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研究生:陳振國
研究生(外文):Jen-Guo Chen
論文名稱:斑馬魚鈉離子通道Nav1.6亞型在運動及軸突投射上的功能研究
論文名稱(外文):The role of zebrafish sodium channel Nav1.6 on locomotion and axonal projection
指導教授:蔡惠珍宋晏仁宋晏仁引用關係
指導教授(外文):Huey-Jen TsayYen-Jen Sung
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:神經科學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:英文
論文頁數:94
中文關鍵詞:鈉離子通道1.6亞型運動行為神經網路軸突投射細胞程序性死亡
外文關鍵詞:Nav1.6locomotionaxonal projectionprogrammed cell death
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中文摘要
電位感應式鈉離子通道負責引發軸突與樹突的動作電位,在神經系統信息傳遞中扮演重要的角色。鈉離子通道亞型1.6集中在有髓鞘包覆軸突上的蘭氏節中,是出生後小鼠脊椎內動作神經元的主要鈉離子流貢獻者。帶著鈉離子通道亞型1.6基因變異的med小鼠會產生漸進式神經與肌肉系統衰竭的運動末稍板症,而神經鈉離子流減少的斑馬魚mao變異種表現觸覺反應缺失與較少的Rohon-Beard (RB) 神經元死亡。這顯示電流活動在神經存活與軸突投射上扮演關鍵的角色。
本論文將研究斑馬魚鈉離子通道亞型1.6在運動、神經存活與軸突投射上扮演的功能。斑馬魚鈉離子通道1.6亞型的反義morpholino寡聚核苷酸(AMO)被用來抑制在發育過程中鈉離子通道1.6亞型的表現。AMO注射的胚胎在絨毛膜中的擺尾行為、因觸碰而引發的擺尾行為與在渦流中保持平衡的游泳能力都出現缺失。而AMO注射組與注射scramble序列的morpholino寡聚核苷酸控制組,無論是在體節數與肌肉結構上都無明顯差異,顯示AMO組的動作缺失並非因發育遲緩造成。
造成AMO注射組的運動缺失至少有兩種可能,第一,神經網路、運動神經肌肉接點以及肌肉本身結構都正常,但是因為鈉離子通道1.6蛋白的表現量減少而影響了電訊號傳導;第二,部分的運動神經
及肌肉在結構上產生改變,造成運動功能缺失。為了區辨這兩種可能,我們檢驗了神經網路軸突投射、神經肌肉接點與肌肉排列。研究結論是鈉離子通道1.6亞型在三種運動行為上扮演重要的角色,但在AMO注射的斑馬魚中沒發現有明顯初級及二級運動神經元投射異常與肌肉排列異常的情形,而電流活性可以調控RB神經元細胞程序性死亡及其週邊樹突的投射範圍。
Abstract
Voltage-gated sodium channels play critical roles for the electrical signaling in the nervous system by transmitting action potentials in axons and dendrites. Nav1.6, which located at the nod of Ranvier of myelinated neurons, is the major sodium influx contributor in postnatal spinal motoneurons. Previous studies indicated that med mutant mice harboring loss-function of Nav1.6 had progressive neuromuscular failure (motor end plate disease). The phenotypes of zebrafish mao mutant with reduced sodium current include defect on tactile response and less cell death of Rohon-Beard neurons (RB neurons). It is suggested that electrical activity plays critical roles in neuronal survival and axonal projection. The specific role of zebrafish Nav1.6 on locomotion, neuronal survival, and axonal projection will be investigated in this study. Antisense morpholino oligonucleotides (AMO) against zebrafish Nav1.6 was used to knockdown the expression of Nav1.6 during the development. Three type of locomotion including chorion contraction, touch-evoked contraction, and the ability of swimming against swirl were examined. Embryos with Nav1.6 knockdown displayed defects on three types of locomotion. There was no significant difference in number of somite or muscle structure between scramble MO-injected embryos (SMO, as negative control) and AMO-injected embryos (no-movement AMO), suggesting that the lack of locomotion in no-movement AMO might not due to general effect of developmental delay. There are two possible reasons for locomotion phenotype in AMO group. (1) All the neural networks, neuromuscular junction, and muscle
involved in locomotion are structurally intact but lack of action potential transmission due to a reduced level of Nav1.6 sodium channel. (2) Part of the neuromuscular circuit is structurally altered and leads to functional defect. To distinguish these two possibilities, axonal projection of neural network involved in locomotion, neuromuscular junction, and muscle were examined. Our results were concluded as followed. 1. Nav1.6 played critical roles in locomotion of embryo and larvae. 2. No significant difference in motoneuron projection and muscle organization was found in no-movement AMO. 3. Electrical activity may regulate RB neuron apoptosis and RB peripheral process.
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