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H. Li, M.J. Marian, C.A. Paterson, D. Borchman; Expression of Plasma Membrane Calcium ATPase in the Cultured Human Lens Epithelial B-3 Cells . Invest. Ophthalmol. Vis. Sci. 2003;44(13):4475.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Plasma membrane calcium ATPase (PMCA) isoforms, PMCA1-4, play important roles in Ca2+ regulation. PMCA1 and PMCA4 are widely expressed while the expression of PMCA2 and PMCA3 exhibit tissue specificity. We tested whether these PMCA isoforms are present in the cultured human lens epithelial B-3 cells (HLE B-3) providing a basis for further studies focused on calcium homeostasis in the lens. Methods: HLE B-3 cells were used to extract mRNA and prepare membrane proteins. Reverse transcription polymerase chain reaction (RT-PCR) was employed to detect mRNA of PMCA isoforms using specific primers. Western blot was used to detect protein of PMCA isoforms using specific anti-PMCA1-4 antibodies. Results: A 429bp fragment with PMCA1 primers, a 557bp fragment with PMCA2 primers and a 849bp fragment with PMCA4 primers were amplified in HLE B-3 cells. All these products have been identified as the expected PMCA1, 2 and 4 products by sequencing analysis. Western blot analysis was used to confirm that PMCA1, 2 and 4 were present with molecular weights of 129, 135 and 129-134KDa, respectively. PMCA3 was not detected at both mRNA and protein levels. Conclusions: PMCA1, PMCA2 and PMCA4 are expressed in cultured human lens epithelial B-3 (HLE B-3) cells. They may play an important role in the regulation of calcium homeostasis in lens epithelial cells. Support: NIH grant EY06916, an unrestricted grant from Research to Prevent Blindness Inc., New York and The Kentucky Lions Eye Foundation.
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