Prolonged survival of the phosphorylated form of rhodopsin during dark adaptation of Royal College Surgeons rat
Prolonged survival of the phosphorylated form of rhodopsin during dark adaptation of Royal College Surgeons rat
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DOI：
10.1016/s0014-5793(03)00908-6
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发表时间：
2003-09-11
期刊：
影响因子：
3.5
通讯作者：
Nakazawa, M
中科院分区：
文献类型：
--
作者：
Ohguro, H;Ohguro, I;Nakazawa, M
为研究视紫红质（Rho）在皇家外科医学院（RCS）大鼠视网膜中的磷酸化和去磷酸化作用，用磷酸化多肽免疫家兔制备了针对Rho体内主要磷酸化位点334 Ser和338 Ser的特异性抗体。酶联免疫吸附试验表明，所产生的抗体仅识别磷酸化的334 Ser或338 Ser位点。在免疫荧光标记中，两种抗体识别来自Sprague-Dawley（SD）、Brown-Norway（BN）和RCS大鼠的光适应视网膜中的光感受器外节。在暗适应过程中，对磷酸化的338 Ser和334 Ser网站的免疫反应性在几个小时内（0.2-2小时）在SD和BN大鼠视网膜减少。然而，在RCS大鼠视网膜中，朝向磷酸化的338 Ser和334 Ser位点的那些在4至7天内减少。体外研究表明RCS视网膜中Rho磷酸化和去磷酸化反应的水平降低。然而，去磷酸化反应比磷酸化反应受到更大的影响。磷酸化形式的Rho极长的存活可能有助于RCS大鼠视网膜变性中光转导过程的持续失调。(C)2003年由Elsevier B. V.代表欧洲生物化学学会联合会出版。
To study rhodopsin (Rho) phosphorylation and dephosphorylation in Royal College of Surgeons (RCS) rat retina, specific antibodies toward major Rho phosphorylation sites in vivo, 334Ser or 338Ser, were prepared by immunization of authentic phosphorylated peptides in rabbit. Enzyme-linked immunosorbent assay identified that the raised antibodies exclusively recognized either the phosphorylated 334Ser or 338Ser site. In immunofluorescence labeling, both antibodies recognized photoreceptor outer segments in light-adapted retinas from Sprague-Dawley (SD), Brown-Norway (BN) and RCS rat. During dark adaptation, immunoreactivities toward phosphorylated 338Ser and 334Ser sites were diminished within several hours (0.2-2 h) in SD and BN rat retinas. However, those toward phosphorylated 338Ser and 334Ser sites were diminished within 4 to 7 days in RCS rat retinas. In vitro studies demonstrated decreased levels of both Rho phosphorylation and dephosphorylation reactions in RCS retinas. However, the dephosphorylation reaction was much more greatly affected than the phosphorylation reaction. Extremely prolonged survival of phosphorylated forms of Rho may contribute to persistent misregulation of phototransduction processes in retinal degeneration in RCS rat. (C) 2003 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.