title: NONOates regulate KCl cotransporter-1 and -3 mRNA expression in vascular smooth muscle cells.
abstract: Nitric oxide (NO) donors regulate KCl cotransport (KCC) activity and cotransporter-1 and -3 (KCC1 and KCC3) mRNA expression in sheep erythrocytes and in primary cultures of rat vascular smooth muscle cells (VSMCs), respectively. In this study, we used NONOates as rapid and slow NO releasers to provide direct evidence implicating NO as a regulator of KCC3 gene expression at the mRNA level. In addition, we used the expression of KCC3 mRNA to further investigate the mechanism of action of these NO donors at the cellular level. Treatment of VSMCs with rapid NO releasers, like NOC-5 and NOC-9, as well as with the direct NO-independent soluble guanylyl cyclase (sGC) stimulator YC-1, acutely increased KCC3 mRNA expression in a concentration- and time-dependent manner. The slow NO releaser NOC-18 had no effect on KCC3 gene expression. A specific NO scavenger completely prevented the NONOate-induced KCC3 mRNA expression. Inhibition of sGC with LY-83583 blocked the NONOate- and YC-1-induced KCC3 mRNA expression. This study shows that in primary cultures of rat VSMCs, the fast NO releasers NOC-9 and NOC-5, but not the slow NO releaser NOC-18, acutely upregulate KCC3 mRNA expression in a NO/sGC-dependent manner.
authors.affiliation: Department of Pharmacology and Toxicology, School of Medicine, Wright State University, 3640 Colonel Glenn Highway, Dayton, Ohio 45435-0002, USA.
authors.forename: Mauricio
authors.initials: M
authors.lastname: Di Fulvio
authors.forename: Peter K
authors.initials: PK
authors.lastname: Lauf
authors.forename: Shalin
authors.initials: S
authors.lastname: Shah
authors.forename: Norma C
authors.initials: NC
authors.lastname: Adragna
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D015870:Gene Expression / Q000187:drug effects; D006162:Guanylate Cyclase; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000502:physiology*; D009569:Nitric Oxide / Q000378:metabolism; D020030:Nitric Oxide Donors / Q000494:pharmacology*; D009603:Nitroso Compounds / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D000071756:Soluble Guanylyl Cyclase; D027981:Symporters / Q000235:genetics*; D014226:Triazenes / Q000494:pharmacology*; D000096922:K Cl- Cotransporters
chemical_list: D006162:Guanylate Cyclase; D009569:Nitric Oxide; D020030:Nitric Oxide Donors; D009603:Nitroso Compounds; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; D000071756:Soluble Guanylyl Cyclase; D027981:Symporters; D014226:Triazenes; D000096922:K Cl- Cotransporters; C114535:1-hydroxy-2-oxo-3-(3-aminopropyl)-3-isopropyl-1-triazene; C093285:NOC 18; C492281:Slc12a6 protein, rat; C099612:NOC 9