Cultured peritoneal mesothelial cells exhibit apical primary cilia.
This study examined primary cilia on cultured human and rabbit peritoneal mesothelial cells (PMC) and investigated factors that influence ciliary expression. Primary cilia were examined with indirect immunocytochemistry, laser scanning confocal microscopy and scanning electron microscopy. Ciliary expression was evaluated in cultures with or without l-cysteine (0.25 mM) or exposure to Ca(2+)-free Krebs-Ringer solution supplemented with EGTA, 0.5 mM. This treatment disrupted cell monolayer integrity. Cilia were counted and normalized to total cell counts using NIH image. Primary cilia were identified on both human and rabbit PMC. Cells treated with l-cysteine expressed significantly more cilia compared with monolayers deprived of l-cysteine. Exposure to Ca(2+)-free Krebs-Ringer solution significantly reduced cilia (5.9+/-1.0%, n=7). Although ciliary expression could be augmented with l-cysteine, approximately 60% of human PMC and 84% of rabbit PMC did not exhibit cilia. Together, these data show that monolayers of PMC express apical cilia that can be augmented with l-cysteine, independently of increased cell density.
Department of Medical and Surgical Sciences, Dunedin School of Medicine, University of Otago, PO Box 913, Dunedin, New Zealand. sdbird@hotmail.com
Stephen D
SD
Bird
Michael
M
Legge
Robert J
RJ
Walker
D000818:Animals; D002478:Cells, Cultured; D002923:Cilia / Q000648:ultrastructure*; D003545:Cysteine / Q000737:chemistry; D004533:Egtazic Acid / Q000737:chemistry; D004847:Epithelial Cells / Q000648:ultrastructure*; D006801:Humans; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D008871:Microvilli / Q000648:ultrastructure*; D010529:Peritoneal Cavity / Q000166:cytology*; D011817:Rabbits
D004533:Egtazic Acid; D003545:Cysteine