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      Regulation of the Expression of Cyclic GMP-Dependent Protein Kinase by Cell Density in Vascular Smooth Muscle Cells

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          Cyclic GMP-dependent protein kinase (cGMP kinase) is the major receptor protein for cGMP in vascular smooth muscle. Vascular smooth muscle cells (VSMC) isolated from the rat aorta express type I cGMP kinase at high levels, but expression decreases markedly upon passage of the cells. In primary or early passage, the expression of cGMP kinase is lowest when cells are plated at low density as assessed by immunological and Northern analyses. Expression increases at confluence and is maintained in postconfluent cultures. With repeated passaging, however, the levels of cGMP kinase decrease even in confluent and postconfluent cultures so that after several passages enzyme levels are undetectable. The decrease in expression in passaged cells is not due to exposure to serum-derived growth factors, but rather on the repeated exposure of cells to conditions in which cell density is reduced (i.e., subculturing). These results indicate that aortic VSMC grown at low density or those repetitively passaged have reduced expression of cGMP kinase, and thus may not represent appropriate cultures with which to investigate the role of nitric oxide and cGMP in VSMC function.

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          Author and article information

          J Vasc Res
          Journal of Vascular Research
          S. Karger AG
          23 September 2008
          : 31
          : 6
          : 330-337
          aDepartment of Pathology, Division of Molecular and Cellular Pathology, University of Alabama at Birmingham, Ala., and bDepartment of Medicine, Division of Hematology/Oncology, Northwestern University Medical School, Chicago, Ill., USA
          159061 J Vasc Res 1994;31:330–337
          © 1994 S. Karger AG, Basel

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          Pages: 8
          Research Paper


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