Quantitative analysis of the cardiac fibroblast transcriptome-implications for NO/cGMP signaling.
Cardiac fibroblasts regulate tissue repair and remodeling in the heart. To quantify transcript levels in these cells we performed a comprehensive gene expression study using serial analysis of gene expression (SAGE). Among 110,169 sequenced tags we could identify 30,507 unique transcripts. A comparison of SAGE data from cardiac fibroblasts with data derived from total mouse heart revealed a number of fibroblast-specific genes. Cardiac fibroblasts expressed a specific collection of collagens, matrix proteins and metalloproteinases, growth factors, and components of signaling pathways. The NO/cGMP signaling pathway was represented by the mRNAs for alpha(1) and beta(1) subunits of guanylyl cyclase, cGMP-dependent protein kinase type I (cGK I), and, interestingly, the G-kinase-anchoring protein GKAP42. The expression of cGK I was verified by RT-PCR and Western blot. To establish a functional role for cGK I in cardiac fibroblasts we studied its effect on cell proliferation. Selective activation of cGK I with a cGMP analog inhibited the proliferation of serum-stimulated cardiac fibroblasts, which express cGK I, but not higher passage fibroblasts, which contain no detectable cGK I. Currently, our data suggest that cGK I mediates the inhibitory effects of the NO/cGMP pathway on cardiac fibroblast growth. Furthermore the SAGE library of transcripts expressed in cardiac fibroblasts provides a basis for future investigations into the pathological regulatory mechanisms underlying cardiac fibrosis.
Smolenski
Albert
A
Institute of Biochemistry II, Medical Faculty, University of Frankfurt, Theodor-Stern-Kai 7, Building 75, D-60590 Frankfurt, Germany. smolenski@biochem2.uni-frankfurt.de
Schultess
Jan
J
Danielewski
Oliver
O
Garcia Arguinzonis
Maisa I
MI
Thalheimer
Petra
P
Kneitz
Susanne
S
Walter
Ulrich
U
Lohmann
Suzanne M
SM
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000378:metabolism; D002455:Cell Division; D006152:Cyclic GMP / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D005355:Fibrosis / Q000473:pathology; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation; D006162:Guanylate Cyclase / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D051379:Mice; D009206:Myocardium / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism*; D020543:Proteome; D012333:RNA, Messenger / Q000378:metabolism; D000074261:Receptors, Enterotoxin; D050705:Receptors, Guanylate Cyclase-Coupled; D018000:Receptors, Peptide / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction
D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; C405330:Gkap1 protein, mouse; D047908:Intracellular Signaling Peptides and Proteins; D020543:Proteome; D012333:RNA, Messenger; D018000:Receptors, Peptide; C015482:protein kinase modulator; D009569:Nitric Oxide; D006162:Guanylate Cyclase; D000074261:Receptors, Enterotoxin; D050705:Receptors, Guanylate Cyclase-Coupled; D006152:Cyclic GMP