Nutrient augmentation of Ca2+-dependent and Ca2+-independent pathways in stimulus-coupling to insulin secretion can be distinguished by their guanosine triphosphate requirements: studies on rat pancreatic islets.
To delineate the underlying mechanisms by which glucose augments both Ca2+-dependent and Ca2+-independent insulin release, the latter induced by the simultaneous activation of protein kinases A and C, we examined the effects of GTP depletion by mycophenolic acid (MPA), an inhibitor of GTP synthesis, on the augmentation of insulin release from rat pancreatic islets. MPA treatment reduced GTP content by 30-40% and completely abolished glucose-induced augmentation of Ca2+-independent insulin release. Thus, this pathway is extremely sensitive to a decrease in cellular GTP content. Complete inhibition was also observed in islets treated with MPA plus adenine, to maintain ATP levels, under which conditions GTP is selectively depleted. Provision of guanine, which increases the activity of a salvage pathway for GTP synthesis and normalizes GTP content, completely reversed the inhibitory effect of MPA. Neither glucose utilization nor glucose oxidation was affected by MPA. The augmentation of Ca2+-independent insulin release by several other metabolizable nutrients including alpha-ketoisocaproic acid (KIC) was also inhibited by MPA. In sharp contrast, augmentation of Ca2+-dependent insulin release by KIC was resistant to GTP depletion, indicating that nutrient-induced augmentation of the Ca2+-dependent- and Ca2+-independent secretory pathways can be differentiated by GTP dependency. We interpret these data in accord with current knowledge concerning the two known stimuli for exocytosis, Ca2+ and GTP (independently of Ca2+). We propose that both Ca2+-dependent and Ca2+-independent augmentation occurs via one metabolic pathway acting upon Ca2+- and upon GTP-stimulated exocytosis. Activation of PKA and PKC stimulates the GTP-sensitive exocytosis.
Komatsu
M
M
Department of Pharmacology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853-6401, USA.
Noda
M
M
Sharp
G W
GW
D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D002118:Calcium / Q000502:physiology*; D005576:Colforsin / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000032:analysis / Q000502:physiology*; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000378:metabolism*; D008297:Male; D009173:Mycophenolic Acid / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D007328:Insulin; D005576:Colforsin; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D009173:Mycophenolic Acid; D005947:Glucose; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium