Combination of immunocytochemistry and in situ hybridization in the same tissue section of rat pituitary

Abstract

A procedure is described for combining avidin-biotinylated horseradish peroxidase immunocytochemistry, for localizing peptides or proteins, with in situ hybridization for localizing mRNA autoradiographically in the same cryostat section of paraformaldehyde-fixed rat pituitary. Protection against enzymatic degradation of target mRNAs during the immunocytochemical step was necessary and was accomplished by including an RNase inhibitor, 0.04% diethylpyrocarbonate, in primary and secondary antisera. This combination of methods may be useful in other tissues, as well, for (a) determining the relation of protein content to the concentration of its encoding mRNA, (b) proving the synthetic capacity of a cell in which a protein has been localized, (c) determining immunological or nucleic acid probe specificity, or (d) as an alternative to double-labeling immunocytochemical methods.