title: Morphology and migration of podocytes are affected by CD151 levels.
abstract: CD151, a member of the tetraspanin family of membrane proteins, is crucially involved in the formation of the glomerular filtration barrier in humans and mice. However, the role of CD151 in podocytes has not been investigated so far. In the present study, we utilized a conditionally immortalized mouse podocyte cell line to characterize CD151 in podocytes and to examine the consequences of manipulating CD151 expression levels. Mouse podocytes endogenously express CD151 as determined by RT-PCR and Western blotting. GFP-CD151 fusion protein localized to the cell membrane, to cell protrusions and cell-cell contacts, colocalizing with actin, β(1)-integrin, zonula occludens-1, and CD9. The expression of GFP-CD151 in cultured podocytes resulted in a marked increase in the presence of thin arborized protrusions (TAPs). TAPs are distinct from filopodia by increased length, protein composition, branched morphology, and slower dynamics. Furthermore, the migration rate of pEGFP-CD151-transfected podocytes was reduced in a wound assay. Fluorescence recovery after photo bleaching measurements revealed a half-time of 3 s for GFP-CD151 consistent with a high mobility of CD151 in the membrane and cytosol. CD151 knockdown in podocytes reduced β(1)-integrin expression and podocyte cell area, indicating diminished adherence and/or spreading. Our results indicate that CD151 importantly modulates podocyte function.
authors.affiliation: Department of Anatomy and Cell Biology, Universitätsmedizin Greifswald, Friedrich-Loeffler-Str. 23c, 17487 Greifswald, Germany.
authors.forename: Antje
authors.initials: A
authors.lastname: Blumenthal
authors.forename: Jürgen
authors.initials: J
authors.lastname: Giebel
authors.forename: Ramesh
authors.initials: R
authors.lastname: Ummanni
authors.forename: Rabea
authors.initials: R
authors.lastname: Schlüter
authors.forename: Karlhans
authors.initials: K
authors.lastname: Endlich
authors.forename: Nicole
authors.initials: N
authors.lastname: Endlich
mesh_terms: D000199:Actins / Q000378:metabolism; D000230:Adenocarcinoma; D000818:Animals; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology*; D036681:Fluorescence Recovery After Photobleaching; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism; D007365:Intercellular Junctions / Q000378:metabolism; D007680:Kidney Neoplasms; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D010750:Phosphoproteins / Q000378:metabolism; D050199:Podocytes / Q000166:cytology* / Q000502:physiology*; D060185:Tetraspanin 24 / Q000235:genetics / Q000378:metabolism*; D060245:Tetraspanin 29 / Q000378:metabolism; D059470:Tetraspanins / Q000378:metabolism; D062826:Zonula Occludens-1 Protein
chemical_list: D000199:Actins; C554715:CD151 protein, human; C554869:CD9 protein, human; C554716:Cd151 protein, mouse; C554870:Cd9 protein, mouse; D019012:Integrin beta1; D008565:Membrane Proteins; D010750:Phosphoproteins; C569042:TJP1 protein, human; D060185:Tetraspanin 24; D060245:Tetraspanin 29; D059470:Tetraspanins; C569043:Tjp1 protein, mouse; D062826:Zonula Occludens-1 Protein; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins