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  • aldose reductase mRNA  (3)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Molecular and cellular biochemistry 115 (1992), S. 27-34 
    ISSN: 1573-4919
    Keywords: aldose reductase mRNA ; galactose cataracts ; lens epithelium ; laser densitometry ; Northern blots ; in situ hybridization ; gas chromatography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Aldose reductase (AR), a major enzyme in the polyol pathway, is thought to be responsible for accumulation of polyols in lenses exposed to high doses of galactose or glucose, and it may be linked to some of the complications found in diabetes. In this report we examined the level of expression of AR mRNA in lens epithelia undergoing development of galactose cataracts in vivo. The AR mRNA was quantitated by Northern blot hybridization with a [35S]-RNA transcript from a previously described AR cDNA clone. This was done on normal lens epithelia and on epithelia from lens of rats fed a diet of Purina Chow containing 50% galactose for periods of from 6 hr to 20 days. We found AR mRNA to elevate to about 5-fold the control levels by 12–24 hr on galactose, then decrease to the control levels by day 4. The increase in AR mRNA appears to be transitory. The high abundance in AR mRNA by 24 hr on galactose was confirmed by in situ hybridization. At later periods, from 8 to 20 days on galactose, a slow increase in AR mRNA took effect, as we have previously reported. Changes in the levels of galactose and dulcitol between 0 and 96 hr were also quantitated by gas chromatography, showing that there was a significant increase in both galactose and dulcitol occurring throughout the experimental period. The induction in mRNA accumulation in the lens epithelia within the first 12–24 hr does not appear to be specific for AR mRNA, but it is widespread and affects at least three other lens genes coding for: the γ-crystallin, the lens intrinsic membrane protein MP26 and c-myc.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Molecular and cellular biochemistry 131 (1994), S. 35-41 
    ISSN: 1573-4919
    Keywords: galactose cataracts ; aldose reductase mRNA ; in situ hybridization ; fiber cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Aldose reductase (AR), an enzyme of the polyol pathway, has been implicated in the pathogenesis of diabetic and galactosemic cataracts. AR mRNA is a specific transcript of the lens epithelial cells. However, in addition to its presence at high levels in the epithelial cells at the equator, it is also found at significant concentrations in fiber cells at the lens bow. In this study we extended our previous work, and examined the distribution of AR mRNA by in situ hybridization in lens of rats maintained on a 50% galactose diet for up to 20 days, then reversed to a normal diet for an additional 20 days. It was found that within 8 days on galactose the posterior AR mRNA dissipated, with no clear increase in this transcript in the lens epithelial cells. By 16 to 17 days on galactose, the surviving fiber cells in the equatorial region appeared to express AR mRNA; these cells in the normal lens contain insignificant amounts of AR mRNA, as we have previously demonstrated. AR mRNA in the fiber cells of the same base length as that found in the control, indicating that the AR mRNA in these cells may not represent a degraded transcript. Upon reversal of the cataracts, the posterior AR mRNA re-accumulates, and the fiber cells at the equator appear to have lost the AR mRNA that accumulated during the period of exposure to galactose. The data demonstrate that during formation of galactose-cataract, surviving cortical fiber cells express significant levels of AR mRNA, while following withdrawal from galactose these fiber cells lose that activity.
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  • 3
    ISSN: 1573-4919
    Keywords: aldose reductase mRNA ; cytoplasmic dot hybridization ; lens mRNA ; reversed cataracts ; galactose cataracts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Aldose reductase (AR) mRNA concentration in rat lens was quantitated by hybridization of a RNA transcript from a previously described AR cDNA clone to mRNA found in epithelial and cortical cytosols. This was done on normal rat lens and on lens initially made cataractous by feeding of a diet of Purina Chow containing 50% galactose, followed by reversal of the cataracts due to the removal of the galactose from the diet. Recent data from this laboratory has shown that AR mRNA was increased in lens epithelial cells upon administration of galactose; while in the cortex it was reduced to insignificant levels when fiber cell damage became extensive by day 20 on galactose. Present data reveals that, upon removal of galactose from the diet, the lens epithelial AR mRNA was gradually reduced from the high levels found at day 20 of galactose feeding to low levels by day 30 of reversal. On the other hand, the cortex exhibited an initial sudden increase in AR mRNA at days 1 to 6 of reversal and by day 30 it was reduced to levels below those found in the untreated lens. DNA content in the epithelium also began to decrease to normal levels by day 16 following reversal of the cataracts. The data demonstrate that the concentration of AR mRNA in lens of reversed cataracts appears to faithfully reflect the loss of epithelial cellular need for AR mRNA in favor of enhanced differentiation of epithelial cells to secondary fiber cells.
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