Biphasic insulin secretion in response to a sustained glucose stimulus occurs when rat or human islets are exposed to high levels of the hexose. A transient burst of hormone secretion is followed by a rising and sustained secretory response that, in the perfused rat pancreas, is 25- to 75-fold greater than prestimulatory insulin release rates. This insulin secretory response is paralleled by a significant five- to sixfold increase in the phospholipase C (PLC)-mediated hydrolysis of islet phosphoinositide (PI) pools by high glucose. In contrast, mouse islets, when stimulated under comparable conditions with high glucose, display a second-phase response that is flat and only slightly (two- to threefold) greater than prestimulatory release rates. The minimal second-phase insulin secretory response to high glucose is accompanied by the minimal activation of PLC in mouse islets as well. However, stimulation of mouse islets with the protein kinase C (PKC) activator tetradecanoyl phorbol acetate (TPA) or the muscarinic agonist carbachol, which significantly activates an isozyme of PLC distinct from that activated by high glucose, induces a rising and sustained second-phase insulin secretory response. When previously exposed to high glucose, both rat and human islets respond to subsequent restimulation with an amplified insulin secretory response. They display priming, sensitization, or time-dependent potentiation. In contrast, mouse islets primed under similar conditions with high glucose fail to display this amplified insulin secretory response on restimulation. Mouse islets can, however, be primed by brief exposure to either TPA or carbachol. Finally, whereas rat islets are desensitized by chronic exposure to high glucose, mouse islet insulin secretory responses are relatively immune to this adverse effect of the hexose. These and other findings are discussed in relationship to the role being played by agonist-induced increases in the PLC-mediated hydrolysis of islet phosphoinositide pools and the activation of PKC in these species-specific insulin secretory response patterns.