Quisqualate and carbachol-induced increases in intrasynaptosomal free calcium are mediated by different products of phospholipid hydrolysis.
The mechanisms by which quisqualate and carbachol increase intrasynaptosomal free calcium ([Ca2+]i) were studied in rat cortical synaptosomes. Quisqualate (0.01-100 microM) and carbachol (100-1000 microM) increased [Ca2+]i in Fura-2 acetoxymethyl ester (Fura-2 AM)-loaded synaptosomes. The resting level of [Ca2+]i was 118 nM. The maximum increase (55%) was produced by 10 microM quisqualate which had an EC50 of 0.2 microM. The maximum increase (28%) elicited by carbachol occurred at 1000 microM and the EC50 was 30 microM. The stimulatory effects of quisqualate on [Ca2+]i were blocked by heparin (100 I.U.) but not by staurosporine (1 microM), nifedipine (1 microM) or omega-conotoxin fraction GVIA (omega-CgTx) (0.5 microM). On the other hand, the effects of carbachol on [Ca2+]i were abolished by staurosporine, nifedipine or omega-CgTx but not by heparin. Carbachol (100 microM) also significantly increased 45Ca accumulation into either resting or K+ (30 mM)-depolarised synaptosomes and these effects were inhibited by staurosporine and nifedipine. Quisqualate (10 microM) had no effect on 45Ca accumulation under resting or depolarised conditions. When quisqualate and carbachol were used in combination, there were apparently additive effects on [Ca2+]i but not on 45Ca accumulation. It is concluded that carbachol increases [Ca2+]i by facilitating Ca2+ entry through L-type Ca2+ channels via a 1,2-diacylglycerol (DAG)-protein kinase C (PKC)-dependent pathway while quisqualate mobilizes Ca2+ from inositol 1,4,5-trisphosphate (IP3)-sensitive stores.
Xiang
J Z
JZ
Department of Neuroscience, Institute of Psychiatry, London, U.K.
Brammer
M J
MJ
Campbell
I C
IC
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002132:Calcium Radioisotopes; D002217:Carbachol / Q000494:pharmacology*; D066298:In Vitro Techniques; D008297:Male; D010743:Phospholipids / Q000378:metabolism*; D016318:Quisqualic Acid / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism*
D002121:Calcium Channel Blockers; D002132:Calcium Radioisotopes; D010743:Phospholipids; D016318:Quisqualic Acid; D002217:Carbachol; D002118:Calcium