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G. Wang, W. K. Scott, P. Whitehead, B. L. Court, K. M. Spencer, J. L. Kovach, S. G. Schwartz, A. Agarwal, J. L. Haines, M. A. Pericak-Vance; Localization of Arms2 mRNA and Protein in Human Retina. Invest. Ophthalmol. Vis. Sci. 2010;51(13):4116.
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Controversy remains as to the subcellular localization of ARMS2 protein in human retina and cultured cells. At least three distinct patterns of ARMS2 protein localization (mitochondrial, cytosolic or secreted) have been reported. We previously reported the cytosolic localization of ARMS2 protein in cultured ARPE-19 cells. To further test ARMS2 gene expression pattern in human retina and to validate ARMS2 protein subcellular targeting, we used ARMS2 specific probes and three lines of ARMS2 antibody (one commercially available and two custom-made products) to localize ARMS2 mRNA and protein in human retina.
Digoxigenin (DIG)-labeled ARMS2 cRNA probes were prepared by cloning the full-length human ARMS2 cDNA to the vector pSPT18 and subsequent in vitro transcription using either SP6 (antisense) or T7 (sense) RNA polymerases. Ethanol fixed and paraffin embedded eye sections were obtained from 5 post mortem eyes of 5 individuals without any eye diseases. Fluorescence in situ hybridization showed that ARMS2 mRNA was expressed at a higher level in retina while at a lower level in choriocapillaris. ARMS2 mRNA was widely distributed across all retina layers including photoreceptors. Consistently, immunohistochemistry also showed strong ARMS2 immunostaining in retina. Double immunostaining further confirmed that ARMS2 is not co-localized with either a mitochondrial marker (MTCO2) or an extracellular marker (TIMP3), but is largely a cytosolic protein in human retina.
ARMS2 gene is widely expressed in human retina and ARMS2 protein is a cytosolic protein, not mitochondrial or secreted protein.
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