Molecular structure,spatiotemporal expression and biological functions of zebrafish muscle regulatory factors MRF4a and MRF4b

• Main Authors: Chun-Kai Li, 李駿凱
• Other Authors: 陳曜鴻
• Format: Others
• Language: zh-TW
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/58491480731283605399

碩士 === 淡江大學 === 生命科學研究所碩士班 === 95 === Our previous study shows that zebrafish has two MRF4 isoforms, MRF4 a and MRF4 b. The two MRF4 proteins exhibited a 97 % identity rate. To detect the spatiotemporal expression pattern of mrf4 a and mrf4 b at mRNA level, we used in situ hybridization. Our data showed mrf4 a expressed in trigeminal nerve, olfactory placode and RB (Rohon-beard) neuron. mrf4 b expressed in somite. To test the biological function of each mrf4, we first used anti-sense morpholino to knockdown mrf4 a and mrf4 b individually.Curved body were observed after knockdown. Then we used muscle markers F59, α-actin、tnnt1、tnnt3b，and neuron markers aat (anti-acetylated tubulin), Znp1, α-bungarotoxin, Zn5, Zn12 to detect morphological change of mrf4 morphant in skeletal muscle, slow muscle, fast muscle, motor neuron, innervations, acetylcholine receptor cluster and RB neuron. Our data shows mrf4 a knockdown led to RB neuron dendrite loss, motor neuron innervation, mrf4 b knockdown led to misalignment of slow muscle, α-actin, tnnt1, tnnt3b down reglation, secondary motorneuron defect. On the basis of our observations, we suggest that mrf4 a expression is involved in neuron development, mrf4 b expression is involved in muscle development.