Organotypic rat cerebellar slice culture as a model to analyze the molecular pharmacology of GABAA receptors
Organotypic rat cerebellar slice culture as a model to analyze the molecular pharmacology of GABAA receptors
复制标题
DOI：
10.1016/s0924-977x(02)00024-x
复制
发表时间：
2002-06-01
影响因子：
5.6
通讯作者：
Lüddens, H
中科院分区：
文献类型：
--
作者：
Davids, E;Hevers, W;Lüddens, H
在大鼠小脑切片培养物中保存神经元回路为监测GABA A受体亚型的发展和表征其药理学提供了优势。将8-11日龄的Sprague-Dawley大鼠斩首。将它们的小脑切成400 μ m的薄片并转移到培养皿中。培养物的细胞活力和器官型小脑组织保持良好保存长达3周。在这些先进的培养技术中引入了放射自显影程序，并在不存在和存在10 μ M地西泮的情况下使用[H-3]Ro 15-4513分别显示所有苯二氮卓类（BZD）和地西泮不敏感（DIS）结合位点。由于GABA（A）/BZD受体α 6亚基变体的表达仅限于小脑颗粒细胞，且BZD受体激动剂地西泮对该亚基的亲和力非常低，因此培养期间DIS [H-3]Ro 15-4513结合位点的变化可归因于α 6亚基表达的变化。在培养物中观察到总的和DIS [H-3]Ro 15-4513结合位点的时间依赖性发展，在第一周期间，GABA受体α 6亚基水平有增加的趋势。这些结果表明，用组织块制备的小脑脑片可用于观察小脑脑片的形态学变化，并可用于研究GABA（A）/BZD选择性药物对大鼠小脑GABA（A）受体生后发育的药理作用。(C)2002 Elsevier Science B. V.保留所有权利。
The preservation of the neuronal circuitry in rat cerebellar slice cultures provides an advantage in monitoring the development and characterizing the pharmacology of GABA A receptor subtypes. Sprague-Dawley rats, 8-11 days of age, were decapitated. their cerebella were cut into 400-mum slices and transferred into culture dishes. Cell viability and organotypic cerebellar organization of the culture remained well preserved up to 3 weeks. Autoradiographic Procedures were introduced in these advanced culture technique and employed [H-3]Ro 15-4513 in the absence and presence of 10 muM diazepam to visualize all benzodiazepine (BZD) and diazepam-insensitive (DIS) binding sites, respectively. Since expression of the alpha6 subunit variant of the GABA(A)/BZD receptor is restricted to the cerebellar granule cells and the BZD receptor agonist diazepam has very low affinity for this subunit, changes in DIS [H-3]Ro 15-4513 binding sites during cultivation time can be attributed to changes in alpha6 subunit expression. A time-dependent development of total and DIS [H-3]Ro 15-4513 binding sites were observed in the culture with a trend towards an increase in GABA, receptor alpha6 subunit levels during the first week. These findings suggest that explant preparations can be used to examine morphological changes in rat cerebellar slices, In addition, these preparations can be utilized to study the pharmacological effects of GABA(A)/BZD selective drugs on postnatal development of GABA(A) receptors in rat cerebellum. (C) 2002 Elsevier Science B.V. All rights reserved.