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Updated: Aug 15, 2026
Mucin Agarose Gel Electrophoresis: Western Blotting for High-molecular-weight Glycoproteins
Published on: June 14, 2016
Vitamin A deficiency alters the expression of mucin genes by the rat ocular surface epithelium
M Tei1, S J Spurr-Michaud, A S Tisdale
1Schepens Eye Research Institute, Boston, Massachusetts 02114, USA.
Purpose:
To study effects of depletion of retinoic acid on expression of the mucins ASGP (rMuc4), rMuc5AC, and rMuc1, by the corneal and conjunctival epithelia of the rat.
Methods:
Nineteen-day-old Sprague-Dawley male rats were fed a casein-based vitamin A- deficient diet or casein-based diet with vitamin A as control. Rats from both groups were killed at 1, 3, 5, 13, 15, 18, and 20 weeks after initiation of feeding. Expression of the three mucin genes by the ocular surface epithelium was assayed by reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization.
Results:
In vitamin A-deficient rats, ASGP mRNA was not detected by RT-PCR after 15 weeks of feeding. rMuc5AC mRNA was detected by RT-PCR at 15 weeks, but by 18 and 20 weeks was no longer detectable. By in situ hybridization, ASGP mRNA was localized in the entire ocular surface epithelium after 1 week of feeding, was diminished but detectable above background by 13 weeks, and was not detectable at 20 weeks. rMuc5AC mRNA was detected in the goblet cells of vitamin A- deficient rats by in situ hybridization at 13 weeks, but was lost by 20 weeks, as were identifiable goblet cells. rMuc1 mRNA were detected by RT-PCR through all time points of 1 to 20 weeks in both vitamin A-deficient and control rats, indicating no significant change in rMuc1 mRNA expression with vitamin A deficiency.
Conclusions:
Both the membrane-spanning mucin ASGP (rMuc4) and the secretory mucin rMuc5AC are directly or indirectly regulated by vitamin A in the ocular surface epithelium, whereas the membrane-spanning mucin rMuc1 is not.
