The secretory granule and pro-opiomelanocortin processing in Xenopus melanotrope cells during background adaptation.
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 beta-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-alpha-MSH. Anti-POMC was predominantly found in the perinuclear region, whereas anti-alpha-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-alpha-MSH. Electron-lucent granules (LGs) are immunoreactive to anti-alpha-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-alpha-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.
Berghs
C A
CA
Department of Cellular Animal Physiology, Nijmegen Institute for Neurosciences, University of Nijmegen, Nijmegen, The Netherlands.
Tanaka
S
S
Van Strien
F J
FJ
Kurabuchi
S
S
Roubos
E W
EW
D000222:Adaptation, Physiological / Q000502:physiology*; D000818:Animals; D015153:Blotting, Western; D003594:Cytoplasmic Granules / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D017979:Gold Colloid; D007074:Immunoglobulin G / Q000032:analysis; D007150:Immunohistochemistry; D008547:Melanophores / Q000502:physiology*; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D010902:Pituitary Gland / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D011333:Pro-Opiomelanocortin / Q000032:analysis / Q000276:immunology / Q000502:physiology*; D014982:Xenopus laevis; D000521:alpha-MSH / Q000032:analysis / Q000276:immunology
D017979:Gold Colloid; D007074:Immunoglobulin G; D000521:alpha-MSH; D011333:Pro-Opiomelanocortin