Validation of a new antiserum directed towards the synthetic c-terminus of the FOS protein in avian species:: immunological, physiological and behavioral evidence
Validation of a new antiserum directed towards the synthetic c-terminus of the FOS protein in avian species:: immunological, physiological and behavioral evidence
复制标题
DOI：
10.1016/s0165-0270(99)00067-9
复制
发表时间：
1999-09-15
影响因子：
3
通讯作者：
Berghman, LR
中科院分区：
文献类型：
--
作者：
D'Hondt, E;Vermeiren, J;Berghman, LR
在过去的10年里，对大脑中c-fos等即时早期基因表达的研究已经成为识别参与特定生理和行为功能调节的大脑区域的常用方法。由于缺乏与禽FOS蛋白交叉反应的合适抗体，这种方法在禽类中的应用受到限制。本文描述了一种针对鸡c-fos基因蛋白产物合成片段的抗体的制备。我们证明该抗体可用于多种鸟类，以研究各种药理、生理和行为刺激诱导的FOS表达。Western blot研究表明，该抗体可识别预期大小（47 kDa）的蛋白，但也可与与FOS （FOS相关抗原）序列同源的较低分子量蛋白发生一定程度的交叉反应。利用免疫细胞化学程序的不同变体，在三个不同实验室的四种鸟类中使用该抗体进行FOS免疫细胞化学。在所有病例中，抗体提供了可靠的FOS抗原鉴定。本文描述的新抗体似乎适用于几种不同鸟类和情况下FOS表达的研究。它的数量是相当大的，因此将有可能使用FOS表达作为绘制鸟类大脑活动的工具，就像多年来在哺乳动物物种中所做的那样。(C) 1999 Elsevier Science B.V.版权所有
In the past 10 years, the study of the expression of immediate early genes, such as c-fos, in the brain has become a common method for the identification of brain areas involved in the regulation of specific physiological and behavioral functions. The use of this method in avian species has been limited by the paucity of suitable antibodies that cross-react with the FOS protein in birds. We describe in this paper the preparation of an antibody directed against a synthetic fragment of the protein product of the c-fos gene in chickens (Gallus domesticus). We demonstrate that this new antibody can be used in several avian species to study FOS expression induced by a variety of pharmacological, physiological and behavioral stimuli. Western blot studies indicated that this antibody recognizes a protein of the expected size (47 kDa) but also cross reacts to some extent with proteins of lower molecular weight that share sequence homology with FOS (Fos-related antigens). FOS immunocytochemistry was performed with this antibody in four species of birds in three different laboratories utilizing diverse variants of the immunocytochemical procedure. In all cases the antibody provided a reliable identification of the FOS antigen. The new antibody described here appears to be suitable for the study of FOS expression in several different avian species and situations. It is available in substantial amounts and will therefore make it possible to use FOS expression as a tool to map brain activity in birds as has now been done for several years in mammalian species. (C) 1999 Elsevier Science B.V. All rights reserved.