Regulation of sarco/endoplasmic Ca2+ -ATPase expression by calcium in human lens cells.
Maintenance of cellular calcium levels is critical to cell function. Loss of calcium homeostasis might be a contributing factor to the development of cataract in the lens. In lens epithelium, calcium is involved in cell signaling and its precise regulation is vital. In this study, we investigated the regulation of sarco/endoplasmic reticulum Ca(2+)-ATPase2b (SERCA2b) and SERCA3 isoform expression in cultured epithelial B-3 cells from human lenses. Both mRNA and membrane proteins samples were collected for semi quantitative RT-PCR using GAPDH as a control. Western blot analyses were performed on membrane samples.Thapsigargin, a SERCA isoform inhibitor which causes increased cytosolic levels elicited dose-and time-dependent up-regulation of SERCA3 at both mRNA and protein levels; SERCA2b expression was unaffected. Both EGTA and actinomycin partially inhibited the thapsigargin-induced SERCA3 up-regulation. These results indicate that the up-regulation of SERCA3 by thapsigargin is dependent on a calcium-mediated pathway that is likely to occur at the transcription level.
Liu
L
L
Department of Ophthalmology and Visual Sciences, University of Louisville, Louisville, KY 40202, USA.
Paterson
C A
CA
Borchman
D
D
D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D002478:Cells, Cultured; D003609:Dactinomycin / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum / Q000201:enzymology*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007903:Lens Capsule, Crystalline / Q000187:drug effects / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019284:Thapsigargin / Q000494:pharmacology
D012333:RNA, Messenger; D003609:Dactinomycin; D019284:Thapsigargin; D000252:Calcium-Transporting ATPases; D002118:Calcium