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Related Experiment Videos

Ascorbate regeneration in bovine ocular tissues by NADH-dependent semidehydroascorbate reductase.

M Khatami, L E Roel, W Li

    Experimental Eye Research
    |August 1, 1986
    PubMed
    Summary

    Bovine ocular tissues possess a semidehydroascorbate reductase (SDR) system that regenerates ascorbate. This NADH-dependent enzyme activity is highest in the retina and sensitive to temperature and chemical treatments.

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    Area of Science:

    • Biochemistry
    • Ophthalmology
    • Enzymology

    Background:

    • Ascorbate (vitamin C) readily autoxidizes in vitro but is maintained in its reduced form in vivo.
    • This suggests an endogenous mechanism for ascorbate regeneration exists within biological systems.

    Purpose of the Study:

    • To investigate the presence and activity of a semidehydroascorbate reductase (SDR) system in bovine ocular tissues.
    • To determine the distribution and characteristics of SDR activity within the eye.

    Main Methods:

    • Extraction of bovine ocular tissues using Triton X-100.
    • Assay of NADH-dependent reductase activity to measure semidehydroascorbate reductase (SDR) activity.
    • Evaluation of SDR activity under varying conditions (temperature, chemical inhibition).

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    Main Results:

    • Semidehydroascorbate reductase (SDR) activity was demonstrated in bovine ocular tissues.
    • Highest SDR activity was found in retinal extracts, followed by pigment epithelium-choroid and ciliary body.
    • Lens and aqueous fluid showed minimal to no SDR activity.
    • Enzyme activity was sensitive to freezing/thawing, boiling, low temperatures (5°C), and N-ethylmaleimide treatment.

    Conclusions:

    • Bovine ocular tissues possess an NADH-dependent semidehydroascorbate reductase (SDR) system crucial for ascorbate regeneration.
    • The retina is a primary site of this regeneration activity.
    • SDR activity is influenced by environmental factors and specific chemical modifiers, indicating its enzymatic nature and potential regulatory mechanisms.