published_or_final_version / Microbiology / Master / Master of Medical Sciences

by Liu Heng. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 115-138). / Acknowledgments --- p.I / List of Abbreviations --- p.II / Abstract --- p.V / Chapter Chapter 1: --- Introduction / Chapter 1.1 --- Cytokines in the Central Nervous System --- p.1 / Chapter 1.1.1 --- Basic Properties of Cytokines --- p.1 / Chapter 1.1.2 --- The General Characteristics of Glial Cells --- p.4 / Chapter 1.1.2.1 --- Astrocytes --- p.4 / Chapter 1.1.2.2 --- Oligodendrocytes --- p.6 / Chapter 1.1.2.3 --- Microglial --- p.7 / Chapter 1.1.3 --- The Effects of Cytokines on Neural Cells --- p.7 / Chapter 1.1.3.1 --- TNF-α and Neural Cells --- p.8 / Chapter 1.1.3.2 --- LIF and Neural Cells --- p.10 / Chapter 1.1.3.3 --- IL-1 and Neural Cells --- p.12 / Chapter 1.1.3.4 --- IL-6 and Neural Cells --- p.14 / Chapter 1.1.4 --- Immune Response in the Central Nervous System --- p.16 / Chapter 1.2 --- The C6 Glioma as a Model for the Study of Glial Cell Growth and Differentiation --- p.21 / Chapter 1.2.1 --- The Rat C6 Glioma Cells --- p.21 / Chapter 1.2.2 --- The Differentiation and Proliferation of C6 Glioma Cells --- p.23 / Chapter 1.3 --- Signal Transduction Pathways in Cytokine-stimulated Glial Cells --- p.28 / Chapter 1.3.1 --- Intracellular Signalling Pathways of Cytokines --- p.28 / Chapter 1.3.1.1 --- Protein Kinase C Pathway --- p.29 / Chapter 1.3.1.2 --- Tyrosine Kinase Pathway --- p.30 / Chapter 1.3.1.3 --- Cyclic Nucleotide Pathway --- p.32 / Chapter 1.3.1.4 --- Nitric Oxide Pathway --- p.33 / Chapter 1.3.2 --- Intracellular Signalling Pathways in Cytokine-stimulated C6 Glioma Cells --- p.34 / Chapter 1.4 --- The Aims of This Thesis Project --- p.37 / Chapter Chapter 2: --- Materials and Methods --- p.41 / Chapter 2.1 --- Rat C6 Glioma Cell Culture --- p.41 / Chapter 2.1.1 --- Preparation of Culture Media --- p.41 / Chapter 2.1.1.1 --- Complete Dulbecco's Modified Eagle Medium --- p.41 / Chapter 2.1.1.2 --- Complete Roswell Park Memorial Institute1640 Medium --- p.42 / Chapter 2.1.2 --- Maintenance of the C6 Cell Line --- p.42 / Chapter 2.1.3 --- Cell Preparation for Assays --- p.43 / Chapter 2.2 --- Determination of Cell Proliferation --- p.44 / Chapter 2.2.1 --- Determination of Cell Proliferation by [3H]-Thymidine Incorporation --- p.44 / Chapter 2.2.2 --- Measurement of Cell Viability Using Neutral Red Assay --- p.45 / Chapter 2.2.3 --- Data Analysis --- p.45 / Chapter 2.3 --- Effects of Cytokines and Lipopolysaccharide on C6 Cell Proliferation --- p.46 / Chapter 2.4 --- Effects of Protein Kinase C Activators and Inhibitors on Cytokine-induced C6 Cell Proliferation --- p.47 / Chapter 2.5 --- Effects of cAMP or cGMP on Cytokine-induced C6 Cell Proliferation --- p.48 / Chapter 2.6 --- Effects of Tyrosine Kinase Inhibitors on Cytokine-induced C6 Cell Proliferation --- p.48 / Chapter 2.7 --- Effects of Calcium Ion on Cytokine-induced C6 Cell Proliferation --- p.49 / Chapter 2.8 --- Effects of Nitric Oxide on Cytokine-induced C6 Cell Proliferation --- p.49 / Chapter 2.8.1 --- Effects of Sodium Nitroprusside and Nitric Oxide Synthase Inhibitors on Cytokine-induced C6 Cell Proliferation --- p.49 / Chapter 2.8.2 --- Nitric Oxide Production Assay --- p.50 / Chapter 2.9 --- Effects of β-Adrenergic Receptor Agonist and Antagonist on Cytokine-induced C6 Cell Proliferation --- p.51 / Chapter 2.10 --- Morphological Studies on Cytokine-Treated C6 Glioma Cells --- p.51 / Chapter 2.10.1 --- Wright-Giesma Staining --- p.52 / Chapter 2.10.2 --- Glial Fibrillary Acidic Protein Staining --- p.52 / Chapter 2.10.3 --- Hematoxylin Staining --- p.53 / Chapter Chapter 3: --- Results --- p.55 / Chapter 3.1 --- Effects of Cytokines on C6 Cell Proliferation --- p.55 / Chapter 3.1.1 --- Effects of Cytokines on C6 Cell Proliferation --- p.56 / Chapter 3.1.2 --- The Time Course of Cytokine-induced C6 Cell Proliferation --- p.59 / Chapter 3.1.3 --- Effects of Lipopolysaccharide on C6 Cell Proliferation --- p.61 / Chapter 3.1.4 --- Effects of Cytokines on the Growth of C6 Cells --- p.64 / Chapter 3.2 --- Morphology and GFAP Expression in Cytokine-treated C6 Glioma Cells --- p.64 / Chapter 3.2.1 --- Effects of Cytokines on the Morphology of C6 Cells --- p.64 / Chapter 3.2.2 --- Effects of Cytokines on GFAP Expression in C6 Glioma Cells --- p.66 / Chapter 3.3 --- The Signalling Pathway of Cytokine-induced C6 Cell Proliferation --- p.69 / Chapter 3.3.1 --- The Involvement of Protein Kinase C in Cytokine-induced C6Cell Proliferation --- p.71 / Chapter 3.3.2 --- The Involvement of Tyrosine Kinase in the Cytokine- induced C6 Cell Proliferation --- p.81 / Chapter 3.3.3 --- The Involvement of Calcium Ions in Cytokine-induced C6 Cell Proliferation --- p.87 / Chapter 3.3.4 --- The Involvement of Cyclic Nucleotides in Cytokine- induced C6 Cell Proliferation --- p.92 / Chapter 3.3.5 --- The Involvement of Nitric Oxide in Cytokine-induced C6 Cell proliferation --- p.94 / Chapter 3.3.6 --- The Involvement of P-Adrenergic Receptor in Cytokine- induced C6 Cell Proliferation --- p.101 / Chapter Chapter 4: --- Discussion and Conclusions --- p.104 / References --- p.115

The suppressor of cytokine signaling ( SOCS ) proteins have been identified as important regulators of cytokine signaling. SOCS - 3 has been identified as being essential for normal fetal growth and survival, with the null mutation of the socs - 3 gene resulting in embryo death. The specific role of SOCS - 3 in fetal development, however, has yet to be characterized. Therefore, the overall aim of this thesis was to identify and quantify SOCS - 3 mRNA in a range of fetal tissues in the sheep. After identification of SOCS - 3 expression in fetal tissues, we then aimed to determine the ontogenic profile of SOCS - 3 in three key fetal tissues ; the liver, adipose tissue and adrenal gland, and whether SOCS - 3 expression in these tissues was altered after withdrawal and stimulation of prolactin ( PRL ). SOCS - 3 mRNA was found to be differentially expressed in a range of fetal tissues in late gestation and was higher in the fetal liver than in the pancreas, spleen and kidney. SOCS - 3 expression increased throughout gestation in the fetal liver, however, its expression decreased in the fetal adipose tissue and adrenal in late gestation. The pituitary hormone PRL has previously been implicated as a fetal growth factor. In the sheep fetus, PRL receptors are expressed in the fetal liver, adipose tissue and adrenal. We aimed to determine whether PRL plays a role in the maintenance of SOCS - 3 expression in the liver, adipose tissue and adrenal gland in late gestation, and whether SOCS - 3 expression can be regulated by acute PRL stimulation. We have demonstrated that PRL withdrawal suppressed SOCS - 3 expression in the liver, whereas acute PRL stimulation upregulated SOCS - 3 expression in the adrenal. Neither PRL withdrawal nor stimulation had an effect on SOCS - 3 expression in the adipose tissue. In summary, the data presented in this thesis would suggest that SOCS - 3 has tissue specific functions in late gestation. Furthermore, its expression is regulated in a tissue specific manner in response to the withdrawal or acute stimulation by PRL This provides the first evidence to suggest that the fetal liver and adrenal are both sensitive to either chronic or acute changes in plasma PRL concentrations, measured as the suppression or upregulation of SOCS - 3. We speculate that changes in SOCS - 3 mRNA expression relates to the regulation of growth and functional maturation of fetal tissues throughout gestation, and that PRL may represent an important factor which acts to alter SOCS - 3 expression in key fetal tissues. / Thesis (Ph.D.)--School of Molecular and Biomedical Science, 2006.

Bibliography: leaves 151-179. / xx, 179 leaves : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Examines aspects of the distribution of leukocyte subpopulation in human corpus luteum, cytokine determination in human preovulatory follicular fluid, as well as the effects of cytokines on human granulosalutein cells; with the aim of investigating one of the ovarian regulatory systems, which may be controlled by immune cell-derived cytokines. / Thesis (Ph.D.)--University of Adelaide, Dept. of Obstetrics and Gynaecology, 1993

published_or_final_version / Biological Sciences / Doctoral / Doctor of Philosophy

Addendum pasted on front end papers. / Includes bibliographies. / 109, [99] leaves, [5] leaves of plates : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Describes the investigation as to whether the mouse granulocyte-macrophage colony-stimulating factor and interleukin-3 genes are regulated in a similar manner as those of the human, focussing on regulation through an enhancer. / Thesis (Ph.D.)--University of Adelaide, Dept. of Microbiology and Immunology, 1996