title: Nitric oxide signaling pathway regulates potassium chloride cotransporter-1 mRNA expression in vascular smooth muscle cells.
abstract: Rat vascular smooth muscle cells (VSMCs) express at least two mRNAs for K-Cl cotransporters (KCC): KCC1 and KCC3. cGMP-dependent protein kinase I regulates KCC3 mRNA expression in these cells. Here, we show evidence implicating the nitric oxide (NO)/cGMP signaling pathway in the expression of KCC1 mRNA, considered to be the major cell volume regulator. VSMCs, expressing soluble guanylyl cyclase (sGC) and PKG-I isoforms showed a time- and concentration-dependent increase in KCC1 mRNA levels after treatment with sodium nitroprusside as demonstrated by semiquantitative RT-PCR. sGC-dependent regulation of KCC1 mRNA expression was confirmed using YC-1, a NO-independent sGC stimulator. The sGC inhibitor LY83583 blocked the effects of sodium nitroprusside and YC-1. Moreover, 8-Br-cGMP increased KCC1 mRNA expression in a concentration- and time-dependent fashion. The 8-Br-cGMP effect was partially blocked by KT5823 but not by actinomycin D. However, actinomycin D and cycloheximide increased basal KCC1 mRNA in an additive manner, suggesting different mechanisms of action for both drugs. These findings suggest that in VSMCs, the NO/cGMP-signaling pathway participates in KCC1 mRNA regulation at the post-transcriptional level.
authors.affiliation: Department of Pharmacology, School of Medicine, Wright State University, Dayton, Ohio 45435-0002, USA.
authors.forename: M
authors.initials: M
authors.lastname: Di Fulvio
authors.forename: P K
authors.initials: PK
authors.lastname: Lauf
authors.forename: N C
authors.initials: NC
authors.lastname: Adragna
mesh_terms: D000470:Alkaloids / Q000494:pharmacology; D000634:Aminoquinolines / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport; D002227:Carbazoles; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007211:Indoles; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D009569:Nitric Oxide / Q000378:metabolism* / Q000494:pharmacology; D009599:Nitroprusside / Q000494:pharmacology; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D027981:Symporters / Q000096:biosynthesis*; D013997:Time Factors; D000096922:K Cl- Cotransporters
chemical_list: D000470:Alkaloids; D000634:Aminoquinolines; D002227:Carbazoles; D006152:Cyclic GMP; D003513:Cycloheximide; D003609:Dactinomycin; D004791:Enzyme Inhibitors; D007211:Indoles; D009569:Nitric Oxide; D009599:Nitroprusside; D012333:RNA, Messenger; D027981:Symporters; D000096922:K Cl- Cotransporters; C073601:KT 5823; C016276:8-bromocyclic GMP; C041715:6-anilino-5,8-quinolinedione