title: Inhibition of adrenal cell aldosterone synthesis by endogenous nitric oxide release.
abstract: Adrenal zona glomerulosa (ZG) cells do not contain nitric oxide (NO) synthase (NOS). We conferred endothelial NOS activity onto adrenal ZG cells through transduction with a recombinant adenovirus encoding the endothelial NOS gene (AdeNOS) to determine the effect of endogenous NO on aldosterone synthesis. A 135-kDa protein band immunoreactive to anti-endothelial NOS antibody was observed in Western blots of AdeNOS-transduced ZG cells but not in control cells or cells transduced with adenovirus encoding the beta-galactosidase gene (AdbetaGal). Nitrate/nitrite production in AdeNOS-transduced ZG cells increased from 0.15+/-0.01 to 0.27+/-0.01 micromol/L after stimulation with 1 nmol/L angiotensin II. The treatment of AdeNOS-transduced cells with 30 micromol/L L-nitro-arginine decreased angiotensin II-stimulated nitrite production from 0.27+/-0. 01 to 0.17+/-0.01 micromol/L. Basal and angiotensin II-stimulated nitrite production was not increased in AdbetaGal-transduced or control cells. AdeNOS-transduced cells demonstrated diaminofluorescein-2 diacetate fluorescence, which was blocked by pretreatment with L-nitro-arginine. Angiotensin II-stimulated aldosterone synthesis decreased from 5123+/-177 pg/mL in AdbetaGal-transduced ZG cells to 72+/-27 pg/mL in AdeNOS-transduced cells. Treatment with the NOS inhibitor thiocitrulline (30 micromol/L) increased angiotensin II-stimulated aldosterone synthesis to 2158+/-45 pg/mL after AdeNOS transduction. These data demonstrate that adenovirus-mediated gene transfer of eNOS in ZG cells results in the expression of active endothelial NOS enzyme and that this endogenous NO production by ZG cells decreases aldosterone synthesis.
authors.affiliation: Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee 53226, USA.
authors.forename: C J
authors.initials: CJ
authors.lastname: Hanke
authors.forename: T
authors.initials: T
authors.lastname: O'Brien
authors.forename: K A
authors.initials: KA
authors.lastname: Pritchard
authors.forename: W B
authors.initials: WB
authors.lastname: Campbell
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000257:Adenoviridae Infections / Q000378:metabolism; D000450:Aldosterone / Q000096:biosynthesis*; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002478:Cells, Cultured; D005452:Fluoresceins; D015971:Gene Expression Regulation, Enzymologic; D018014:Gene Transfer Techniques; D009566:Nitrates / Q000032:analysis / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III; D009573:Nitrites / Q000032:analysis / Q000378:metabolism; D015384:Zona Glomerulosa / Q000166:cytology* / Q000201:enzymology* / Q000821:virology
chemical_list: C112456:4,5-diamino-2-(3,6-dihydroxy-9H-xanthen-9-yl)benzoic acid; D005452:Fluoresceins; D009566:Nitrates; D009573:Nitrites; D009569:Nitric Oxide; D000450:Aldosterone; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III