Inositol phospholipids localized to caveolae in rat heart are regulated by alpha1-adrenergic receptors and by ischemia-reperfusion.
Postischemic reperfusion of rat or mouse hearts causes generation of inositol (1,4,5)trisphosphate [Ins(1,4,5)P3] and the initiation of arrhythmias. In the current study we investigated the possibility that the enhanced Ins(1,4,5)P3 generation in postischemic reperfusion was associated with an increased availability of the precursor lipid phosphatidylinositol(4,5)bisphosphate (PIP2) for alpha1-adrenergic receptor-activated phospholipase C (PLC). Isolated-perfused rat hearts were labeled with [3H]inositol and subjected to ischemia-reperfusion or stimulation with norepinephrine under normoxic conditions. Caveolar fractions were prepared by buoyant density sucrose gradient centrifugation. [3H]PIP2 was concentrated in caveolae, along with Galphaq and PLCbeta1b. Caveolae contained only 27.3 +/- 6.9% (means +/- SE, n = 6) of the total alpha1-adrenergic receptor complement of the heart. These did not migrate to PIP2-containing caveolar fractions with norepinephrine stimulation under normoxic conditions, even though caveolar PIP2 was depleted. In contrast, [3H]PIP2 in caveolae increased during 2 min of reperfusion, independently of norepinephrine release and thus of alpha1-adrenergic receptor activation. The increased PIP2 in the caveolar fractions where signaling proteins are concentrated may be critical for the heightened generation of Ins(1,4,5)P3 in early reperfusion.
Lanzafame
Alfred A
AA
Cellular Biochemistry Laboratory, Baker Heart Research Institute, Melbourne, Victoria, Australia.
Turnbull
Lynne
L
Amiramahdi
Fatemeh
F
Arthur
Jane F
JF
Huynh
Huy
H
Woodcock
Elizabeth A
EA
D000818:Animals; D021941:Caveolae / Q000378:metabolism*; D066298:In Vitro Techniques; D008297:Male; D021962:Membrane Microdomains / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D010716:Phosphatidylinositols / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018340:Receptors, Adrenergic, alpha-1 / Q000378:metabolism*; D015427:Reperfusion Injury / Q000378:metabolism*
D010716:Phosphatidylinositols; D018340:Receptors, Adrenergic, alpha-1