Zhu Jinxia. / "July 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 142-170). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

In a HEK 293 cell line stably expressing seabream GHS-R1a (sbGHS-R1a), we found that a synthetic growth hormone secretagogue (GHS) increased [ 3H]-inositol phosphate production, clearly indicating coupling of this receptor to Gq/11-proteins. Using Western blotting, we found that GHS could also stimulate extracellular signal-regulated kinases 1 and 2 (ERK1/2), and that this response was inhibited by the MEK inhibitor U0126. For both the [3H]-inositol phosphate and ERK1/2 assays, the presence of the GHS-R antagonist D-Lys(3)-GHRP-6 significantly inhibited the GHS-stimulated activities, and in addition inhibited basal activities by 50% and 40%, respectively. These results showed that sbGHS-R1a is a constitutively active receptor and the antagonist D-Lys(3)-GHRP-6 is an inverse agonist. We also proposed that the expression of sbGHS-Rs was involved in the regulation of cell apoptosis. / Oligomerization of the human GHS-Rs (hGHS-Rs) was explored by transient transfection of the hGHS-Rs in HEK 293 cells followed by co-immunoprecipitation of differentially epitope-tagged forms of the receptors and bioluminescence resonance energy transfer 2 (BRET2) studies. (Abstract shortened by UMI.) / The concept that G protein-coupled receptors (GPCRs) exist and potentially function as dimers and/or higher oligomers has progressed from hypothesis to being widely accepted recently. Oligomerization of GPCRs has been increasingly noted in the regulation of the biological activity of the receptors. The growth hormone secretagogue receptor 1a (GHS-R1a) is a GPCR which principally regulates the pulsatile release of growth hormone from the pituitary gland. The GHS-R exists in two forms: GHS-R1a being a constitutively-active GPCR with 7 transmembrane (TM) domains, and GHS-R1b being a truncated version of type 1a but having only 5 TM domains. The endogenous agonist for GHS-R1a is ghrelin which exerts a wide range of physiological actions, but the function of GHS-R1b is still unclear. Since the tissue distribution patterns of the two isoforms of GHS-R are different, the objective of the present study is to explore the mechanisms of cell signalling of GHS-R1a and to determine the extent and importance of interactions between these two receptor isoforms. / Leung Po Ki. / "July 2005." / Adviser: Helen Wise. / Source: Dissertation Abstracts International, Volume: 67-07, Section: B, page: 3728. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (p. 189-210). / 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. / School code: 1307.

Kwan Hiu Yee. / "June 2006." / 2+ in the title is superscript. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 131-150). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese.

Wong, Chun Kit. / "September 2010." / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 104-118). / Abstracts in English and Chinese. / Thesis Committee --- p.i / Acknowledgements --- p.ii / Contents --- p.iii / Declaration --- p.vi / Abstract --- p.vii / 摘要 --- p.x / Abbreviations --- p.xi / List of Figures --- p.xiii / Chapter CHAPTER ONE: --- INTRODUCTION / Chapter 1.1 --- Embryonic Stem Cells (ESCs) / Chapter 1.1.1 --- Characteristics of ESC --- p.1 / Chapter 1.1.2 --- Therapeuticotential of ESCs --- p.2 / Chapter 1.2 --- 17β-estradiol (E2) / Chapter 1.2.1 --- Genomic Actions of E2 --- p.3 / Chapter 1.2.2 --- Non-genomic Actions of E2 --- p.5 / Chapter 1.2.3 --- hysiological Roles of E2 on Early Mammalian Development --- p.9 / Chapter 1.2.4 --- E2 and Cell Proliferation --- p.10 / Chapter 1.3 --- Ca2+ homeostasis / Chapter 1.3.1 --- Overview --- p.11 / Chapter 1.3.2 --- Ca2+ Signaling in mESCs --- p.14 / Chapter 1.4 --- Store-operated Ca2+ Entry (SOCE) / Chapter 1.4.1 --- Overview --- p.15 / Chapter 1.4.2 --- Store Depletion --- p.15 / Chapter 1.4.3 --- Activation of SOCE --- p.16 / Chapter 1.5 --- Molecular Identities of Store-operated Ca2+ Channels (SOCCs) on plasma Membrane / Chapter 1.5.1 --- TRPC Channels --- p.17 / Chapter 1.5.2 --- ORAI Channels --- p.18 / Chapter 1.5.3 --- Regulation of SOCCs at Different Levels --- p.18 / Chapter 1.5.4 --- Regulation of SOCE --- p.19 / Chapter 1.6 --- Nuclear Factor of Activated T-cells (NFAT) / Chapter 1.6.1 --- Overview --- p.20 / Chapter 1.6.2 --- Mechanisms of Action --- p.21 / Chapter 1.6.3 --- Functions --- p.22 / Chapter 1.7 --- Aims of the Study --- p.23 / Chapter CHAPTER TWO: --- MATERIALS AND METHODS / Chapter 2.1 --- Maintenance of mESCs --- p.24 / Chapter 2.2 --- Cell proliferation Assay and Viability Test --- p.24 / Chapter 2.3 --- "RNAreparation, Reverse Transcription (RT) and Quantitative Polymerase Chain Reaction (qPCR)" --- p.25 / Chapter 2.4 --- Totalrotein Extraction --- p.27 / Chapter 2.5 --- Measurement of protein Concentration --- p.27 / Chapter 2.6 --- De-phosphorylation Assay --- p.28 / Chapter 2.7 --- Western Blot --- p.28 / Chapter 2.8 --- Ca2+ Measurement by Confocal Microscopy --- p.30 / Chapter 2.9 --- Ca2+ Measurement by Flow Cytometry --- p.31 / Chapter 2.10 --- siRNA Transfection --- p.31 / Chapter 2.11 --- DNAlasmid Transfection --- p.32 / Chapter 2.12 --- Molecular and Fluorescence Imaging --- p.33 / Chapter 2.13 --- Statistical Analysis --- p.34 / Chapter 2.14 --- Primers used in the Study (Table 1:Primers List) --- p.34 / Chapter 2.15 --- Drugs used in the Study (Table 2: Drugs List) --- p.36 / Chapter 2.16 --- Antibodies used in the Study (Table 3: Antibodies List) --- p.37 / Chapter CHAPTER THREE: --- RESULTS / Chapter 3.1 --- Expression of SOCE in mESCs --- p.38 / Chapter 3.2 --- SOCC Blockers Attenuated mESCroliferation --- p.43 / Chapter 3.3 --- E2 Increased mESCroliferation --- p.48 / Chapter 3.4 --- E2 Increased Intracellular Ca2+ ([Ca2+]i) Level in mESCs --- p.48 / Chapter 3.5 --- E2 Increased the Amplitude of SOCE --- p.51 / Chapter 3.6 --- Increase in mESC proliferation and SOCE Caused by E2 Could be Reversed by SOCC Blocker --- p.51 / Chapter 3.7 --- Relative Expression of SOCC Candidates at mRNA Level Under the Treatment of E2 --- p.56 / Chapter 3.8 --- E2 Down-regulated the Expression of ORAI3 --- p.56 / Chapter 3.9 --- Knockdown of ORAI3 in mESCs --- p.61 / Chapter 3.10 --- Identification of NFATc3 Specific Bands --- p.63 / Chapter 3.11 --- E2 Increased the phosphorylation of NFATc3 --- p.67 / Chapter 3.12 --- Effects of 2-APB on NFATc3 phosphorylation Status --- p.67 / Chapter 3.13 --- Identification of NFATc4 Specific Bands ？ --- p.72 / Chapter 3.14 --- E2 Increased the Translocation of GFP-NFATc4 From the Cytoplasm to the Nucleus and This Effect Could be Reversed by 2-APB --- p.80 / Chapter 3.15 --- CsA Reversed E2-induced Increase in proliferation --- p.82 / Chapter CHAPTER FOUR: --- DISCUSSION / Chapter 4.1 --- Expression of SOCE in mESCs --- p.84 / Chapter 4.2 --- proliferation of mESCs Depends on SOCE --- p.85 / Chapter 4.3 --- E2 Acts an Extrinsic Factor for Stimulatingroliferation of mESCs Via SOCE --- p.87 / Chapter 4.4 --- roposed Mechanism to Show an Increment of SOCE Can be Due to a Down-regulation of ORAI3 --- p.89 / Chapter 4.5 --- Experiments Aiming to Knockdown ORAI3 --- p.92 / Chapter 4.6 --- roposed Mechanism to Show an Increment of SOCE by Other SOCC Candidates Rather than ORAI3 --- p.93 / Chapter 4.7 --- Activation of NFATc3 and NFATc4 by E2 in mESCs --- p.94 / Chapter 4.8 --- possible Downstream Targets of NFAT Responsible for E2-induced mESCs proliferation --- p.96 / Chapter CHAPTER FIVE: --- FUTUREERSPECTIVES --- p.98 / Chapter CHAPTER SIX: --- CONCLUSION --- p.100 / REFERENCES --- p.104

Thesis (Ph. D.)--University of Missouri--Columbia, 2002. / Typescript. Includes bibliographical references. Also available on the Internet.

Thesis (Ph.D.)--Memorial University of Newfoundland, Faculty of Medicine, 2001. / Typescript. Bibliography: leaves 188-241.

Thesis (M.S.)--West Virginia University, 2000. / Title from document title page. Document formatted into pages; contains ix, 70 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references (p. 63-68).

Neuronal connections in cat visual cortex are highly susceptible to visual experience at early postnatal age and thus serve as a useful model of neural plasticity. The biochemical mechanisms underlying this cortical plasticity remain unclear. In this thesis, the development of several elements in calcium-related signal transduction systems, including the type-1 muscarinic and alpha-1 adrenoceptor systems as examples of cell surface receptors and protein kinase C. calcium/calmodulin dependent kinase II and inositol 1,4,5 phosphotate receptors as second messenger targets, were investigated using the methods of immunocytochemistry and autoradiography. The results show that each receptor develops with its own time-table and laminar distribution; the various elements all culminate and display the maximal colocalization during the critical period; and, only at this age, the cortical levels of the receptors and kinases are dependent on subcortical afferents. The results suggest that cell surface receptors and their second messenger targets develop in specific temporal and spatial patterns, which may be both genetically and environmentally determined, and this specific sequence of development of the molecules for signal transduction results in a series of modifications in the morphology and physiology of the developing cortex leading to its maturation. / Medicine, Faculty of / Graduate

<p dir="ltr">Phospholipase Cε (PLCε) is required for normal cardiovascular function, and dysregulation of its expression or activity has been shown to cause cardiac hypertrophy and heart failure. However, regulation of PLCε by the RhoA small GTPase protects the heart against ischemia-reperfusion injury, particularly downstream of G_12/13-coupled receptors. Despite the role of RhoA and PLCε in driving the cardioprotective response, little is known about how these proteins interact to increase lipase activity.<b> </b>RhoA was initially thought to bind to PLCε through one of its C-terminal Ras association (RA) domains, which are essential for its regulation by other GTPases. However, the RA domains are dispensable for both RhoA binding and activation, and further truncations of PLCε narrowed its binding site to the highly conserved PLC catalytic core. Functional studies implicated an insertion within the catalytic TIM barrel domain, known as the Y-box, as a requirement for RhoA-dependent activation of PLCε. However, the Y-box does not bind the GTPase. The goal of this dissertation is to identify the molecular mechanism by which RhoA binds to PLCε and increases its activity using structural and functional studies. The successful completion of these studies will map the interaction between these two critical signaling proteins, as well as identify elements in PLCε required for activation at the membrane. Ultimately, this knowledge can be exploited to develop lead therapeutic compounds that modulate this interaction to improve cardiovascular health.</p>

Tang, Yongjiang. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 159-202). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.