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Am J Physiol Heart Circ Physiol 280: H1222-H1231, 2001;
0363-6135/01 $5.00
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Vol. 280, Issue 3, H1222-H1231, March 2001

In vivo assessment of microvascular nitric oxide production and its relation with blood flow

X. F. Figueroa, A. D. Martínez, D. R. González, P. I. Jara, S. Ayala, and M. P. Boric

Unidad de Regulación Neurohumoral, Departamento de Ciencias Fisiológicas, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile 6513492

To assess the hypothesis that microvascular nitric oxide (NO) is critical to maintain blood flow and solute exchange, we quantified NO production in the hamster cheek pouch in vivo, correlating it with vascular dynamics. Hamsters (100-120 g) were anesthetized and prepared for measurement of microvessel diameters by intravital microscopy, of plasma flow by isotopic sodium clearance, and of NO production by chemiluminescence. Analysis of endothelial NO synthase (eNOS) location by immunocytochemistry and subcellular fractionation revealed that eNOS was present in arterioles and venules and was 67 ± 7% membrane bound. Basal NO release was 60.1 ± 5.1 pM/min (n = 35), and plasma flow was 2.95 ± 0.27 µl/min (n = 29). Local NO synthase inhibition with 30 µM Nomega -nitro-L-arginine reduced NO production to 8.6 ± 2.6 pmol/min (-83 ± 5%, n = 9) and plasma flow to 1.95 ± 0.15 µl/min (-28 ± 12%, n = 17) within 30-45 min, in parallel with constriction of arterioles (9-14%) and venules (19-25%). The effects of Nomega -nitro-L-arginine (10-30 µM) were proportional to basal microvascular conductance (r = 0.7, P < 0.05) and fully prevented by 1 mM L-arginine. We conclude that in this tissue, NO production contributes to 35-50% of resting microvascular conductance and plasma-tissue exchange.

nitric oxide chemiluminescence; subcellular endothelial nitric oxide synthase distribution; Nomega -nitro-L-arginine; vascular conductance; hamster cheek pouch


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