Hypertonic stress increases NaK ATPase, taurine, and myoinositol in human lens and retinal pigment epithelial cultures.
OBJECTIVE
Recent evidence suggests that taurine and myoinositol may serve as organic osmolytes in a number of cells, including lens and retinal pigment epithelia, but the mechanism for their increased accumulation in response to hypertonic stress is not known. To assess whether NaK ATPase contributed to the elevated levels of taurine and myoinositol in cells exposed to hypertonic media, we measured the activity of NaK ATPase, which is known to be implicated in the transport of these substances, in human lens and retinal pigment epithelia cultured in isotonic and hypertonic media.


METHODS
Primary cultures of human lens epithelial (HLE) and human retinal pigment epithelial (HRPE) cells were maintained in isotonic and hypertonic media for varying periods of time, and the activity of NaK ATPase and the levels of taurine and myoinositol were measured in cells cultured under two different conditions. The possible involvement of the transport enzyme in the accumulation of the two osmolytes was also investigated by inhibiting the enzyme with ouabain.


RESULTS
When primary cultures of HLE and HRPE were exposed to hypertonic medium containing NaCl (600 mOsm) or cellobiose (500m Osm) for 72 hours, the concentration of taurine and myoinositol in HLE cells increased by 218% and 558% of control, respectively, in NaCl medium, whereas the corresponding increases in cellobiose medium were 147% and 439%. In HRPE cells, the increase in myoinositol levels in the two hypertonic media was more dramatic than that in taurine. Concomitant with the increase in the concentration of the osmolytes, there was an increase in NaK ATPase activity in both cell types. Although the accumulation of taurine in HLE cells in hypertonic media in a 6-hour culture was essentially prevented by 10(-8) mmol/l ouabain, myoinositol levels were affected to a lesser, but still significant, extent. In HRPE cells, which were cultured for 24 hrs in the presence of 10(-6) mmol/l ouabain, there was a more direct correlation between the inhibition of NaK ATPase and the decreased accumulation of taurine and myoinositol in the hypertonic media.


CONCLUSIONS
Although the exact mechanism by which NaK ATPase activity increases in response to hypertonic stress remains to be established, the increased activity of the enzyme is related to the enhanced accumulation of the organic osmolytes, taurine, and myoinositol, in HLE and HRPE cells cultured in hypertonic medium.
Yokoyama
T
T
Eye Research Institute, Oakland University, Rochester, Michigan.
Lin
L R
LR
Chakrapani
B
B
Reddy
V N
VN
D002475:Cellobiose / Q000494:pharmacology*; D002478:Cells, Cultured; D003470:Culture Media; D004848:Epithelium / Q000187:drug effects / Q000201:enzymology; D006801:Humans; D006982:Hypertonic Solutions; D007294:Inositol / Q000378:metabolism*; D007552:Isotonic Solutions; D007908:Lens, Crystalline / Q000187:drug effects* / Q000201:enzymology; D009994:Osmolar Concentration; D010042:Ouabain / Q000494:pharmacology; D010857:Pigment Epithelium of Eye / Q000187:drug effects* / Q000201:enzymology; D012965:Sodium Chloride / Q000494:pharmacology*; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013312:Stress, Physiological; D013654:Taurine / Q000378:metabolism*
D003470:Culture Media; D006982:Hypertonic Solutions; D007552:Isotonic Solutions; D002475:Cellobiose; D013654:Taurine; D012965:Sodium Chloride; D007294:Inositol; D010042:Ouabain; D000254:Sodium-Potassium-Exchanging ATPase