Exogenous nitric oxide donation causes desensitization of arteriolar relaxing activity in vivo: an intravital analysis in mice.

ABSTRACT

BACKGROUND: The NO/cGMP pathway plays a crucial role in regulation of tissue perfusion. However, a NO-induced desensitization of cGMP-mediated relaxation has been reported in isolated tissue. To examine whether a similar phenomenon can be detected in vivo, we analyzed relaxations of microvessels in response to repeated applications of NO. MATERIALS AND METHODS: The investigations were performed by means of dynamic intravital fluorescence microscopy in the dorsal skinfold chamber of female balb/mice. First, the microvasculature was maximally preconstricted by the application of the vasoconstrictor 5-hydroxytryptamine. Subsequently, relaxation was induced by applying an NO-donator, the S-nitrosoglutathione, to the contracted vessels. Following buffer exchange, constriction and relaxation were repeated. Drugs were given topically into the chamber, directly onto the skin muscle. The response of arterioles to topical administration of vasoactive drugs was determined as the change of the diameter, and quantified using standard software. RESULTS: The relaxation of arterioles was reduced after repetitive application. The short pretreatment with NO-donor entailed a reduced relaxation of arterioles in response to following application. The absolute change in vessel diameter induced by S-nitrosoglutathione was significantly reduced from 21 µm to 16 µm after the first and the second application, respectively. However, the data also revealed a noticeable reduction of the constricting activity of 5-hydroxytryptamine during the second application, indicating a possible desensitization of the 5-hydroxytryptamine response or a humoral and/or neuronal compensatory mechanisms. CONCLUSIONS: The NO-induced cGMP-mediated relaxation of microvessels was quantified, and the phenomenon of desensitization visualized in vivo by means of dynamic fluorescence microscopy.