Immunohistochemical localization of cellubrevin on secretory granules in pancreatic B-cells

Abstract

Cellubrevin is one of the proteins involved in the docking and fusion of secretory granules to the plasma membrane. It has been reported that cellubrevin is widely distributed in both neural and non-neural cells, including insulin-secreting B-cells. This study aims to demonstrate by immunohistochemical techniques that cellubrevin is localized in insulin- secreting cells and further to examine whether it might occur in glucagon- and somatostatin-secreting cells in the pancreatic islet in the rat and mouse. We used the polyclonal antibody against the N-terminal peptide whose specificity was confirmed by Western blot analysis. Double immunostaining demonstrated that cellubrevin was localized in insulin-containing cells, but both glucagon-containing and somatostatin-containing cells lacked the immunoreactivity. Immuno-electron microscopic analysis revealed the localization of cellubrevin on the margin of secretory granules near the plasma membrane but not in the granules closer to the nucleus. These observations support the view that cellubrevin in the pacreatic islet is expressed on the membrane of the secretory granules in B-cells at the stage of exocytosis.