In situ hybridization with cDNA probes: Its application to the detection of mRNAs for myelin proteins (PLP and MBP) in mouse brain sections.

Abstract

In situ hybridization is a method for identifying specific DNA or RNA sequences in cells or tissues. It was originally developed for the detection of specific DNA sequences in chromosomes (2, 6), then extended to the detection of specific endogenous mRNA in cells (5). Various kinds of nucleotide sequences, such as complementary DNA (cDNA), antisense RNA and synthetic oligonucleotide that have been labeled with radioactive or otherwise detectable molecules, could be acceptable as probes for in situ RNA hybridization, as well as Northern hybridization. Among these probes, the radioactive cDNA probe is the most widely used for in situ hybridization. With a rapidly increasing number of cDNAs corresponding to molecules of interest, in situ hybridization has been extensively used in various fields such as neurobiology, endocrinology, developmental biology, etc. The detection of mRNAs for myelin basic protein (MBP) and myelin proteolipid protein (PLP) (7, 12, 13) is a good example of successful application of in situ hybridization. Here we describe our evaluation of some technical conditions which are generally used in in situ hybridization. We also show the distribution of MBP and PLP mRNAs in the mouse brain sections revealed by the relatively simple but effective method of in situ hybridization.