Previous reports have described the heterogeneity of different pituita
ry cell types on the basis of morphological and physiological criteria
. In the present study, we investigated the possible existence of dist
inct subpopulations of melanotrope cells in the intermediate lobe of t
he pituitary of the frog, Rana ridibunda. Separation of dispersed pars
intermedia cells in a Percoll density gradient made it possible to is
olate two fractions of melanotrope cells whose morphological and funct
ional properties were further characterized. Analysis of the relative
volume and number of various cellular organelles showed that high-dens
ity cells had a larger number of secretory granules than low-density c
ells. Concurrently, radioimmunoassay quantification revealed that the
concentration of alpha-melanocyte-stimulating hormone (alpha-MSH) was
2 times higher in the heavy cell population. The rate of secretion of
alpha-MSH from cultured melanotrophs was significantly higher in low-d
ensity than in high-density cells. Thyrotropin-releasing hormone (TRH)
was more potent in stimulating alpha-MSH release from the low-density
than from the high-density cell subset. In contrast, the response to
TRH persisted for a longer time in the high-density cell subpopulation
. Taken together, these data demonstrate the existence of two subpopul
ations of melanotrope cells, and indicate that the low-density cells h
ave a secretory rate substantially greater than high-density cells.