Asymmetric origins of the mature glomerular basement membrane

• Published in: Journal of cellular physiology Vol. 157; no. 1; p. 169
• Main Authors: Lee, L K, Pollock, A S, Lovett, D H
• Format: Journal Article
• Language: English
• Published: United States 01.10.1993
• Subjects: Animals; Base Sequence; Basement Membrane - metabolism; Basement Membrane - ultrastructure; In Situ Hybridization; Kidney Glomerulus - cytology; Kidney Glomerulus - metabolism; Kidney Glomerulus - ultrastructure; Molecular Sequence Data; Oligonucleotide Probes - genetics; Proteins - genetics; Rats; Rats, Sprague-Dawley; RNA, Messenger - metabolism; Tissue Distribution; Transcription, Genetic
• ISSN: 0021-9541
• DOI: 10.1002/jcp.1041570122

Renal plasma filtration is a critical physiologic function that depends upon the precise composition and arrangement of the constituent extracellular matrix proteins within the glomerular basement membrane (GBM). The GBM develops during renal embryogenesis by the fusion of discrete basement membranes produced independently by endothelial and visceral epithelial cells, and, possibly from matrix secreted by the mesangial cells. In the mature animal, however, the epithelial cell has generally been accepted as the sole source of all GBM constituent proteins. Although the final structures and distributions of the component proteins have been defined by histochemical techniques, the individual contributions of the three resident glomerular cell types to the maintenance and turnover of the mature GBM remain uncertain. We report the application of a new technique, in situ reverse transcription (ISRT), for the localization of RNA transcripts of nine major GBM protein components within the closely apposed cells of the glomerulus. Using this technique, we demonstrate that in normal adult rat glomeruli the RNA transcripts for heparan sulfate proteoglycan and the laminin-S chain are primarily expressed by visceral epithelial cells, while Type IV alpha-1 and alpha-2 collagen transcripts were restricted to the endothelial cells in a heterogeneous pattern. RNA transcripts for entactin and the laminin-A and -B2 chains were expressed by all three glomerular cell types, while laminin-B1 and fibronectin transcripts were limited to the mesangium. These findings demonstrate that GBM synthesis in the mature animal is not restricted to the epithelial cell and that all intrinsic glomerular cells contribute to the production of GBM protein components. The ISRT technique also provided the additional, and unexpected, finding that appreciable synthetic heterogeneity exists within individual glomerular cell types.