The Secretory Granule and Pro-opiomelanocortin Processing inXenopusMelanotrope Cells During Background Adaptation

Abstract

In this immunocytochemical study, we used light and electron microscopic observations in combination with morphometry to analyze the processing of pro-opiomelanocortin (POMC) in melanotrope cells of the intermediate pituitary of Xenopus laevis adapted to either a white or a black background. An antiserum was raised against a synthetic peptide including the cleavage site between ACTH and β-lipotropic hormone in Xenopus. Western blotting revealed that this antiserum recognizes only a 38-kD protein, the POMC prohormone, from extracts of Xenopus neurointermediate pituitary. Light immunocytochemistry showed differential immunostaining for anti-POMC compared to anti-α-MSH. Anti-POMC was predominantly found in the perinuclear region, whereas anti-α-MSH yielded staining throughout the cytoplasm. Immunogold double labeling revealed that electron-dense secretory granules (DGs) show high immunoreactivity for anti-POMC and low immunoreactivity for anti-α-MSH. Electron-lucent granules (LGs) are immunoreactive to anti-α-MSH only. Moderately electron-dense granules (MGs) revealed intermediate reactivity compared to DGs and LGs. Background light intensity has significant effects on the morphology and the immunoreactivity of the secretory granules. Black-adapted animals have 4.5 times as many DGs and MGs as white-adapted animals. In addition, the MGs in black animals show 42% more anti-α-MSH immunogold than the MGs in white animals. Together, these findings indicate that the three granule types represent subsequent stages in granule maturation. Adaptation to a black background stimulates the formation of young immature granules, while at the same time the processing rate during granule maturation increases.