Extracellular matrix formation by epithelial cells from human polycystic kidney cysts in culture |
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Authors: | Giovanni Candiano Rosanna Gusmano Paola Altieri Roberta Bertelli Fabrizio Ginevri Domenico A Coviello Adalberto Sessa Gianluca Caridi Gian Marco Ghiggeri |
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Institution: | 1. Department of Nephrology, G. Gaslini Institute, Genoa, Italy 2. Institute of Biology and Genetics, University of Genoa, Italy 3. Section of Nephrology, Vimercate, Milan, Italy
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Abstract: | Cells from the cysts of patients with autosomal dominant polycystic kidney disease (PKD) were grown in vitro under standard
conditions without the aid of collagen-pretreated surfaces, and both the synthesis and composition of the extracellular matrix
were investigated. At confluence, PKD cells presented the typical features of epithelial cells, but showed a different collagen
composition from fibroblasts. Compared with normal tubular epithelia (NTE), PKD monolayers produced an excess of extracellular
matrix, which accounted for 30% of the total incorporation of 3H] proline, although this value was considerably lower (by a factor of 10) in the case of NTE. Immunohistochemical and electrophoretic
techniques revealed a complex collagen composition in the extracellular matrix which included α(III)]3 and collagen IV. However, part of the collagen components remained unidentified in spite of the fact that they exhibited
a typical Mr of α1(I) and α2(I) in the presence of urea. Immunoprecipitation with monospecific antibodies and Northern blotting with specific probes failed
to recognize α1(I) and α2(I), but demonstrated their presence in fibroblasts. Purification and cyanogen bromide digestion demonstrated a strong interhomology
in fingerprint peptide composition among the uncharacterized collagens synthesized by PKD cells, thus suggesting a common
identity. These observations document a markedly augmented production of extracellular matrix by PKD cultured cells in vitro,
and show the presence of collagens which do not share homologies with the major collagen molecules. A better characterization
of extracellular matrix composition is central to any comprehension of the cystogenetic mechanisms in vivo. |
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Keywords: | Cell cultures Polycystic kidney disease Extracellular matrix Collagens |
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