In situ hybridization analysis of chemokines and chemokine receptors in the central nervous system
In situ hybridization analysis of chemokines and chemokine receptors in the central nervous system
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DOI：
10.1016/s1046-2023(02)00354-7
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发表时间：
2003-04-01
期刊：
影响因子：
4.8
通讯作者：
Streit, WJ
中科院分区：
文献类型：
--
作者：
Harrison, JK;Luo, DF;Streit, WJ
许多趋化因子和趋化因子受体的表达细胞驻留在正常和病理性中枢神经系统（CNS）组织的特点是原位杂交技术。因此，我们对这个细胞因子家族在神经生物学中的作用的理解得到了极大的提高。用于原位检测趋化因子和趋化因子受体mRNA的具体方法随着这些基因的数量而变化，这些基因已经被表征，并且包括被表征细胞特异性基因表达模式的其他研究者广泛利用的方法。我们描述的方法，我们的实验室已成功地用于表征趋化因子和趋化因子受体在中枢神经系统中的表达，专注于协议，利用放射性标记的体外转录的核糖核酸探针检测这些成绩单。因为一般的基于染料的组织学染色方法不容易区分星形胶质细胞和小胶质细胞，需要特定的免疫组化方案来确定这些胶质细胞类型的基因表达的定位。因此，包括与原位杂交程序相容的方法用于染色星形胶质细胞和小胶质细胞。(C)2003 Elsevier Science（美国）。All rights reserved.
The expression of a number of chemokines and chemokine receptors by cells resident in normal and pathological central nervous system (CNS) tissue has been characterized by in situ hybridization techniques. As a result, our understanding of the role of this cytokine family in neurobiology has been enhanced greatly. Specific methods for detecting chemokine and chemokine receptor mRNAs in situ vary with the number of these genes that have been characterized and encompass approaches widely utilized by other investigators characterizing cell-specific gene expression patterns. We describe methods that our laboratory has used successfully in characterizing chemokine and chemokine receptor expression in the CNS, focusing on protocols that utilize radiolabeled in vitro-transcribed riboprobes for detecting these transcripts. Because general dye-based histological staining methods do not readily differentiate astrocytes and microglia, specific immunohistochemical protocols are required for definitive localization of gene expression to these glial cell types. As such, methods that are compatible with the in situ hybridization procedure are included for staining astrocytes and microglia. (C) 2003 Elsevier Science (USA). All rights reserved.