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首页> 外文期刊>Zoological Science >Characterization and Expression of Trypsinogen and Trypsin in Medaka Testis
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Characterization and Expression of Trypsinogen and Trypsin in Medaka Testis

机译：Medaka睾丸中胰蛋白酶原和胰蛋白酶的表征和表达

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Previously, we reported that the medaka testis abundantly expresses the mRNA for trypsinogen, which is a well-known pancreatic proenzyme that is secreted into and activated in the intestine. Currently, we report our characterization of the medaka trypsin using a recombinant enzyme and show that this protein is a serine protease that shares properties with trypsins from other species. Two polypeptides (28- and 26-kDa) were detected in the testis extracts by Western blot analysis using antibodies that are specific for medaka trypsinogen. The 28-kDa polypeptide was shown to be trypsinogen (inactive precursor), and the 26-kDa polypeptide was shown to be trypsin (active protease). We did not detect enteropeptidase, which is the specific activator of trypsinogen, in the testis extract. Immunohistochemical analyses using the same trypsinogen-specific antibody produced a strong signal in the spermatogonia and spermatozoa of the mature medaka testis. Substantial staining was found with spermatocytes, whereas extremely weak signals were observed with spermatids. In vitro incubation of testis fragments with the trypsinogen antibody strongly inhibited the release of sperm from the testis into the medium. Trypsin activity was detected in sperm extracts using gelatin zymographic analysis. Immunocytochemistry showed that trypsinogen and trypsin were localized to the cell membranes surrounding the sperm head. Collectively, these results suggest that trypsin plays an important role in the testis function of the medaka.
机译：先前，我们报道了med高睾丸大量表达胰蛋白酶原的mRNA，胰蛋白酶原是一种众所周知的胰腺酶，被分泌到肠中并在肠中被激活。目前，我们报告了我们使用重组酶对中aka胰蛋白酶的鉴定，并表明该蛋白是一种丝氨酸蛋白酶，与其他物种的胰蛋白酶具有相同的特性。使用对Medaka胰蛋白酶原特异的抗体，通过Western印迹分析在睾丸提取物中检测到两种多肽（28-kDa和26-kDa）。 28-kDa多肽显示为胰蛋白酶原（无活性的前体），而26-kDa多肽显示为胰蛋白酶（活性蛋白酶）。我们未在睾丸提取物中检测到肠肽酶，肠肽酶是胰蛋白酶原的特异性激活剂。使用相同的胰蛋白酶原特异性抗体的免疫组织化学分析在成熟的高aka睾丸的精原细胞和精子中产生了强信号。发现精细胞明显染色，而精细胞则观察到极弱的信号。睾丸片段与胰蛋白酶原抗体的体外温育强烈抑制了精子从睾丸向培养基中的释放。使用明胶酶谱分析检测精子提取物中的胰蛋白酶活性。免疫细胞化学显示，胰蛋白酶原和胰蛋白酶位于精子头部周围的细胞膜上。总的来说，这些结果表明胰蛋白酶在青aka的睾丸功能中起重要作用。

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