Transforming growth factor β blocks cystogenesis by MDCK epithelium in vitro by enhancing the paracellular flux: Implication of collagen V

Paola Altieri, Olga Zegarra Moran, Luis Juan Vicente Galietta, Laura Torri Tarelli, Adalberto Sessa, Gian Marco Ghiggeri

Research output: Contribution to journalArticle

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Abstract

Transforming growth factor β (TGFβ) determines a nearly complete inhibition of cystogenesis by MDCK cells grown in collagen I-enriched matrices in vitro. In order to elucidate the mechanism implicated in this phenomenon, we performed a series of experiments aimed at discovering a relevant role of extracellular matrix. TGFβ (2 ng/ml) played a marked stimulatory effect on the expression of extracellular matrix by MDCK with a selective effect on collagen V (three to fourfold increase of protein and mRNA) and in parallel inhibited cystogenesis by 95%. Cotreatment with TGFβ and anti-collagen V antibodies restored a normal cystogenesis. In analogy, when MDCK cells were grown in three-dimensional matrices containing collagen I and minor (10%) amounts of collagen V, cystogenesis was once again inhibited by 95%. To characterize the molecular mechanism activated by TGFβ and collagen V, we looked at the electrophysiological characteristics of MDCK monolayers and found a drastic fall of transepithelial electrical resistance (TER) in both conditions. In parallel with the decrease in TER, TGFβ and collagen V also induced the leakage of two high molecular weight tracers, i.e., [3H]-inulin and 150 kD FITC-Dextran, suggesting a perturbation of the paracellular permeability. Finally, TGFβ at the relevant concentration did not stimulate apoptosis in our cellular model, as judged by propidium iodide staining and by in situ end labeling of DNA fragments. These observations suggest that TGFβ inhibits cystogenesis by MDCK cells in vitro by altering the collagenic composition of the three-dimensional milieu where MDCK cells grow and form cysts. The molecular mechanism responsible for inhibition of cystogenesis is the increase of paracellular flux which overcomes the active transport of solutes and water inside cysts.

Original languageEnglish
Pages (from-to)214-223
Number of pages10
JournalJournal of Cellular Physiology
Volume177
Issue number2
DOIs
Publication statusPublished - Nov 1998

Fingerprint

Transforming Growth Factors
Collagen
Epithelium
Madin Darby Canine Kidney Cells
Fluxes
Acoustic impedance
Electric Impedance
Extracellular Matrix
Cysts
Inulin
Propidium
Active Biological Transport
In Vitro Techniques
Labeling
Monolayers
Permeability
Molecular Weight
Molecular weight
Cells
Apoptosis

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Transforming growth factor β blocks cystogenesis by MDCK epithelium in vitro by enhancing the paracellular flux : Implication of collagen V. / Altieri, Paola; Moran, Olga Zegarra; Galietta, Luis Juan Vicente; Tarelli, Laura Torri; Sessa, Adalberto; Ghiggeri, Gian Marco.

In: Journal of Cellular Physiology, Vol. 177, No. 2, 11.1998, p. 214-223.

Research output: Contribution to journalArticle

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