摘要
本研究之目的在於發展出量化並可透過簡易計算神經軸突生長長度的方法。希望能應用於快速檢測外在環境分子，例如：細胞外基質 (extracellular matrix) 及細胞黏附分子 (cell adhesion molecules) 對神經細胞生長的研究，期待能進一步瞭解外在環境分子促進神經生長的訊號傳遞路徑。
本實驗研究利用微接觸壓印法 (micro-contact printing) ，使待測外在環境分子轉印在玻片上，讓軸突沿著微米級圖案上生長，再用PDMS 模版 (stencil) 侷限海馬迴神經細胞的貼附，能有效檢測外在環境分子對軸突的刺激而影響其生長情形。本實驗選用neuroligin-1做為刺激神經細胞的外在環境分子，並加入結合微接觸壓印法及PDMS 模版侷限的系統，觀察軸突的生長情形。本研究之計算方式與免疫螢光染色法之傳統計算方式不同，可在同一條神經細胞上觀察數天的生長情況，且輕易計算出軸突生長長度。此結果發現neuroligin-1能影響軸突的生長。故進一步與神經細胞生長在laminin、poly-L-lysine及collagen I對酪胺酸磷酸化蛋白作程度上的比較，期許能尋找出neuroligin-1 促進軸突生長的路徑，但結果顯示在不同的環境下生長的神經細胞，用銀染的方式或是西方點墨法，酪胺酸磷酸化蛋白之間的磷酸化程度並沒有什麼太大的差異。希望利用此方法學，能建立快速且有效的量化軸突生長長度之方法，未來可望能對促進神經生長方面有所貢獻。

Abstract
The purpose of my study was to develop a convenient method for quantifying the growth rate the axons. This method would also be applied to study how extrinsic molecules, such as extracellular matrix molecules (ECM) and cell adhesion molecules (CAM), affected the growth of axons and to further investigate the cellular signaling paths that might involve in axonal growth.
Here, we made the CAM-coated fine lines on the surface of glass coverslips for guiding the growth of axons by micro-contact printing technique. The cell bodies of dissociated rat hippocampal neurons were confined to a designated area of the same coverslip by using a stencil made from Polydimethylsinloxane (PDMS). By using this setup, I could make continual observations of the growth of the same sets of axons over periods of many days. By using this method, I found that neuroligin-1, a postsynaptically residing CAM, could enhance the growth of neuronal axons. By using a conventional method, which involved the quantifying the lengths of axons as processes positively fluorescence immunostained by an axon marker, neuroligin-1 and laminin, but not collagen was found enhance the axonal growth. The results indicate that the methodology developed herein will be a useful tool for neuroscientists while studying the issues related to the growth of axons in the future.
Furthermore, I started to investigate the phosphotyrosine levels of the proteins of cultured rat hippocampal neuron grown on glass coverslip with its surface coated with laminin, poly-L-lysine or collagen. The results indicated that neurons growing on the surface coated with these different molecules exhibited a similar set of proteins, as indicated by silver-stained SDS-PAGE, and nearly identical phosphotyrosine protein patterns, as indicated by Western blotting analysis. It will be of interest in the future if such studies could be conducted on isolated axons and at different times after axons are exposed to these extracellular molecules.

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