Detection of mRNAs containing regulatory peptide coding sequences using synthetic oligodeoxynucleotides

Abstract

To understand the regulation of the production of peptide hormones, it is vital to elucidate their biosynthetic pathways. We chose to study a major regulatory peptide, vasoactive intestinal peptide (VIP), a peptide possessing both neurotransmitter and neurohormone actions. To identify the specific peptide mRNA we are using, as hybridization probes, radiolabeled synthetic oligodcoxynucleotides with sequence complementary to the predicted peptide mRNA sequence. Employing this approach, we identified and partially purified a ∼ 1600‐base long mRNA containing VIP related sequences which can be translated in vitro into VIP‐immunoreactive polypeptides. Such mRNA was detected in normal VIP producing tissue (rat brain), as well as in a tumor producing VIP (human buccal tumor). This mRNA differs in size from a known VIP‐mRNA identified in human neuro‐blastoma cells, suggesting the possibility of different VIP‐mRNAs in different cell types.