神經壞死病NNV，為發生於石斑魚魚苗階段重要的疾病之一。主要的病症發生於中樞神經系統及視網膜。在現場的觀察中，發現此一病毒在青斑魚白身階段以後，其致病力有下降的趨勢。推測病毒的感染可能與魚類神經系統的發育有關。但要研究此一相關性，則需要相關神經發育因子的幫助，以界定神經發育的狀況。生長相關蛋白-43 (GAP-43) 是在神經發育與再生時在軸突生長錐會高度表現的一種蛋白，其功能為神經元軸突延伸之誘導因子。由於其表現特性，因此在哺乳動物與兩棲類動物的研究中，生長相關蛋白-43常作為評估個體神經元之發育與再生的生物標記。在先前試驗中已選殖到點帶石斑魚的生長相關蛋白-43，且證明其可以作為神經元之發育及再生的生物標記。並利用視神經損傷的方式證明可誘使生長相關蛋白-43高度表現，並增加NNV病毒在視神經的表現。但由於NNV病毒主要侵襲腦部組織。故本研究將進一步探討生長相關蛋白-43與NNV病毒間可能的關係。我們利用外科手術方式對魚隻腦部進行創傷，發現其生長相關蛋白-43的表現量有顯著的增加，表示創傷可以誘導腦部神經的再生。在此一再生狀態時以神經壞死病毒攻擊，其腦部之病毒量也有顯著的提升。在免疫組織染色的觀察上，創傷部位其生長相關蛋白-43蛋白具有高度表現，而其表現的位置與NNV病毒感染細胞位置一致。此一結果顯示，生長相關蛋白-43確實在創傷的腦區有較高量的表現，這些高量表現的細胞較易受到病毒感染。這些結果顯示，NNV的感染與腦細胞的發育再生有很明顯的關聯。此外我們在篩選與NNV外鞘蛋白互動的宿主蛋白時發現肌動蛋白纖維與生長相關蛋白-43蛋白具有相關性。且利用藥物的投與，降低石斑魚魚鰭細胞株GF-1細胞內肌動蛋白纖維的組裝，不論是在細胞內或被釋放到培養基的病毒數則有明顯的下降。因此我們推論，肌動蛋白纖維的組裝應與病毒感染早期複製有關。但其詳細的機轉目前仍在釐清當中。

Nerve necrotic virus (NNV) was a serious pathogen in grouper fry stage. The major attack site was the central nerve system (CNS) and retina. In the observation in field, the mortality of this disease was dramatically decreased in the juvenile stage. It is deduced the NNV infection is highly related with the development and regeneration of CNS. But the CNS development biomarker is essential for the related research. Growth associated ptotein-43 (GAP-43) is a highly expressed protein during neuron developing and regenerating of axonal growth cone. In fact, the function of GAP-43 is one of the inducible factors which can extend axonal growth cone. Hence, GAP-43 is already recognized as a fabulous biomarker to evaluate the individual neuron development and regeneration in mammalian model. According to previous studies researched in our laboratory, GAP-43 used as a biomarker of neuron development and regeneration in Epinephelus coioides. And we found the mRNA of GAP-43 was elicited while the optic nerve was transected, and the NNV gene expression level was elevated while the nerve necrotic virus (NNV) was challenged. Since brain was also the major attack site of NNV, the same phenomenon of GAP-43 should be proven in brain. We create the trauma of fish brain using surgery method, and the express of GAP-43 was evaluated. The results shown the expression level of GAP-43 could also up regulated. Meanwhile, the expression NNV level of are up regulated in brain while NNV is challenged, and the NNV amount is increase after challenge in the brain trauma fish. In IHC staining, the GAP-43 was expressed in the injury brain around the puncture point, and the high GAP-43 expressed cell is suspected for NNV in the challenge fish. These revealed the development and regeneration status of brain was related with the NNV infection. Meanwhile, we select some proteins which can interact with NNV coat protein in host cell and discover the NNV coat protein can interact with actin. Than, we use the medicine inhibited assembly of actin filament to GF-1 cell, and discover amount of NNV particle in cell body or in medium are decrease in the treatment group. So, we think that it have a relationship between assemblely of actin filament and NNV replication. But the detail mechanism still need clarify in the future.

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