In situ hybridization is a necessary experimental complement to microRNA (miRNA) expression profiling in the human brain

MicroRNAs (miRNAs) play fundamental roles in human brain neurochemistry. However, much remains to be learned in this fast-paced field. To understand how miRNAs contribute to normal biologic functions and disease states, it is critical to understand the miRNAs that are expressed in particular cell types under a range of conditions. Many tools have been developed to help describe the repertoire of miRNAs present at the tissue level in a given sample. However, tissue level miRNA profiling is inadequate to pinpoint the cellular and sub-cellular distribution of individual miRNAs. Such knowledge is especially important in the nervous system with its many cell types, microscopic heterogeneity with regard to functionally distinct cell groups, and extreme geometrical complexity in cellular shapes. We have found that in situ hybridization shows important cerebral cortical lamina-specific patterns of miRNA expression that would be lost on most tissue level expression studies, and these lamina-specific patterns can be directly relevant to human brain disease. Thus, in situ hybridization is an important experimental complement to tissue level miRNA expression profiling. Technical and theoretical aspects of this important technique are described, especially those pertinent to studying the human brain.