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Am J Physiol Heart Circ Physiol 278: H1421-H1428, 2000;
0363-6135/00 $5.00
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Vol. 278, Issue 5, H1421-H1428, May 2000

Sphingomyelinase and ceramide analogs induce contraction and rises in [Ca2+]i in canine cerebral vascular muscle

Tao Zheng1, Wenyan Li1, Jun Wang2, Bella T. Altura1,3,4, and Burton M. Altura1,3,4

Departments of 1 Physiology, 2 Anesthesiology, and 3 Medicine and 4 Center for Cardiovascular and Muscle Research, State University of New York, Health Science Center at Brooklyn, Brooklyn, New York 11203

Studies were designed to investigate effects of neutral sphingomyelinase (N-SMase) and ceramide analogs as well as phosphorylcholine on vascular tone and Ca2+ mobilization in isolated canine cerebral arterial smooth muscle. N-SMase (0.001-0.1 U/ml) provoked a gradual but sustained vasoconstriction of arterial rings in a concentration-related manner that was endothelium independent. Incubation of denuded arterial rings in Ca2+-free medium or pretreatment with verapamil in extracellular Ca2+ resulted in a reduction of the N-SMase-evoked constriction. Exposure of arterial rings to 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA)-AM did not, however, result in a reduction of N-SMase-induced constriction. Both staurosporine and bisindolymaleimide I attenuated N-SMase-induced contractions to 66% and 72% of control, respectively. N-SMase caused gradual and sustained rises in intracellular Ca2+ concentration ([Ca2+]i) in primary cultured cerebral vascular smooth muscle cells. Pretreatment of these cultured cells with nimodipine and verapamil caused a steady decline in N-SMase-induced rises in [Ca2+]i. Exposure of the cells to Ca2+-free solution reversed the [Ca2+]i-induced rise triggered by N-SMase to the resting baseline. Both C8 and C16 ceramide (10-9-10-6 M), but not phosphorylcholine, constricted denuded canine arterial rings in a concentration-related manner and elevated [Ca2+]i. Our results suggest that the sphingomyelin-signaling pathway, via a probable release of ceramide molecules, may play an important role in regulation of cerebral arterial wall tone.

intracellular calcium; sphingolipids; signal transduction; smooth muscle contraction; cerebral vascular tone


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