We present a new method for the simultaneous detection of two mRNA species within individual neurons. The technique involves the use of radio-labeled and digoxigenin-labeled cRNA probes, the application of which confers a high specificity and sensitivity to the in situ hybridization analysis. We demonstrate the use of this method by illustrating the coexpression of preprogonadotropin-releasing hormone (GnRH) mRNA and preprogalanin mRNA in neurons in the rat forebrain and report a distinct sexual dimorphism in galanin gene expression by GnRH neurons. Coupling this technology with semi-quantitative analysis of the mRNA species hybridized with the isotopically labeled mRNA would permit studies of gene regulation in individual cells among the heterogeneous populations of the brain.
ASJC Scopus subject areas
- Molecular Biology
- Cellular and Molecular Neuroscience
- Cell Biology