The disribution of ganglioside sialidase on developing mouse tooth germ

神经节苷脂唾液酸酶在小鼠牙胚发育中的分布

• 批准号: 13671892
• 负责人: AKITA Hirotoshi
• 金额: $ 1.79万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671892/
• 关键词: ganglioside sialidase; tooth germ; transgenic mouse; immunocytochemistry; histology; tissue fixation; 歯胚上皮; 免疫組織化学; 透過型電子顕微鏡; 歯胚・上皮

Ganglioside sialidase is one type of the animal sialidases, whose activity is innately localized on cell membrane by biological definition. This project aimed at confirming the morphological localization and biological role of this enzyme. An immunocytochemical study proved the enzyme being localized on cell membrane of COS cells transfected with bovine cDNA of the enzyme, and of skeletal muscle cells in transgenic mice expressed with the human cDNA. To elucidate a biological role of the enzyme, a histological study was done in incisors of abovementioned transgenic mice. Mouse incisors continue to grow during the animal life. Therefore we can study an effect of some experimental condition on tooth formation at any time of the animal life. The histological changes of the enamel organ and tooth were ambiguous in the transgenic mice because of inadequate tissue fixation with paraformaldehyde. To get good histological features, the tissue fixation method needed to be improved. Mice under anesthetization were fixed with the chemical fixative which was infused into the left ventricle through a catheter by using a peristaltic pump. The fixative circulating the body along blood vessels was drained from the right atrium cut open. The shorter surgical operation time to insert a catheter into the left. ventricle you will take, the better histological features you will get. A catheter was devised for shortening the surgical operation time. The best catheter was a polyethylene tubing 0.9mm in diameter connected to a metal needle (26 gauge) at the one end. The left ventricle was punctured by the needle after thoracotomy. To hold the needle in place the heart and needle were glued by using Aron Alpha A (Toagosei Co., Ltd.), one of surgical glues. Taking much time for improving the tissue fixation method, the histological study has not yet been completed.

神经节苷脂唾液酸酶是动物唾液酸酶的一种类型，其活性天生地定位于生物学定义上。该项目旨在确认该酶的形态定位和生物学作用。一项免疫细胞化学研究证明了该酶位于用酶的牛cos染色的COS细胞的细胞膜上，以及用人cDNA表达的转基因小鼠中的骨骼肌细胞的骨骼肌细胞。为了阐明酶的生物学作用，在上述转基因小鼠的切牙中进行了组织学研究。小鼠门牙在动物生活中继续增长。因此，我们可以在动物寿命的任何时候研究某些实验条件对牙齿形成的影响。由于用多聚甲醛的组织固定不足，在转基因小鼠中，搪瓷器官和牙齿的组织学变化是模棱两可的。为了获得良好的组织学特征，需要改进组织固定方法。在麻醉下的小鼠用化学固定剂固定，该化学固定剂通过使用蠕动泵通过导管注入左心室。将沿血管循环的固定剂从右心庭切开的开放处排出。较短的手术操作时间将导管插入左侧。您将采用的心室，将获得更好的组织学特征。设计了用于缩短外科手术时间的导管。最好的导管是直径为0.9mm的聚乙烯管，一端连接到金属针（26量规）。胸部切开术后左心室用针刺。为了将针头固定在适当的位置，使用Aron Alpha A（Toagosei Co.，Ltd.）胶合了心脏和针头。组织学研究花了很多时间来改善组织固定方法，尚未完成。