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RM Corrales, M Calonge, V Saez, A Mayo, JM Herreras, FJ Chaves; Range of Mucin Gene Expression in Normal Human Conjunctival Epithelium In Vivo . Invest. Ophthalmol. Vis. Sci. 2002;43(13):2464.
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Purpose: Our previous investigations proved that conjunctival impression cytology (CIC) is a valid, non-invasive technique that is useful to detect mRNA levels of ocular mucin genes in healthy individuals. This study was designed to perform the relative quantitation of MUC1, 2, 4, 5AC and 7 mucin genes in CIC samples from healthy males and females, younger and older of 60 years of age. Methods: Two polyethersulfone filters were applied to the superior bulbar conjunctiva of 38 healthy donors. Two age categories were considered: less that 60 years (7 males, 7 females), and 60 years or older (15 males, 9 females). Real time RT-PCR was performed using total RNA to examine the expression of MUC1, 2, 4, 5AC and 7. Their relative quantity was determined by SYBR® Green I dye detection. The housekeeping gene GAPDH was used to normalize the amount of expression levels. Data of mucin genes were analyzed using a logarithmic transformation to obtain a distribution closer to the normal distribution. Results: All mucin genes were detected. Transcripts for MUC2 were the lowest in all cases. Transcripts for MUC7 were the highest for all samples except in males younger of 60 years of age, where levels of MUC4 were the highest. There was not significant differences in any of the mucin genes expression between males and females. No differences existed between males younger of 60 years compared to males older of 60 years of age. However, MUC 1 and MUC7 levels were significantly lower in females of less that 60 years of age compared to females older than 60 years. No significant differences were found between males and females older than 60 years. But in the group of volunteers younger of 60 years of age, MUC4 expression was higher in males and MUC5AC was higher in females. Conclusion: We have defined the normal levels of ocular mucin genes expression in CIC samples from healthy males and females, and in two different groups of age. These normal values can be used to be further compared with those from pathological samples.
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