May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Expression of Plasma Membrane Calcium ATPase in the Cultured Human Lens Epithelial B-3 Cells
Author Affiliations & Notes
  • H. Li
    Ophthalmology and Visual Sciences, University Louisville, Louisville, KY, United States
  • M.J. Marian
    Ophthalmology and Visual Sciences, University Louisville, Louisville, KY, United States
  • C.A. Paterson
    Ophthalmology and Visual Sciences, University Louisville, Louisville, KY, United States
  • D. Borchman
    Ophthalmology and Visual Sciences, University Louisville, Louisville, KY, United States
  • Footnotes
    Commercial Relationships  H. Li, None; M.J. Marian, None; C.A. Paterson, None; D. Borchman, None.
  • Footnotes
    Support  NIH Grant EY06916
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 4475. doi:
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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)

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Abstract

Abstract: : 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.

Keywords: calcium • cataract • ion transporters 
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