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使用共聚焦扫描光学显微镜分析检测电鳗正常和去神经支配电细胞中的胆碱乙酰转移酶。

Choline acetyltransferase detection in normal and denervated electrocyte from Electrophorus electricus (L.) using a confocal scanning optical microscopy analysis.

作者信息

Nunes-Tavares N, Cunha-E-Silva N L, Hassón-Voloch A

机构信息

Laboratório de Físico-Química Biológica, Centro de Ciências da Saude, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, 21491-590, Brasil.

出版信息

An Acad Bras Cienc. 2000 Sep;72(3):331-40. doi: 10.1590/s0001-37652000000300007.

Abstract

Acetylcholine is the neurotransmitter responsible for the transmission of impulses from cholinergic neurons to cells of innervated tissues. Its biosynthesis is catalyzed by the enzyme Choline acetyltransferase that is considered to be a phenotypically specific marker for cholinergic system. It is well known that the regulation of Choline acetyltransferase activity under physiological and pathological conditions is important for development and neuronal activities of cholinergic functions. We observed the distribution of Choline acetyltransferase in sections from the normal and denervated main electric organ sections of Electrophorus electricus (L.) by immunofluorescence using a anti-Choline acetyltransferase antibody. The animals were submitted to a surgical procedure to remove about 20 nerves and after 30 and 60 days, they were sacrificed. After 30 days, the results from immunohistochemistry demonstrated an increase on the Choline acetyltransferase distribution at denervated tissue sections when compared with the sections from the normal contralateral organ. A very similar labeling was observed between normal and denervated tissue sections of the animals after 60 days. However, Choline acetyltransferase activity (nmolesACh/ min/ mg of protein) in extracts obtained from electrocyte microsomal preparation, estimated by Fonnun's method (Fonnun 1975), was 70% lower in the denervated extracts.

摘要

乙酰胆碱是负责将冲动从胆碱能神经元传递到受支配组织细胞的神经递质。其生物合成由胆碱乙酰转移酶催化,该酶被认为是胆碱能系统的表型特异性标志物。众所周知,在生理和病理条件下,胆碱乙酰转移酶活性的调节对于胆碱能功能的发育和神经元活动很重要。我们使用抗胆碱乙酰转移酶抗体通过免疫荧光观察了电鳗正常和去神经支配的主电器官切片中胆碱乙酰转移酶的分布。对动物进行手术切除约20条神经,30天和60天后将它们处死。30天后,免疫组织化学结果表明与正常对侧器官的切片相比,去神经支配组织切片中胆碱乙酰转移酶的分布增加。60天后在动物正常和去神经支配的组织切片之间观察到非常相似的标记物。然而,通过Fonnun方法(Fonnun,1975年)估计,从电细胞微粒体制备物中获得的提取物中的胆碱乙酰转移酶活性(每毫克蛋白质每分钟纳摩尔乙酰胆碱)在去神经支配的提取物中低70%。

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