chapter 3.2.1.8 --- Statistical analysis --- p.47 / Chapter 3.2.2 --- Results --- p.48 / Chapter 3.2.2.1 --- Na+-K+ ATPase activity --- p.48 / Chapter 3.2.2.2 --- Relative expression of Na+-K+ ATPase and aquaporin-1 in urinary bladder --- p.48 / Chapter 3.2.3 --- Discussion --- p.54 / Chapter 3.2.3.1 --- Chronic effect of salinity on Na+-K+ ATPase in urinary bladder --- p.54 / Chapter 3.2.3.2 --- Chronic effect of salinity on AQP-1 expression in urinary bladder --- p.59 / Chapter 3.3 --- Effect of abrupt transfer on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.61 / Chapter 3.3.1. --- Materials and Methods --- p.61 / Chapter 3.3.1.1 --- Fish --- p.61 / Chapter 3.3.1.2 --- Tissue sampling --- p.62 / Chapter 3.3.1.3 --- "RNA extraction, first strand cDNA synthesis and RT-PCR" --- p.62 / Chapter 3.3.1.4 --- Statistical analysis --- p.63 / Chapter 3.3.2 --- Results --- p.64 / Chapter 3.3.2.1 --- Effect of abrupt hypo-osmotic transfer on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.64 / Chapter 3.3.2.2 --- Effect of abrupt hyper-osmotic transfer on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.65 / Chapter 3.3.3 --- Discussion --- p.73 / Chapter 3.4 --- Effect of in vitro salinity on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.78 / Chapter 3.4.1 --- Materials and Methods --- p.78 / Chapter 3.4.1.1 --- Fish --- p.78 / Chapter 3.4.1.2 --- Tissue sampling --- p.78 / Chapter 3.4.1.3 --- Preparation of culture medium --- p.79 / Chapter 3.4.1.4 --- "RNA extraction, first strand cDNA synthesis and RT-PCR" --- p.79 / Chapter 3.4.1.5 --- Statistical analysis --- p.80 / Chapter 3.4.2 --- Results --- p.81 / Chapter 3.4.3 --- Discussion --- p.85 / Chapter 3.5 --- Conclusion --- p.86 / Chapter Chapter 4: --- Effect of hormones on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder of silver sea bream Sparus sarba --- p.88 / Chapter 4.1 --- Introduction --- p.88 / Chapter 4.2 --- In vivo effect of hormones on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder of silver sea bream Sparus sarba --- p.91 / Chapter 4.2.1 --- Material and method --- p.91 / Chapter 4.2.1.1 --- Fish --- p.91 / Chapter 4.2.1.2 --- Tissue sampling --- p.92 / Chapter 4.2.1.3 --- "RNA extraction, first strand cDNA synthesis and RT-PCR" --- p.92 / Chapter 4.2.1.4 --- Statistical analysis --- p.92 / Chapter 4.2.2 --- Results / Chapter 4.2.2.1 --- Hormonal effect on mRNA expression of Na+-K+ ATPase and AQP-1 in urinary bladder of sea water adapted fish --- p.93 / Chapter 4.2.2.2 --- Hormonal effect on mRNA expression of Na+-K+ ATPase and AQP-1 in urinary bladder of brackish water adapted fish --- p.97 / Chapter 4.2.3 --- Discussion --- p.101 / Chapter 4.2.3.1 --- Effect of prolactin on mRNA expression of Na+-K+ ATPase and AQP-1 in urinary bladder --- p.101 / Chapter 4.2.3.2 --- Effect of growth hormone on mRNA expression of Na+-K+ ATPase and AQP-1 in urinary bladder --- p.104 / Chapter 4.2.3.3 --- Effect of cortisol on mRNA expression of Na+-K+ ATPase and AQP-1 in urinary bladder --- p.106 / Chapter 4.3 --- In vitro effect of hormone on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder of silver sea bream Sparus sarba --- p.109 / Chapter 4.3.1 --- Materials and methods --- p.109 / Chapter 4.3.1.1 --- Fish --- p.109 / Chapter 4.3.1.2 --- Tissue sampling --- p.110 / Chapter 4.3.1.3 --- Preparation of culture medium --- p.110 / Chapter 4.3.1.4 --- "RNA extraction, first strand cDNA synthesis and RT-PCR" --- p.111 / Chapter 4.3.1.5 --- Statistical analysis --- p.111 / Chapter 4.3.2 --- Results --- p.112 / Chapter 4.3.2.1 --- Effect of prolactin on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.112 / Chapter 4.3.2.2 --- Effect of growth hormone on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.113 / Chapter 4.3.2.3 --- Effect of cortisol on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.123 / Chapter 4.3.3 --- Discussion --- p.124 / Chapter 4.3.3.1 --- Effect of prolactin on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.124 / Chapter 4.3.3.2 --- Effect of growth hormone on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.125 / Chapter 4.3.3.3 --- Effect of cortisol on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.127 / Chapter 4.4 --- Conclusion --- p.129 / Chapter Chapter 5 --- General Conclusions --- p.131 / References --- p.136