Vascular adventitial cell expression of collagen I messenger ribonucleic acid in anti-glomerular basement membrane antibody-induced crescentic nephritis in the rabbit. A cellular source for interstitial collagen synthesis in inflammatory renal disease.
BACKGROUND
Scarring in the interstitial compartment of the renal cortex heralds a poor prognosis in many forms of renal injury, however, the mechanism through which glomerular inflammation leads to interstitial scarring is not understood. In a model of anti-GBM disease in the rabbit, development of crescentic glomerulonephritis is associated with marked interstitial fibrosis and decreased renal function. We previously demonstrated that collagen accumulation in the model was preceded by increases in collagen I and IV mRNA and that these changes were primarily extraglomerular at early time points when inflammation was predominantly intraglomerular. In order to identify the cellular origins of extraglomerular collagen synthesis in this model, in situ hybridization using an alpha 2(I) procollagen probe was performed.


METHODS
A 602 bp rabbit alpha 2(I) procollagen cDNA was cloned using a PCR strategy and sequenced. The nucleotide sequence of the coding region was 94% identical with the human alpha 2(I) procollagen sequence. Northern blots were performed to define conditions of specific hybridization of the anti-sense riboprobe. Tissue sections from normal rabbit kidneys and from kidneys 4, 5, 7, 10 and 14 days after injection of anti-GBM antibody were hybridized with 35S-labeled sense and anti-sense riboprobes. Cells containing alpha 2(I) mRNA were identified by autoradiography and mRNA abundance was quantitated by grain density.


RESULTS
No specific hybridization was detected with the sense probe at any time. alpha 2(I) mRNA was undetectable with the anti-sense probe in normal kidney sections. In contrast, the anti-sense probe hybridized specifically at all time points after induction of anti-GBM disease. In agreement with previous filter hybridization studies, on day 4, when inflammation was predominantly intraglomerular, cells in the periarterial adventitial compartment of renal cortex hybridized strongly. At later time points, labeling was also present in the interstitial spaces, the periglomerular region, in Bowman's space and in the glomerular tuft itself.


CONCLUSIONS
We conclude that perivascular adventitial cells are among the first to respond to glomerular inflammation and represent a pool of cells that subsequently contribute to interstitial and glomerular scarring.
Department of Internal Medicine, University of Michigan, Ann Arbor.
R
R
Wiggins
M
M
Goyal
S
S
Merritt
P D
PD
Killen
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000494:pharmacology*; D001483:Base Sequence; D001485:Basement Membrane / Q000276:immunology / Q000648:ultrastructure; D015152:Blotting, Northern; D003001:Cloning, Molecular; D003094:Collagen / Q000096:biosynthesis* / Q000235:genetics*; D003238:Connective Tissue / Q000098:blood supply / Q000737:chemistry*; D003239:Connective Tissue Cells; D004247:DNA / Q000032:analysis / Q000235:genetics; D017403:In Situ Hybridization; D007678:Kidney Glomerulus / Q000737:chemistry* / Q000166:cytology* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D009393:Nephritis / Q000209:etiology* / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D011817:Rabbits
D000906:Antibodies; D012333:RNA, Messenger; D003094:Collagen; D004247:DNA