权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

SYNAPTIC FORMATION IN CUTURED SENSORY

精细感觉中的突触形成

基本信息

批准号：
06640887
负责人：
NAGAI Takatoshi
金额：
$ 1.47万
依托单位：
TEIKYO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06640887/
关键词：
Culture Linfual epithelium Glossopharyngeal nerve Amphibian Teste Synapse

项目摘要

The project developed a basic technique to maintain lingual epithelium in culture, which allows us to study how synapses are made between gustatory nerve fibers and taste receptor cells. The glossophryngeal nerve ganglion and the lingual epithelium of the Mexican salamander, axolotl (ambystoma mexicanum) , were used. Neurons were isolated from the ganglion with enzymatic treatment and placed in L-15 culture medium. In 24-48 hours the neurons in culture started to extend processes, which grew into 1 mil meter in length by 2 weeks. Small pieces of the lingual epithelium containing taste buds were dissected out and kept in the medium. In one week the epithelial cells attached to the bottom of a culture dish and grew well. However, the taste bud cells lost a rosette-like arrangement distinct from general epithelial cells. Outside of the taste buds the glossopharyngeal nerve makes synapses with a few solitary cells. Electron-microscopic study revealed that those cells are of mechanosensory function. To maintain the taste bud cells of three-dimensional conformation pieces of the lingual epithelium were embedded in collagen gel, immersed in L-15 medium and kept at room temperature. The taste buds did not survive more than 24 hours, possibly due to paucity of oxygen. When the general epithelial cells were stripped off by collagenase, the taste buds were maintained for about 6 days. CO_2-incubator is required to achieve optimal temperature and sufficient oxygen for culturing the taste buds.

该项目开发了一种基本的技术，以保持舌上皮细胞的文化，这使我们能够研究如何突触之间的味觉神经纤维和味觉受体细胞。舌咽神经节和舌上皮的墨西哥蝾螈，蝾螈（ambystoma mexicanum），使用。用酶处理从神经节分离神经元，并置于L-15培养基中。在24-48小时内，培养物中的神经元开始延伸突起，其在2周时长到1密耳米长。切下含有味蕾的舌上皮小片并保存在培养基中。一周后上皮细胞贴壁生长良好。然而，味蕾细胞失去了玫瑰花状排列不同于一般的上皮细胞。在味蕾外，舌咽神经与一些孤立的细胞形成突触。电镜观察显示，这些细胞具有机械感觉功能。为了保持味蕾细胞的三维构象，将舌上皮切片包埋在胶原凝胶中，浸入L-15培养基中并在室温下保存。味蕾存活不超过24小时，可能是由于缺氧。当用胶原酶剥离一般上皮细胞时，味蕾保持约6天。CO_2培养箱需要达到最佳的温度和足够的氧气来培养味蕾。