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Plasma membrane Ca-ATPase isoform expression in human cataractous lenses compared to age-matched clear lenses
Moazez J Marian1, Partha Mukhopadhyay, Douglas Borchman
1Department of Biochemistry and Molecular Biology, University of Louisville, Louisville, Ky, USA.
Abstract:
The plasma membrane calcium ATPase (PMCA) pump is the major mechanism by which calcium is removed from the lens. The aim of this study was to determine if mRNA and proteins levels of PMCA isoforms changed with age or lens opacity. mRNA was quantified using a quantitative real-time reverse transcription polymerase chain reaction assay (RT-PCR). PMCA protein levels were quantified using Western blot analysis. No PMCA mRNA or proteins were detected in human lens fiber cells. The mRNA and protein levels of PMCA1, 3 and 4 in the epithelium of cataractous lenses were similar to those of epithelium from age-matched clear lenses and were also the same in younger lenses. PMCA2 mRNA and protein levels were 1.6-2.5 times higher, respectively, in cataractous lenses compared to age-matched clear lenses. Elevated PMCA2 expression in cataractous lenses might be a compensatory mechanism to overcome higher intracellular calcium levels in cataract.
Insights
Plasma membrane calcium ATPase (PMCA) pump levels in the lens were investigated. Cataractous lenses showed significantly higher PMCA2 expression, suggesting a compensatory response to elevated intracellular calcium.
Area of Science:
- Ophthalmology
- Cell Biology
- Biochemistry
Background:
- The plasma membrane calcium ATPase (PMCA) pump is crucial for calcium extrusion from cells.
- Calcium homeostasis is vital for lens transparency and function.
- Age-related changes and opacity in the lens may affect calcium regulation.
Purpose of the Study:
- To investigate age- and opacity-related changes in PMCA isoform mRNA and protein levels in human lenses.
- To determine the role of PMCA pumps in lens calcium regulation, particularly in cataracts.
Main Methods:
- Quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) for mRNA analysis.
- Western blot analysis for PMCA protein quantification.
- Comparison of PMCA expression in cataractous versus age-matched clear human lenses.
Main Results:
- No PMCA mRNA or protein was detected in human lens fiber cells.
- PMCA1, PMCA3, and PMCA4 mRNA and protein levels were similar in the epithelium of cataractous and clear lenses.
- PMCA2 mRNA and protein levels were significantly elevated (1.6-2.5 times higher) in the epithelium of cataractous lenses compared to age-matched clear lenses.
Conclusions:
- PMCA2 upregulation in cataractous lenses may represent a compensatory mechanism to manage increased intracellular calcium.
- PMCA isoforms exhibit differential expression patterns in the lens epithelium related to cataract formation.
- Understanding PMCA's role in lens calcium homeostasis could offer insights into cataract pathogenesis.
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