We have studied a series of 22 human medullary carcinomas (MCTs), both primary and metastatic, using immunocytochemistry (ICC) to localize calcitonin and somatostatin peptide and in situ hybridization (ISH) to localize calcitonin and somatostatin mRNA. All tumors were positive for calcitonin peptide with ICC, which often showed considerable intercellular heterogeneity, with many cells having undetectable levels of calcitonin. However, calcitonin mRNA localized by ISH was much more uniformly distributed, indicating that MCT tumor cells may retain the capacity to both synthesize and store calcitonin, whereas others lose their storage but not their synthetic capacity. Somatostatin peptide and mRNA were found in tumors from 15 patients. In contrast to the pattern seen with calcitonin, somatostatin mRNA and peptide were usually found in single scattered cells. When correlation was possible, the same cell showed positivity for somatostatin mRNA on ISH and positivity for somatostatin peptide on ICC. However, in one tumor many more cells were positive for mRNA than for peptide, suggesting that only a proportion of cells retained the ability to store the peptide. The variation in cellular content of immunoreactive calcitonin is interpreted as resulting from either an increased tumor growth rate or reduced ability to store peptide in a less differentiated tumor. With somatostatin there was good correlation between mRNA and peptide content, but it occurred in single widely scattered cells, most tumor cells being negative for both peptide and mRNA. It is suggested that somatostatin production might be associated with a reduction in the growth of the cell concerned, either through a differentiation step or through a direct effect of the hormone.(ABSTRACT TRUNCATED AT 250 WORDS)