Hormone Coexpression in the Adult Toad Endocrine Pancreas: Double-Label Immunofluorescence under Basal Conditions and after Glucose Injection

We have investigated the type and frequency of hormone coexpression in the endocrine pancreas of amphibians both under basal conditions and after sustained glucose loading. Adult male specimens of the wild toad Bufo arenarum were injected with a 50% (w/v) glucose solution (2 g/100 g) for 2 days, while control animals received an equal volume of the vehicle. Serum glucose levels were measured at the time of sacrifice and the pancreatic free lobe was processed for light microscopy. A double-labeling immunofluorescence study was performed for the detection of insulin (I), glucagon (G), somatostatin (S), and pancreatic polypeptide (PP). Heterospecific antisera against the following hormone combinations were used for their detection and immunocytochemical localization: I+G, I+PP, G+PP, S+G, and S+PP; visualization of the reacted IgG's was effected by fluorescein- and rhodamine-conjugated species-specific antibodies as fluorophores. Intracellular hormone coexpression was found to occur in the combinations G+PP, S+G, and S+PP. Moreover, glucose administration caused, together with a marked hyperglycemia (123±17 vs 23±1 mg/dl; P