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Yoshitsugu Saishin, Ryoki Ishikawa, Shinya Ugawa, Wei Guo, Takashi Ueda, Hiroyuki Morimura, Kazuhiro Kohama, Hideo Shimizu, Yasuo Tano, Shoichi Shimada; Retinal Fascin: Functional Nature, Subcellular Distribution, and Chromosomal Localization. Invest. Ophthalmol. Vis. Sci. 2000;41(8):2087-2095.
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purpose. To investigate the functional properties, subcellular localization, and
chromosomal location of retinal fascin.
methods. Recombinant retinal fascin protein was prepared by using a
baculovirus–insect expression system. Actin-binding and -bundling
assays were performed with chick actin purified from skeletal muscle.
Western blot analysis and immunohistochemistry were performed with a
polyclonal antibody raised against bovine retinal fascin. A human
retinal cDNA library was screened with an expressed sequence tag cDNA
fragment. Chromosomal location was determined with fluorescent in situ
results. The actin-binding and actin-bundling activities of retinal fascin were
demonstrated by high- and low-speed centrifugation assays. Formation of
filamentous (F)-actin bundles by retinal fascin in vitro was also
morphologically confirmed by fluorescence microscopy and electron
microscopy. Immunohistochemical analysis revealed that retinal fascin
protein was localized specifically in the outer and inner segments of
the photoreceptor cells in the retina. Two splicing variants of human retinal fascin cDNA were also located. One clone encoded 492
amino acids, and the other encoded 516 amino acids. The gene encoding
retinal fascin was localized to human chromosome 17, region q24–25.
conclusions. These results suggest that retinal fascin may play a role in formation
of unique morphologic structures of the photoreceptor cells and is a
candidate gene for retinal degenerative
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