Regulation of vascular smooth muscle soluble guanylate cyclase activity, mRNA, and protein levels by cAMP-elevating agents

Andreas Papapetropoulos, Nandor Marczin, Gloria Mora, Antonio Milici, Ferid Murad, John D. Catravas

Research output: Contribution to journalArticle

79 Citations (Scopus)

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 100 μmol/L isobutylmethylxanthine o 0.01 to 10 μmol/L forskolin led to a time- and concentration-dependent decease 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-release 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 α1 - and β1-subunits of sGC were decreased in cells pretreated with isobutylmethylxanthine and forskolin but not with dideoxyforskolin (inactive analogue). Moreover, protein levels for the sGC α1-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.

Original languageEnglish (US)
Pages (from-to)696-704
Number of pages9
JournalHypertension
Volume26
Issue number4
DOIs
StatePublished - Jan 1 1995
Externally publishedYes

Fingerprint

Vascular Smooth Muscle
Colforsin
Messenger RNA
Nitroprusside
Proteins
Gene Expression Regulation
Second Messenger Systems
Cyclic AMP-Dependent Protein Kinases
Reverse Transcriptase Polymerase Chain Reaction
Smooth Muscle Myocytes
Blood Vessels
Soluble Guanylyl Cyclase
Down-Regulation
Western Blotting
High Pressure Liquid Chromatography
Gene Expression

Keywords

  • cyclic AMP
  • cyclic GMP
  • guanylate cyclase
  • nitroprusside

ASJC Scopus subject areas

  • Internal Medicine

Cite this

Regulation of vascular smooth muscle soluble guanylate cyclase activity, mRNA, and protein levels by cAMP-elevating agents. / Papapetropoulos, Andreas; Marczin, Nandor; Mora, Gloria; Milici, Antonio; Murad, Ferid; Catravas, John D.

In: Hypertension, Vol. 26, No. 4, 01.01.1995, p. 696-704.

Research output: Contribution to journalArticle

Papapetropoulos, Andreas ; Marczin, Nandor ; Mora, Gloria ; Milici, Antonio ; Murad, Ferid ; Catravas, John D. / Regulation of vascular smooth muscle soluble guanylate cyclase activity, mRNA, and protein levels by cAMP-elevating agents. In: Hypertension. 1995 ; Vol. 26, No. 4. pp. 696-704.
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