In rat pancreatic islets, gap junctional subunits (GJS) occur under two different configurations, namely in linear single strands and in polygonal particle aggregates. The present freeze-fracture study demonstrates that GJS can rapidly (dis)assemble into one of these membrane specializations without changes in their total number. Isolation of the pancreatic gland and its perfusion at 2.8 mM glucose is accompanied by a decrease in polygonally packed GJS from 46 to 16%. A rise in medium glucose concentration is followed, within 10 min, by a dose-dependent increase in the percent polygonal particles. This glucose effect on gap junction configuration is calcium dependent and reversible upon glucose removal; it is still entirely detectable when protein synthesis is blocked by cycloheximide. These results indicate that islet gap junctions are dynamic structures that rapidly adjust their configuration to extracellular regulators of beta-cell function. In the light of previous observations, it is suggested that this rapid (dis)assembly of gap junctional structures be considered as a component in the ionic and metabolic coupling between insulin-containing beta-cells of the pancreas.