November 1989
Volume 30, Issue 11
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Articles  |   November 1989
A comparative study of ascorbic acid entry into aqueous and vitreous humors of the rat and guinea pig.
Author Affiliations
  • J DiMattio
    Department of Physiology and Biophysics, New York University School of Medicine, New York.
Investigative Ophthalmology & Visual Science November 1989, Vol.30, 2320-2331. doi:
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      J DiMattio; A comparative study of ascorbic acid entry into aqueous and vitreous humors of the rat and guinea pig.. Invest. Ophthalmol. Vis. Sci. 1989;30(11):2320-2331.

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Abstract

The transport rates of radiolabeled ascorbic acid and dehydroascorbic acid, as well as, labeled 3-O-methyl-D-glucose and L-glucose from a central plasma compartment into aqueous and vitreous humors and cerebrospinal fluid were studied in vivo. Normal, male albino Sprague-Dawley rats and English Short Haired guinea pigs were used to explore the mechanism of ascorbic acid entry into ocular humors in a species that can produce ascorbate (the rat) and one that cannot and, like humans, is dependent on dietary sources (the guinea pig). In vivo kinetic studies allowed for the calculation of entry rate constants, Ki (min-1), in double-labeled experiments using L-glucose as an internal passive control. Parallel TLC chromatographic studies were performed to monitor intraocular labeled molecules deriving from the plasma-introduced test molecule. In addition, resting levels of ascorbic acid and D-glucose were determined in order to obtain more reliable data than previously available. Resting levels of D-glucose revealed a consistent pattern of lower levels in aqueous and vitreous humors and CSF than found in plasma for both rat and guinea pig. However, ascorbate levels differed significantly, with the guinea pig demonstrating high ascorbate levels in the aforementioned humors: 58, 77 and 22, respectively, times the circulating plasma value of 0.2 +/- 0.2 mg/dl. In contrast, the rat, like the guinea pig, had low plasma ascorbate levels (3.3 +/- 0.8 mg/dl) compared to glucose (162 +/- 8 mg/dl), with even lower aqueous and vitreous values in a pattern similar to that of D-glucose. In vivo aqueous, vitreous and CSF transport results from the guinea pig indicate active transport mechanisms for ascorbic acid that prefer the ascorbate over the dehydroascorbate moiety and are probably different from the carrier-facilitated diffusion mechanisms for D-glucose, which do not move molecules against a concentration gradient. TLC studies, performed under nitrogen, revealed that only (14C)-ascorbic acid was present in aqueous or vitreous humors regardless of whether the radiolabeled pulse was of ascorbic or dehydroascorbic acid. The rat demonstrated little or no carrier involvement, with ascorbic acid crossing into ocular humors at rates very close to those of L-glucose, which is similar in size and is considered to cross the barriers studied via passive diffusion. Saturation studies with unlabeled glucose and glucose inhibitor drugs phloretin (10(-3) M) and phloridzin (10(-1)) had no apparent effect on ocular entry rates. Dehydroascorbic acid movement was also found to be passive.(ABSTRACT TRUNCATED AT 400 WORDS)

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