碩士 / 國立陽明大學 / 生理學研究所 / 93 / Muscarinic acetylcholine receptors (mAChRs) are G-protein-coupled receptors with seven putative transmembrane domains. They play important roles in neuronal and non-neuronal functions, such as cell proliferation, differentiation, apoptosis, etc. In the past decades, zebrafish has become a model organism to study vertebrate development. We use the zebrafish as model to study the possible role of M2 mAChR (M2R) in blood and blood vessel formation in embryos. Previously, we cloned and characterized the pharmacological property of the zebrafish M2R gene. We demonstrated that the M2R mediates the embryonic bradycardia induced by acetylcholine by morpholino-modified antisense oligonucleotide (MO) gene knockdown methods. The M2-MO injected embryos also displayed major development defects (1) hematopoiesis: cardiac pale blood phenotype (2) vasculogenesis: delayed appearance of some blood vessels; (3) eye structure: small eyes, (4) swim bladder: no inflation, etc.
In this study, we demonstrated the cardiac pale blood phenotype was due to decrease in accumulation of circulatory blood cell numbers. Consistently, we found that the expression of globin αe1 and globin βe1 were suppressed in M2-MO-injected embryos. These phenotypes were M2R-specific, because the embryos injected with 4-mismatched M2-MO (M2-4mp) did not show the phenotype and the defect could be restored by M2-cRNA. The transgenic zebrafish line TG(fli1:EGFP), in which the blood vessels are marked with EGFP under the control of fli1 promoter, was utilized for blood vessel study. We demonstrated that the appearance of dorsal longitudinal anastomatic vessel was delayed in the M2-MO injected embryos. Our results suggest that M2R may play some roles in the zebrafish embryo hematopoiesis and vasculogenesis.
| Identifer | oai:union.ndltd.org:TW/093YM005116018 |
| Date | January 2005 |
| Creators | Ya-wen Hsu, 許雅雯 |
| Contributors | Ching-Fong Liao, 廖欽峰 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 90 |
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