Strong Pasteur effect in rabbit corneal endothelium preserves fluid transport under anaerobic conditions.
1. The hydration and transparency of the mammalian cornea are maintained by a metabolically dependent fluid transport system located in the endothelial cell layer. The purpose of the study was to determine whether this pump activity is dependent upon aerobic or anaerobic metabolism. 2. The ability of the cornea, superfused in vitro with a bicarbonate-Ringer solution containing glucose and adenosine, to maintain normal hydration (thickness) when respiration is inhibited has been examined in intact and de-epithelialized preparations and correlated with glycolytic activity and cellular concentrations of ATP. 3. In respiring intact and de-epithelialized corneas thickness was maintained for periods up to 5 h during superfusion with the control Ringer solution. 4. KCN (10(-3) M) or antimycin A (10(-6) M) caused the intact cornea to swell at 15 +/- 3 microns h-1, whereas the de-epithelialized tissue maintained normal thickness under these conditions. This swelling of the intact tissue appears to be due to the osmotic effect of increased epithelial lactate production under anaerobic conditions. 5. Pre-swollen de-epithelialized corneas deturgesced fully to original thickness at a rate of 43 +/- 7 microns h-1 under aerobic conditions, but with KCN or antimycin they deturgesced at only 65% of that rate and generally failed to return to their original thickness. 6. Ouabain (10(-4) M) caused de-epithelialized corneas to swell in the presence of KCN or antimycin, as it did under aerobic conditions, showing that maintenance of hydration or deturgescence are pump-dependent processes under both conditions. 7. Lactate production was markedly stimulated by KCN or antimycin in intact and de-epithelialized preparations, but not in the stroma alone. The magnitude of the Pasteur effect was calculated to be 5-fold in the endothelium and 2.5-fold in the epithelium. Ouabain inhibited anaerobic lactate production in the endothelium by 50%. 8. ATP content of the epithelium following 5 h superfusion was 22.0 nmol cm-2 in control (aerobic) corneas, but fell to 1.9 nmol cm-2 in the presence of 10(-3) M-KCN, whereas the endothelial value fell only from 1.1 to 0.7 nmol cm-2 under these conditions. 9. Omission of glucose from the medium containing KCN or antimycin caused immediate swelling of tissues and a rapid decline of ATP content to less than 1% of that in control conditions.(ABSTRACT TRUNCATED AT 400 WORDS)
Riley
M V
MV
Eye Research Institute, Oakland University, Rochester, MI 48309.
Winkler
B S
BS
D000255:Adenosine Triphosphate / Q000378:metabolism; D000693:Anaerobiosis / Q000502:physiology; D000818:Animals; D000968:Antimycin A / Q000494:pharmacology; D001693:Biological Transport, Active / Q000187:drug effects; D004728:Endothelium, Corneal / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D006019:Glycolysis; D007773:Lactates / Q000378:metabolism; D010042:Ouabain / Q000494:pharmacology; D011190:Potassium Cyanide / Q000494:pharmacology; D011817:Rabbits; D013997:Time Factors
D007773:Lactates; D010042:Ouabain; D000968:Antimycin A; D000255:Adenosine Triphosphate; D005947:Glucose; D011190:Potassium Cyanide