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      Immunohistochemical and Biochemical Detection of Low Density Lipoprotein Receptors in Cultured Rat Mesangial Cells

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          Abstract

          The expression of low density lipoprotein (LDL) receptors by cultured rat mesangial cells was demonstrated using immunohistochemical and biochemical methods. Using a mouse monoclonal anti-human LDL receptor antibody, the immunofluorescence technique was applied to cultured mesangial cells and showed granular staining. Immunoblotting analysis of the membrane fraction of cultured mesangial cells showed a single band with a protein of approximately 130,000 molecular weight. In addition, [<sup>125</sup>I]LDL binding and the uptake of [<sup>125</sup>I]LDL by cultured mesangial cells were studied. Binding and uptake both reached an equilibrium after about 30 min of incubation. A 50% reduction in [<sup>125</sup>I]LDL (20 μg protein/ml) binding and uptake occurred when unlabelled LDL was added to cultures at 20-40 μg protein/ml. Addition of high density lipoprotein without apoE to the culture medium did not induce competitive inhibition of [<sup>125</sup>I]LDL binding and uptake by mesangial cells. These findings suggest that cultured mesangial cells have the capacity to express an LDL receptor that regulates the cellular uptake of LDL.

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          Author and article information

          Journal
          NEF
          Nephron
          10.1159/issn.1660-8151
          Nephron
          S. Karger AG
          1660-8151
          2235-3186
          1995
          1995
          17 December 2008
          : 69
          : 3
          : 305-310
          Affiliations
          First Department of Medicine, Teikyo University School of Medicine, Tokyo, Japan
          Article
          188475 Nephron 1995;69:305–310
          10.1159/000188475
          7538633
          © 1995 S. Karger AG, Basel

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          Page count
          Pages: 6
          Categories
          Original Paper

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