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首页> 外文期刊>Brain research. Brain research protocols >Cellular localization of BM88 mRNA in paraffin-embedded rat brain sections by combined immunohistochemistry and non-radioactive in situ hybridization.
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Cellular localization of BM88 mRNA in paraffin-embedded rat brain sections by combined immunohistochemistry and non-radioactive in situ hybridization.

机译：结合免疫组织化学和非放射性原位杂交技术，将石蜡包埋的大鼠脑切片中BM88 mRNA进行细胞定位。

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摘要

With the advent of gene cloning and sequencing, it has become increasingly common to identify novel genes for which no antibody is available. The best approach to study the expression and the distribution of these new genes is by in situ hybridization. One of the challenges with this method is to define the exact cellular subtype where the gene of interest is expressed. Conventional isotopic in situ hybridization methods lack precision for cellular identification because radioactive probes often result in a scattered signal. To identify the exact cellular subtype expressing BM88, we established a rapid colocalization method using non-isotopic in situ hybridization followed by chromogenic immunohistochemistry on the same tissue section. We demonstrated that BM88, which was identified from subtractive hybridization experiments between normal and ischemic tolerant brain tissue, was expressed exclusively in neurons in normal adult rat brain. Paraffin-embedded tissue was used as it resulted in better preservation of tissue and cellular morphology, thus allowing for more accurate histological localization of gene expression. It also allowed for retrospective studies on a number of archived tissue samples.
机译：随着基因克隆和测序的出现，鉴定没有抗体可用的新基因变得越来越普遍。研究这些新基因的表达和分布的最佳方法是通过原位杂交。该方法的挑战之一是定义表达目的基因的确切细胞亚型。常规的同位素原位杂交方法缺乏细胞鉴定的准确性，因为放射性探针通常会产生散射信号。为了确定表达BM88的确切细胞亚型，我们建立了使用非同位素原位杂交，然后在同一组织切片上进行显色免疫组织化学的快速共定位方法。我们证明了BM88是通过正常和成年耐受性脑组织之间的减法杂交实验确定的，仅在正常成年大鼠脑中的神经元中表达。使用石蜡包埋的组织，因为它可以更好地保存组织和细胞形态，从而使基因表达的组织学定位更为准确。它还允许对许多已存档的组织样本进行回顾性研究。

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