Regulation of vascular smooth muscle soluble guanylate cyclase activity, mRNA, and protein levels by cAMP-elevating agents
| Autor: | Gloria Mora, Andreas Papapetropoulos, John D. Catravas, Ferid Murad, Antonio Milici, Nandor Marczin |
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| Rok vydání: | 1995 |
| Předmět: |
Vascular smooth muscle
Transcription Genetic Immunoblotting Molecular Sequence Data Biology Polymerase Chain Reaction Muscle Smooth Vascular Rats Sprague-Dawley Transcription (biology) Internal Medicine medicine Cyclic AMP Animals RNA Messenger Cyclic GMP Cells Cultured Chromatography High Pressure Liquid chemistry.chemical_classification Regulation of gene expression Messenger RNA Base Sequence Guanylate cyclase activity RNA Intracellular Membranes Rats medicine.anatomical_structure Enzyme chemistry Biochemistry Solubility Guanylate Cyclase Molecular Probes Blood vessel |
| Zdroj: | Scopus-Elsevier |
| ISSN: | 0194-911X |
| Popis: | Abstract Although the biochemical properties of soluble guanylate cyclase (sGC) have been extensively studied, little is known about the regulation of gene expression of sGC subunits by second messengers. cAMP analogues and elevating agents have been previously shown to alter gene expression in vascular cells. The aim of the present study was to investigate the effects of cAMP-elevating agents on sodium nitroprusside–stimulated sGC activity and to correlate activity changes with mRNA and protein levels in cultured rat aortic smooth muscle cells. Pretreatment of cells with 50 to 1000 μmol/L isobutylmethylxanthine or 0.01 to 10 μmol/L forskolin led to a time- and concentration-dependent decrease in sodium nitroprusside–induced cGMP accumulation, first evident after 3 hours of pretreatment with forskolin and 6 hours of pretreatment with isobutylmethylxanthine. Incubation of cells with a protein kinase A–selective inhibitor (H89 or KT 5720) partially or fully prevented the downregulation in sodium nitroprusside–induced cGMP accumulation caused by cAMP-elevating agents. Quantification of reverse transcriptase–polymerase chain reaction products by high-performance liquid chromatography revealed that mRNA for both α 1 - and β 1 -subunits of sGC were decreased in cells pretreated with isobutylmethylxanthine and forskolin but not with dideoxyforskolin (inactive analogue). Moreover, protein levels for the sGC α 1 -subunit of cells pretreated with isobutylmethylxanthine and forskolin but not with dideoxyforskolin were decreased as indicated by Western blot analysis. These data indicate that cAMP-elevating agents decrease sGC activity, possibly by decreasing mRNA or protein levels or both. |
| Databáze: | OpenAIRE |
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