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Am J Physiol Heart Circ Physiol (October 7, 2005). doi:10.1152/ajpheart.00052.2005
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Submitted on January 18, 2005
Accepted on October 1, 2005

SUPEROXIDE ANION IS ELEVATED IN SYMPATHETIC NEURONS IN DOCA-SALT HYPERTENSION VIA ACTIVATION OF NADPH OXIDASE

Xiaoling Dai1, Xian Cao1, and David L Kreulen1*

1 Physiology, Michigan State University, East Lansing, MI, USA; Neuroscience Program, Michigan State University, East Lansing, MI, USA

* To whom correspondence should be addressed. E-mail: dkreulen{at}msu.edu.

Superoxide anion (O2.-) production is elevated in sympathetic ganglion neurons and in the vasculature of hypertensive animals but it is not known what enzymatic pathway(s) are responsible for O2.- production. In order to determine the pathway(s) of O2.- production in sympathetic neurons, we examined the presence of mRNA of NADPH oxidase subunits in sympathetic ganglionic neurons and differentiated PC-12 cells. The mRNAs for NADPH oxidase subunits p47phox, p22phox, gp91phox and NOX1 were present in sympathetic neurons and PC-12 cells whereas the NOX4 homologue was present in sympathetic neurons but not PC-12 cells. Freshly dissociated celiac ganglion neurons from normal rats and PC-12 cells produced O2.- when treated with the PKC activator phorbol myristate acetate (PMA), O2.- production increased by 317% and 254%, respectively. The PMA-evoked increases were reduced by pretreatment with the NADPH oxidase inhibitor apocynin. These findings indicate that NADPH oxidase is the primary source of O2.- in sympathetic ganglion neurons. When celiac ganglia from hypertensive rats were incubated with apocynin, O2.- levels were reduced to the same levels as normotensive animals indicating that NADPH oxidase activity accounted for the elevated O2.- levels in hypertensive animals. To test this latter finding we compared NADPH oxidase activity in extracts of prevertebral sympathetic ganglia of DOCA-salt hypertensive rats and Sham-operated rats. NADPH oxidase activities were 49.9% and 78.6% higher in sympathetic ganglia of DOCA rats compared to normotensive controls when using {beta}-NADH and {beta}-NADPH as substrates, respectively. Thus, elevated O2.- levels in hypertension may be a result of the increased activity of NADPH oxidase in postganglionic sympathetic neurons.




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