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Na,K-ATPase response to osmotic stress in primary dog lens epithelial cells.
Invest Ophthalmol Vis Sci. 1995 Jan; 36(1):88-94.IO

Abstract

PURPOSE

Na,K-ATPase activity increases in lens cells exposed to hypertonic stress. To test whether the increase in activity involves stimulation of Na,K-ATPase expression, dog lens epithelial cells were subjected to hypertonic stress, and the time course of Na,K-ATPase protein and mRNA response was measured.

METHODS

Primary cultures of dog lens epithelial cells were maintained in isotonic or hypertonic media over the course of several days. Rubidium-86 uptake measurements, immunoreactive protein, and northern blot analysis were performed.

RESULTS

Dog lens epithelial cells exposed to hypertonic stress from culture medium supplemented with 150 mM NaCl or 250 mM cellobiose showed a twofold increase in Na,K-ATPase activity. The increase in activity was blocked by cycloheximide and was reversible when the cells were returned to isotonic medium. This activity was unaffected by the aldose reductase inhibitor, tolrestat. Na,K-ATPase protein and mRNA levels increased in cells exposed to medium containing 150 mM NaCl. Northern blot analysis showed that the alpha-1 and beta-1 mRNA levels increased as early as 6 hours and maximally increased 1.5-fold to twofold by 12 to 24 hours.

CONCLUSIONS

Elevation of Na,K-ATPase activity in dog lens epithelial cells exposed to hypertonic stress was associated with increased expression of Na,K-ATPase subunit mRNAs and was dependent on protein synthesis. These results suggest that upregulation of the enzyme activity is the result of an induction of Na,K-ATPase.

Authors+Show Affiliations

National Eye Institute, National Institutes of Health, Bethesda, Maryland 20892.No affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

7822162

Citation

Old, S E., et al. "Na,K-ATPase Response to Osmotic Stress in Primary Dog Lens Epithelial Cells." Investigative Ophthalmology & Visual Science, vol. 36, no. 1, 1995, pp. 88-94.
Old SE, Carper DA, Hohman TC. Na,K-ATPase response to osmotic stress in primary dog lens epithelial cells. Invest Ophthalmol Vis Sci. 1995;36(1):88-94.
Old, S. E., Carper, D. A., & Hohman, T. C. (1995). Na,K-ATPase response to osmotic stress in primary dog lens epithelial cells. Investigative Ophthalmology & Visual Science, 36(1), 88-94.
Old SE, Carper DA, Hohman TC. Na,K-ATPase Response to Osmotic Stress in Primary Dog Lens Epithelial Cells. Invest Ophthalmol Vis Sci. 1995;36(1):88-94. PubMed PMID: 7822162.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Na,K-ATPase response to osmotic stress in primary dog lens epithelial cells. AU - Old,S E, AU - Carper,D A, AU - Hohman,T C, PY - 1995/1/1/pubmed PY - 1995/1/1/medline PY - 1995/1/1/entrez SP - 88 EP - 94 JF - Investigative ophthalmology & visual science JO - Invest Ophthalmol Vis Sci VL - 36 IS - 1 N2 - PURPOSE: Na,K-ATPase activity increases in lens cells exposed to hypertonic stress. To test whether the increase in activity involves stimulation of Na,K-ATPase expression, dog lens epithelial cells were subjected to hypertonic stress, and the time course of Na,K-ATPase protein and mRNA response was measured. METHODS: Primary cultures of dog lens epithelial cells were maintained in isotonic or hypertonic media over the course of several days. Rubidium-86 uptake measurements, immunoreactive protein, and northern blot analysis were performed. RESULTS: Dog lens epithelial cells exposed to hypertonic stress from culture medium supplemented with 150 mM NaCl or 250 mM cellobiose showed a twofold increase in Na,K-ATPase activity. The increase in activity was blocked by cycloheximide and was reversible when the cells were returned to isotonic medium. This activity was unaffected by the aldose reductase inhibitor, tolrestat. Na,K-ATPase protein and mRNA levels increased in cells exposed to medium containing 150 mM NaCl. Northern blot analysis showed that the alpha-1 and beta-1 mRNA levels increased as early as 6 hours and maximally increased 1.5-fold to twofold by 12 to 24 hours. CONCLUSIONS: Elevation of Na,K-ATPase activity in dog lens epithelial cells exposed to hypertonic stress was associated with increased expression of Na,K-ATPase subunit mRNAs and was dependent on protein synthesis. These results suggest that upregulation of the enzyme activity is the result of an induction of Na,K-ATPase. SN - 0146-0404 UR - https://www.unboundmedicine.com/medline/citation/7822162/NaK_ATPase_response_to_osmotic_stress_in_primary_dog_lens_epithelial_cells_ L2 - https://iovs.arvojournals.org/article.aspx?volume=36&issue=1&page=88 DB - PRIME DP - Unbound Medicine ER -