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| POLYMERASE CHAIN-REACTION AND FOCAL CONTACT FORMATION INDICATE INTEGRIN EXPRESSION IN MESANGIAL CELLS |
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| Author(s): PETERMANN A, FEES H, GRENZ H, GOODMAN SL, STERZEL RB |
| Source: KIDNEY INTERNATIONAL Volume: 44 Issue: 5 Pages: 997-1005 Published: NOV 1993 |
| Times Cited: 25 References: 53 |
| Abstract: Cultured kidney glomerular mesangial cells (MCs) allow the role of extracellular matrix (ECM) and growth factors in glomerular inflammatory disease to be studied. To investigate the potential of MCs to interact with matrix components, the expression of integrin mRNA in cultured MCs was examined by polymerase chain reaction (PCR), by Northern blotting and by immunofluorescence. In addition, the effect of matrix substrates on mRNA expression was assessed by PCR. Northern blots with cDNA probes to integrin alpha-chains revealed that MCs expressed alpha1, alpha3 and alpha5 integrin mRNA. alpha1 and alpha3 were the major messages. No alpha2, alpha4 or alpha6 were detectable. RT-PCR revealed that alpha2 and alpha6 were also expressed at low levels. The control cells, HT1080, expressed alpha2, alpha3, alpha4, alpha5 and alpha6 mRNA, and Rugli expressed alpha1, alpha3, and alpha5, supporting previous studies. Immunocytochemistry confirmed that alpha1beta1, alpha2beta1, alpha3beta1 and alpha5beta1 integrins were expressed and that they were concentrated into focal adhesions (alpha1beta1 on type I collagen and laminin; alpha2beta1 on type I collagen; alpha3beta1 on type I collagen, laminin and fibronectin; alpha5beta1 on fibronectin). Alpha6beta1 was not detected in focal contacts. Attachment, spreading, and formation of talin and integrin containing focal contacts still occurred when endogenous protein synthesis was blocked with 30 mug . ml-1 cycloheximide. Variation of substrate did not lead to a rapid degradation of integrin alpha-chain mRNA as assessed by RT-PCR. These results provide a basis for studying the regulation of interactions between MCs and extracellular matrix mediated by integrins. |
| Document Type: Article |
| Language: English |
Addresses:
1. E MERCK AG, PHARMA F+E, PRECLIN PHARMACEUT RES, FRANKFURTERSTR 250, W-6100 DARMSTADT 1, GERMANY
2. UNIV ERLANGEN NURNBERG, MED KLIN 4, NEPHROL RES LAB, W-8520 ERLANGEN, GERMANY |
| Publisher: BLACKWELL SCIENCE INC, 238 MAIN ST, CAMBRIDGE, MA 02142 |
| Subject Category: Urology & Nephrology |
| IDS Number: MB448 |
| ISSN: 0085-2538 |
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