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Altered Na,K-ATPase pattern in gamma-crystallin mutant mice
Amy Moseley1, Jochen Graw, Nicholas A Delamere
1Department of Ophthalmology and Visual Sciences, School of Medicine, University of Louisville, 301 E. Muhammad Ali Boulevard, Louisville, KY 40292, USA.
Investigative Ophthalmology & Visual Science
|May 1, 2002
Summary
Na,K-adenosine triphosphatase (ATPase) expression is increased in the lenses of cataract mouse mutants. Specifically, alpha2 and alpha3 isoforms are upregulated in the cataractous lens, but not altered in the brain.
Area of Science:
- Ophthalmology
- Molecular Biology
- Biochemistry
Background:
- Cataracts are a leading cause of vision impairment globally.
- Na,K-adenosine triphosphatase (ATPase) plays a crucial role in maintaining cellular ion balance.
- Previous studies indicated elevated Na,K-ATPase activity in specific murine cataract models.
Purpose of the Study:
- To investigate the expression levels of Na,K-adenosine triphosphatase (ATPase) alpha1, alpha2, and alpha3 catalytic subunit polypeptides in the lenses of Cryge(t) and Cryge(ns) mutant mice.
- To determine if changes in Na,K-adenosine triphosphatase (ATPase) expression are specific to the lens or also present in the brain.
Main Methods:
- Membrane material was isolated from the lenses and brains of wild-type, heterozygous, and homozygous mutant mice.
- Microsomal membranes were prepared via centrifugation.
- Na,K-adenosine triphosphatase (ATPase) polypeptides were detected using immunoblot analysis with isoform-specific antibodies.
Main Results:
- Homozygous cataract mutants exhibited dense immunoblot bands for Na,K-adenosine triphosphatase (ATPase) alpha2 and alpha3 isoforms in lens material, not seen in wild-type mice.
- An increased band density for the Na,K-adenosine triphosphatase (ATPase) alpha1 isoform was also observed in homozygous mutant lenses.
- Na,K-adenosine triphosphatase (ATPase) alpha3 was detected in heterozygous mutant lenses, and alpha2 was detected in heterozygous Cryge(t) and to a lesser extent in Cryge(ns) mutant lenses.
- Brain membrane material showed similar Na,K-adenosine triphosphatase (ATPase) subunit expression across all groups.
Conclusions:
- The abundance of Na,K-adenosine triphosphatase (ATPase) polypeptides is elevated in the cataractous mouse lens.
- The expression of Na,K-adenosine triphosphatase (ATPase) alpha2 and alpha3 isoforms is significantly upregulated in the cataractous lens.
- Na,K-adenosine triphosphatase (ATPase) expression in the brain remains unaltered in these cataract models.