The NO signaling pathway differentially regulates KCC3a and KCC3b mRNA expression.
Nitric oxide (NO) donors and protein kinase G (PKG) acutely up-regulate K-Cl cotransporter-1 and -3 (KCC1 and KCC3) mRNA expression in vascular smooth muscle cells (VSMCs). Here, we report the presence, relative abundance, and regulation by sodium nitroprusside (SNP) of the novel KCC3a and KCC3b mRNAs, in primary cultures of rat VSMCs. KCC3a and KCC3b mRNAs were expressed in an approximate 3:1 ratio, as determined by semiquantitative RT-PCR analysis. SNP as well as YC-1 and 8-Br-cGMP, a NO-independent stimulator of soluble guanylyl cyclase (sGC) and PKG, respectively, increased KCC3a and KCC3b mRNA expression by 2.5-fold and 8.1-fold in a time-dependent manner, following a differential kinetics. Stimulation of the NO/sGC/PKG signaling pathway with either SNP, YC-1, or 8-Br-cGMP decreased the KCC3a/KCC3b ratio from 3.0+/-0.4 to 0.9+/-0.1. This is the first report on a differential regulation by the NO/sGC/PKG signaling pathway of a cotransporter and of KCC3a and KCC3b mRNA expression.
Di Fulvio
Mauricio
M
Department of Pharmacology and Toxicology, Wright State University, School of Medicine, 3640 Colonel Glenn Highway, Biological Sciences Building, Room 152-6, Dayton, OH 45435-0002, USA.
Lauf
Peter K
PK
Adragna
Norma C
NC
D000818:Animals; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000494:pharmacology; D017869:Cyclic GMP-Dependent Protein Kinases / Q000502:physiology; D005786:Gene Expression Regulation; D006162:Guanylate Cyclase; D007191:Indazoles / Q000494:pharmacology; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism* / Q000502:physiology; D009599:Nitroprusside / Q000494:pharmacology; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000502:physiology; D015398:Signal Transduction; D000071756:Soluble Guanylyl Cyclase; D027981:Symporters / Q000096:biosynthesis* / Q000235:genetics; D013873:Thionucleotides / Q000494:pharmacology; D000096922:K Cl- Cotransporters
D006152:Cyclic GMP; D017869:Cyclic GMP-Dependent Protein Kinases; D006162:Guanylate Cyclase; D007191:Indazoles; D009569:Nitric Oxide; D009599:Nitroprusside; D020033:Protein Isoforms; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; D000071756:Soluble Guanylyl Cyclase; D027981:Symporters; D013873:Thionucleotides; C492281:Slc12a6 protein, rat; D000096922:K Cl- Cotransporters; C083763:8-bromoguanosino-3',5'-cyclic monophosphorothioate; C090937:3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole