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From the 1 Department of Biology, West Virginia University, Morgantown, West Virginia; 2 The Jules Stein Eye Institute, University of California, Los Angeles Medical School; and 3 Laboratory of Molecular and Developmental Biology, National Eye Institute, Bethesda, Maryland.
PURPOSE. To evaluate the expression of ßB2-crystallin mRNA and protein in rat, bovine, and human nonlens and nonocular tissues.
METHODS. ßB2-crystallin mRNA levels were detected by RT-PCR. ßB2-crystallin protein was purified from rat and bovine tissues by FPLC chromatography. FPLC fractions were analyzed by immunoblotting. The identity of ßB2-crystallin protein, isolated from the retina, was confirmed by protein microsequencing.
RESULTS. ßB2-crystallin transcript was detected in rat brain, rat testis, and human retina by RT-PCR. ßB2-crystallin transcript was not found in rat lung, heart, ovary, spleen, thymus, kidney, and liver or in human brain and testis. ßB2-crystallin protein was partially purified from and its identity confirmed in rat brain, rat testis, and bovine retina. The bovine retinal protein was further confirmed to be authentic ßB2-crystallin by protein microsequencing.
CONCLUSIONS. These results establish that ßB2-crystallin mRNA and protein are expressed in tissues outside of the lens and outside of the eye including retina, brain, and testis. Extralenticular and extraocular expression of ßB2-crystallin, coupled with its participation in phosphorylation pathways, suggests that it has nonrefractive functions in these tissues.
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