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S.G. Remington, J.M. Crow, J.D. Nelson; Secretoglobins of the Mouse Lacrimal Gland . Invest. Ophthalmol. Vis. Sci. 2004;45(13):3868.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose:We compared mRNA expression in male and female mouse lacrimal glands to identify differentially expressed proteins in tears. Methods:Poly(A)+ RNA from male and female Swiss Webster mouse lacrimal glands was isolated and reverse transcribed. Differential display analysis coupled with the polymerase chain reaction (PCR) was used to identify mRNAs that were expressed at different levels in male and female lacrimal glands. These PCR products were re–amplified, subcloned and sequenced. The sequences were compared with those in the NIH databases. Poly(A)+ RNAs from several mouse tissues (harderian, lacrimal, submandibular, sublingual and parotid glands, and liver) were electrophoresed in an agarose gel and transferred to nylon membrane. The blot was sequentially hybridized with unique 32P–labeled fragments of subcloned sequences. Actin was the positive control. Results:We identified four mouse secretoglobins, GenBank accession #'s AF008595, AF272844, AY370634 and AY370635. In an alignment of AF008595 with androgen binding protein–alpha (XM_133375) of mouse salivary gland, the 5' and 3' untranslated regions (and the signal sequence) were more conserved (88% identity) than the protein coding region (72% identity). Likewise in an alignment of AF272844, AY370634 and AY370635, the 5' and 3' untranslated regions (and the signal sequence) were more conserved (85% identity) than the protein coding region (65% identity). Among the tissues tested, sequence–specific labeled fragments of each secretoglobin clone hybridized only to the lacrimal gland RNA. Two of the secretoglobin clones (AF008595 and AY370634) hybridized equivalently to both male and female lacrimal gland RNA; two of the secretoglobin clones (AF272844 and AY370635) hybridized primarily to male mouse lacrimal gland RNA. Conclusions:Four mouse secretoglobin clones exhibited lacrimal gland–specific expression. Two of these were expressed primarily in male mouse lacrimal gland. Higher sequence variability within the protein coding regions of these secretoglobin mRNAs would be consistent with an immunological function.
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