Ip Lai Fong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 81-92). / Abstracts in English and Chinese. / Table of Contents --- p.1 / Abbreviations --- p.4 / Chapter Chapter 1 --- Introduction and Background --- p.5 / Chapter 1.1 --- Endothelin system --- p.5 / Chapter 1.1.1 --- Endothelins --- p.5 / Chapter 1.1.2 --- Endothelin converting enzyme (ECE) isoforms --- p.12 / Chapter 1.1.3 --- Endothelin receptors --- p.24 / Chapter 1.2 --- Signal-transduction mechanisms in ET system --- p.27 / Chapter 1.3 --- The aim of the present thesis --- p.31 / Chapter Chapter 2 --- Materials and Methods --- p.32 / Chapter 2.1 --- Primer Design --- p.32 / Chapter 2.2 --- Total RNA Isolation --- p.33 / Chapter 2.3 --- Reverse transcriptase polymerase chain reaction (RT-PCR) --- p.34 / Chapter 2.3.1 --- First Strand cDNA Synthesis --- p.34 / Chapter 2.3.2 --- PCR reaction --- p.34 / Chapter 2.4 --- Agarose gel electrophoresis --- p.35 / Chapter 2.5 --- Ligation of PCR inserts to cloning vector by TA cloning method --- p.35 / Chapter 2.6 --- Competent cell preparation --- p.36 / Chapter 2.7 --- Transformation and Screening --- p.37 / Chapter 2.8 --- Plasmid DNA Extraction --- p.38 / Chapter 2.9 --- DNA sequencing --- p.38 / Chapter 2.10 --- DIG RNA Labeling --- p.38 / Chapter 2.10.1 --- Plasmid Linearization --- p.38 / Chapter 2.10.2 --- Transcription --- p.39 / Chapter 2.10.3 --- Probe purification --- p.39 / Chapter 2.11 --- In situ hybridizaion --- p.40 / Chapter 2.11.1 --- Tissue preparation and slide mounting --- p.40 / Chapter 2.11.2 --- Non-radioactive in situ hybridization --- p.41 / Chapter 2.12 --- Whole Mount non-radioactive in situ hybridization --- p.42 / Chapter 2.12.1 --- Dissection and fixation --- p.42 / Chapter 2.12.2 --- Hybridization --- p.43 / Chapter 2.12.3 --- Antibody incubation --- p.43 / Chapter 2.12.4 --- Histochemistry --- p.44 / Chapter Chapter 3 --- Results --- p.46 / Chapter 3.1 --- The molecular cloning of ECE-2 from rat brain --- p.46 / Chapter 3.2 --- Sequence characteristics of rat ECE-2 --- p.52 / Chapter 3.3 --- Comparison of rat ECE-2 with bovine and human ECE-2 and with the rat ECE-1 --- p.53 / Chapter 3.4 --- Tissue distribution of ECE-2 in rat and localization in C6 glial cells by RT-PCR --- p.60 / Chapter 3.5 --- ECE-2 in rat embryos at different gestation stages by RT-PCR --- p.60 / Chapter 3.6 --- ECE-2 distribution in C6 glioma cells --- p.63 / Chapter 3.7 --- ECE-2 distribution in rat embryo E15.5 --- p.63 / Chapter 3.8 --- ECE-2 distribution in rat brain sections --- p.63 / Chapter Chapter 4 --- p.74 / Discussion --- p.74 / References --- p.81