Measurement of oxygen production by in vitro human and animal lenses with an oxygen electrode.
OBJECTIVE
This paper describes an advantageous method of measuring the activity of the enzyme catalase, which has an important antioxidative role in the lens. This method allows the measurement of catalase in whole lenses.


METHODS
Exposure to UVA (99% UV-A) radiation was used to stress animal and human (Eye Bank) lenses in vitro. The ability of lens catalase to convert H2O2 into O2 was measured directly, using an oxygen electrode and meter. This method is very specific, as catalase is the only enzyme that converts H2O2 to O2.


RESULTS
Catalase in the lenses of humans, rabbits, and squirrels catalyzed the production of O2 from H2O2 very efficiently. The anterior equatorial regions of these lenses were the most active O2 producing areas. More than 95% of lens catalase activity was found in the capsule-epithelium layer. Exposure to UVA radiation, up to approximately 100 J/cm2 in 18 h, strongly inhibited O2 production from 0.77 mM H2O2 by the lenses. Catalase activity decreased with increasing age. Mixed cataractous human lenses produced O2 from H2O2 at only 60% of the rate of normal lenses of similar ages. Nuclear cataracts produced O2 at only 75% of the rate of normal lenses. Alpha-tocopherol (10(-5) M) protected lens catalase activity strongly. Alpha-tocopherol is known to accumulate in and protect against cell membrane peroxidation, and against singlet oxygen formation. These oxidative mechanisms appear to contribute to catalase photoinactivation.


CONCLUSIONS
The method described indicated that catalase is a crucial antioxidative enzyme in the normal lens. Its inactivation could upset the oxidation-reduction balance in the lens and stimulate lens opacification.
Zigman
S
S
University of Rochester School of Medicine and Dentistry, Department of Ophthalmology, NY, USA. zigmanb@ophth.rochester.edu
Schultz
J B
JB
Schultz
M
M
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000502:physiology; D000818:Animals; D002374:Catalase / Q000378:metabolism; D002386:Cataract / Q000188:drug therapy / Q000209:etiology / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D017736:Ion-Selective Electrodes; D007903:Lens Capsule, Crystalline / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D007908:Lens, Crystalline / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D008875:Middle Aged; D010100:Oxygen / Q000378:metabolism*; D011817:Rabbits; D011833:Radiation Injuries, Experimental / Q000188:drug therapy / Q000209:etiology / Q000378:metabolism; D012589:Sciuridae; D014466:Ultraviolet Rays; D014810:Vitamin E / Q000494:pharmacology
D014810:Vitamin E; D006861:Hydrogen Peroxide; D002374:Catalase; D010100:Oxygen