Analysis of gene expression in neurons and synapses by multi-color in situ hybridization

Abstract

In situ hybridization is a powerful and highly specific method to reveal cell and tissue specific gene expression. It is especially critical for neuroscience applications where the enormous heterogeneity of cells as well as tissue composition present a significant methodological and conceptual challenge. Here we present two protocols that allow efficient visualization of transcripts of interests from animals with high variety in their neuronal tissue density and composition with molluscs, arthropods, and ctenophores as illustrative examples. The entire protocol takes up to 4 days. It was successfully applied both for multi-color detection of neuronal and synaptic transcripts in individually identified neurons of Aplysia californica , a powerful model in analysis of cellular bases of behavior and memory.