Triggering of insulin release by a combination of cAMP signal and nutrients: an ATP-sensitive K+ channel-independent phenomenon.
Nutrient augmentation of Ca(2+)-triggered insulin release occurs in an ATP-sensitive K(+) (K(ATP)) channel--independent manner. Here, using rat islets, we explored the possibility of the K(ATP) channel-independent nutrient triggering of insulin release. In the presence of 250 micromol/l diazoxide, simultaneous application of forskolin and 16.7 mmol/l glucose strongly stimulated insulin release: fourfold and eightfold increases with 1 and 30 micromol/l forskolin, respectively. alpha-Ketoisocaproate (KIC) and 3-isobutylmethylxanthine (IBMX) could be used in place of glucose and forskolin, respectively, to trigger insulin release in the presence of diazoxide. Triggering of insulin release by a combination of nutrients and forskolin was not attenuated by 10 micromol/l nifedipine (a blocker of voltage-dependent Ca(2+) channels) and 2 micromol/l thapsigargin (an inhibitor of intracellular Ca(2+)-ATPase), ascertaining independence of this phenomenon from Ca(2+) entry and from intracellular Ca(2+) liberation. As anticipated, the action of glucose and KIC was greatly (>80%) suppressed by inhibition of mitochondrial metabolism by 2 mmol/l sodium azide (NaN(3)). A combination of palmitate and dimethyl glutamate (a cell-permeable glutamate donor), but not either one alone, weakly but unequivocally triggered insulin release when applied simultaneously with forskolin. In this case, however, mitochondrial poisoning by azide was without effect. The finding suggests that a combination of induced palmitoylation and cytosolic glutamate accumulation partially reconstituted signaling beyond mitochondrial metabolism in the beta-cell upon glucose stimulation. In conclusion, a combination of cAMP signal and nutrients potently triggers insulin release under full activation of the K(ATP) channel, indicating the multiplicity of driving force for insulin exocytosis.
Komatsu
Mitsuhisa
M
Department of Aging Medicine and Geriatrics, Shinshu University School of Medicine, Matsumoto, Japan. mitsuk@hsp.md.shinshu-u.ac.jp
Sato
Yoshihiko
Y
Yamada
Satoko
S
Yamauchi
Keishi
K
Hashizume
Kiyoshi
K
Aizawa
Toru
T
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D003981:Diazoxide / Q000494:pharmacology; D005934:Glucagon / Q000494:pharmacology; D052216:Glucagon-Like Peptide 1; D005947:Glucose / Q000494:pharmacology*; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism*; D008297:Male; D010446:Peptide Fragments / Q000494:pharmacology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D015221:Potassium Channels / Q000378:metabolism*; D011498:Protein Precursors / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D019810:Sodium Azide / Q000494:pharmacology; D014665:Vasodilator Agents / Q000494:pharmacology
D007328:Insulin; D010446:Peptide Fragments; D010726:Phosphodiesterase Inhibitors; D015221:Potassium Channels; D011498:Protein Precursors; D014665:Vasodilator Agents; D005576:Colforsin; D052216:Glucagon-Like Peptide 1; D000255:Adenosine Triphosphate; D005934:Glucagon; D019810:Sodium Azide; D000242:Cyclic AMP; D005947:Glucose; D003981:Diazoxide; D002118:Calcium; D015056:1-Methyl-3-isobutylxanthine