Differential distribution of nitric oxide synthase isoforms in the rostral ventrolateral medulla of the rat

We evaluated the distribution of nitric oxide synthase (NOS) isoforms in the rostral ventrolateral medulla (RVLM), the medullary origin of sympathetic neurogenic vasomotor tone, and the contribution of NOS III to the cardiovascular actions of endogenous NO in the RVLM. Adult Sprague-Dawley rats were used. Reverse transcription-polymerase chain reaction or Western blot analysis revealed that NOS I, II or III was expressed in the ventrolateral medulla at the mRNA or protein level under basal conditions. However, laser scanning confocal microscopic analysis of double-immunofluorescence images showed that whereas NOS I or II immunoreactivity colocalized with cells within the confines of the RVLM that stained positively with the neuronal marker, NeuN, NOS III immunoreactivity was associated primarily with blood vessels. Furthermore, bilateral microinjection into the RVLM of the selective NOS III inhibitor, N(5)-(1-iminoethyl)-L-ornithine, elicited minimal alterations in baseline systemic arterial pressure, heart rate or sympathetic vasomotor outflow in rats anesthetized with propofol. We conclude that whereas NOS I and II are present in neurons within the confines of the RVLM, NOS III is associated primarily with blood vessels. Our results further indicate that NOS III does not appear to contribute to the maintenance of basal sympathetic vasomotor outflows and arterial pressure by the endogenous NO at the RVLM.