小白鼠貯精囊自體中胰蛋白?抑制因數的生理研究

Abstract

前人之研究而知，從小白鼠貯精囊分泌液純化不含醣鏈的胰蛋白?抑制因數（Irypsin Inhibitor，簡稱TI）屬Kazal-type proteinase inhibitor，其分子量為6.4 kDa，只存在雄性生殖系統中貯精囊、前列腺及凝固腺（coagulating gland）等部位。探討TI對精蟲活性的影響為本論文之研究重點，使用間接螢光檢定法及直接的組織染色法來觀察TI結合在精蟲上的位置，可以發現TI會結合在精蟲頭端的頂體（acrosome）位置。由chlortetracycline （CTC）螢光染色分析和Coomassie Blue （CB）染色的方法研究而知，TI可抑制由牛血清白蛋白（bovine serum albumin）所誘發精細胞產生獲能作用（capacitation）及頂體反應（acrosome reaction）的能力。此外，使用冷熱的方式（frozen-thawed method）所得到溶解性的精蟲蛋白，通過TI親和性管柱，可得一個分子量約為15kDa的蛋白，而此蛋白可能為TI在精細胞上的結合蛋白。

A kazal-type trypsin inhibitor (TI) has been previously purified from seminal vesicle secretion in our laboratory. The protein contains no carbohydrate and its molecular weight is 6.4 kDa as estimated by SDS-electrophoresis. Among the reproductive tracts of mice, TI is product only male accessory sexual gland such as seminal vesicles, prostate and coagulating gland. This work aimed to assess the effect of TI on sperm activity. Moreover, TI could suppressed the bovine serum albumin induced sperm capacitation and acrosome reaction as assayed by the chlortetracycline (CTC) fluorescence method and the Coomassie Blue (CB) staining technique. A protein with a Mr around 15 kDa could be isolated from the supernatant of frozen-thawed epididymal sperm suspension through an affinity column of agrose linked with TI.