PMID:15292044

Citation	Ziehr, J, Sheibani, N and Sorenson, CM (2004) Alterations in cell-adhesive and migratory properties of proximal tubule and collecting duct cells from bcl-2 -/- mice. Am. J. Physiol. Renal Physiol. 287:F1154-63
Abstract	Bcl-2 protects cells from apoptosis initiated by a variety of stimuli including loss of cell adhesion. Bcl-2 -/- mice develop renal hypoplastic/cystic dysplasia with renal cyst formation coinciding with renal maturation in normal mice. To gain a better understanding of the role cell-adhesive mechanisms play during renal maturation, we generated proximal tubule and collecting duct cell lines from postnatal day 10 (P10) and P20 bcl-2 +/+ and bcl-2 -/- mice. Very little is known about the role cell-adhesive and migratory mechanisms play during renal maturation. We observed that modulation of cell-adhesive properties, which normally occur in a nephron segment-specific manner during renal maturation, and cell migration were altered in cells from bcl-2 -/- mice. Enhanced migration of bcl-2 -/- proximal tubule cells in a scratch wound assay was completely inhibited by incubation with PP1 (Src inhibitor) and moderately affected by incubation with SB-203580 (p38 inhibitor). These cells expressed increased levels of fibronectin and had numerous central focal adhesions. P20 bcl-2 -/- proximal tubule cells adhered to fibronectin but adhered poorly to collagen, vitronectin, or laminin. Collecting duct cells, similar to proximal tubule cells from bcl-2 -/- mice, demonstrated enhanced migration in a scratch wound assay that was inhibited by incubation with PP1. Migration of these cells was moderately affected by incubation with PD-98059 (MEK inhibitor) or LY-294002 (PI3 kinase inhibitor), whereas incubation with SB-203580 had no effect. P10 bcl-2 -/- collecting duct cells also expressed increased levels of fibronectin but decreased levels of thrombospondin-1 and demonstrated precocious binding to fibronectin and vitronectin compared with bcl-2 +/+ cells. The ability of P20 bcl-2 +/+ collecting duct cells to adhere to fibronectin and vitronectin corresponded with a decline in thrombospondin-1 expression. Therefore, alterations in cell-adhesive and migratory characteristics may be an early indicator of aberrant renal epithelial cell differentiation.
Links	PubMed Online version:10.1152/ajprenal.00129.2004
Keywords	Animals; Cell Adhesion/physiology; Cell Communication; Cell Differentiation; Cell Movement/physiology; Cells, Cultured; Extracellular Matrix/physiology; Extracellular Matrix Proteins/metabolism; Extracellular Signal-Regulated MAP Kinases/metabolism; Fibronectins/metabolism; Fibronectins/secretion; Integrin alpha5/analysis; Kidney Tubules, Collecting/cytology; Kidney Tubules, Collecting/physiology; Kidney Tubules, Proximal/cytology; Kidney Tubules, Proximal/physiology; Mice; Mice, Knockout; Phosphatidylinositol 3-Kinases/metabolism; Proto-Oncogene Proteins c-bcl-2/deficiency; Proto-Oncogene Proteins c-bcl-2/physiology; Signal Transduction; Thrombospondin 1/secretion; p38 Mitogen-Activated Protein Kinases/metabolism; src-Family Kinases/metabolism
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