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Journal of Histochemistry and Cytochemistry, Vol. 46, 291-300, Copyright © 1998, The Histochemical Society, Inc.

ARTICLE

Binding of Injected Laminin to Developing Kidney Glomerular Mesangial Matrices and Basement Membranes In Vivo

Ruixue Wanga, Danika Moorer–Hickmana, Patricia L. St. Johna, and Dale R. Abrahamsona
a Department of Cell Biology, University of Alabama at Birmingham, Birmingham, Alabama

Correspondence to: Dale R. Abrahamson, Dept. of Cell Biology, U. of Alabama at Birmingham, 6th Floor Volker Hall, 1670 University Blvd, Birmingham, AL 35294-0019.

During glomerular development, subendothelial and -epithelial basement membrane layers fuse to produce the glomerular basement membrane (GBM) shared by endothelial cells and epithelial podocytes. As glomeruli mature, additional basement membrane derived from podocytes is spliced into the fused GBM and loose mesangial matrices condense. The mechanisms for GBM fusion, splicing, and mesangial matrix condensation are not known but might involve intermolecular bond formation between matrix molecules. To test for laminin binding sites, we intravenously injected mouse laminin containing {alpha}1-, ß1-, and {gamma}1-chains into 2-day-old rats. Kidneys were immunolabeled for fluorescence and electron microscopy with domain-specific rat anti-mouse laminin monoclonal antibodies (MAbs), which recognized only mouse and not endogenous rat laminin. Intense labeling for injected laminin was found in mesangial matrices and weaker labeling was seen in GBMs of maturing glomeruli. These patterns persisted for at least 2 weeks after injection. In control newborns receiving sheep IgG, no binding of injected protein was observed and laminin did not bind adult rat glomeruli. To assess which molecular domains might mediate binding to immature glomeruli, three proteolytic laminin fragments were affinity-isolated by MAbs and injected into newborns. These failed to bind glomeruli, presumably owing to enzymatic digestion of binding domains. Alternatively, stable incorporation may require multivalent laminin binding. We conclude that laminin binding sites are transiently present in developing glomeruli and may be functionally important for GBM assembly and mesangial matrix condensation. (J Histochem Cytochem 46:291–300, 1998)

Key Words: glomerular basement, membranes, matrix, laminin, monoclonal antibodies, bamacan, collagen Type IV, entactin, nidogen, perlecan


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Glomerular laminin isoform transitions: errors in metanephric culture are corrected by grafting
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