Endothelin-3 attenuates the cyclic GMP responses to C-type natriuretic peptide in cultured mouse astrocytes

Journal of Neuroscience Research
V T YeungC S Cockram

Abstract

The effect of endothelin-3 (ET-3) on cyclic GMP (cGMP) responses to C-type natriuretic peptide (CNP) was studied in primary cultures of mouse astrocytes. Attenuation of CNP-stimulated cGMP formation by ET-3 was time-dependent, with maximum inhibition achieved at 30 min of preincubation. ET-3 suppressed cGMP production in response to 10 nM CNP in a dose-dependent fashion, with an IC50 of 0.04 nM and a maximal inhibitory concentration of 1 microM, which led to a 66% reduction of the cGMP increment from 45.0 +/- 4.2 pmol/mg protein to 15.4 +/- 2.6 pmol/mg protein. ET-1, ET-2, and ET-3 were equipotent in suppressing the CNP-induced cGMP response, suggesting that this effect was mediated by ETB receptors. Staurosporine, Ro 31-8220, calcium-free medium, nifedipine, verapamil, lanthanum, thapsigargin, BAPTA, W7, calmidazolium, U-73122, neomycin, quinacrine, wortmannin, herbimycin-A, okadaic acid, and sodium orthovanadate failed to block the effect of ET-3. Cycloheximide (100 microM), however, partially but significantly reversed the inhibitory effect of ET-3 on CNP-induced cGMP from 48.2 to 73.3% of the control value. The results support the premise that ET-3 and CNP interact within the central nervous system. The data also suggest th...Continue Reading

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