Expression of βA3/A1-crystallin in the developing and adult rat eye
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Crystallins are very abundant structural proteins of the lens and are also expressed in other tissues. We have previously reported a spontaneous mutation in the rat βA3/A1-crystallin gene, termed Nuc1, which has a novel, complex, ocular phenotype. The current study was undertaken to compare the expression pattern of this gene during eye development in wild type and Nuc1 rats by in situ hybridization (ISH) and immunohistochemistry (IHC). βA3/A1-crystallin expression was first detected in the eyes of both wild type and Nuc1 rats at embryonic (E) day 12.5 in the posterior portion of the lens vesicle, and remained limited to the lens fibers throughout fetal life. After birth, βA3/A1-crystallin expression was also detected in the neural retina (specifically in the astrocytes and ganglion cells) and in the retinal pigmented epithelium (RPE). This suggested that βA3/A1-crystallin is not only a structural protein of the lens, but has cellular function(s) in other ocular tissues. In summary, expression of βA3/A1-crystallin is controlled differentially in various eye tissues with lens being the site of greatest expression. Similar staining patterns, detected by ISH and IHC, in wild type and Nuc1 animals suggest that functional differences in the protein, rather than changes in mRNA/protein level of expression, likely account for developmental abnormalities in Nuc1.
KeywordsAstrocytes βA3/A1crystallin Eye development Ganglion cells Lens Retina In situ hybridization
This work was supported by National Institute of Health Grants EY018636 (DS), EY019037 (DS), EY019037-S (DS), EY012221 (MKD) and Research to Prevent Blindness (an unrestricted grant to The Wilmer Eye Institute). We thank Spring Valley Laboratories, Woodbine, MD, for raising the βA3/A1-crystallin antibody; Drs. Tomohiro Masuda and Zhiyong Yang for help with in situ protocols and Ms. Stacey Hose for technical support. We thank Dr. Morton F. Goldberg of the Wilmer Eye Institute, Baltimore, Maryland and Dr. Bhaja K. Padhi from Health Canada, Tunney’s Pasture, Ottawa, Canada for critical reading and discussion of this manuscript.
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