Fujii S, Escaño M F, Ishibashi K, Fujii M, Sekiya Y, Yamamoto M, Saijoh K
Department of Ophthalmology, Kobe University School of Medicine, Japan.
Invest Ophthalmol Vis Sci. 2000 May;41(6):1533-41.
To identify genes that are highly expressed in form-deprived retina-retinal pigment epithelium-choroid tissues. Neuroendocrine-specific proteins were found to be highly expressed.
mRNAs enriched in retina-retinal pigment epithelium-choroid tissues from 3-, 7-, and 14-day form-deprived chick eyes were isolated by differential display technique with cDNA library screening. Neuroendocrine-specific protein A and C were cloned in control and form-deprived eyes. mRNA and protein levels, with respective regional localizations, were examined by Northern blot, Western blot, and immunohistochemical analyses, respectively.
The isolated clone included an insert with a sequence homologous to both chick neuroendocrine-specific proteins A and C. The increases in mRNA and protein levels were confirmed by Northern and Western blot analyses, respectively. Immunohistochemical localization of neuroendocrine-specific proteins A and C was detected in the layer of photoreceptor inner segments, presumably in the cone cells. Northern blot analysis using negative lenses showed that levels of neuroendocrine-specific protein A and C mRNAs were not altered using negative lenses.
The expression of both neuroendocrine-specific proteins A and C mRNAs in cone photoreceptor cells was upregulated within 14 days of form deprivation, but not in response to negative spectacle lenses. These data suggest that the increase in induction of neuroendocrine-specific proteins is not a secondary consequence of ocular elongation or myopic refraction. Induction of neuroendocrine-specific proteins in form-deprived eyes may be causally related to the development of myopia or may be an unrelated effect of form deprivation.
鉴定在形觉剥夺性视网膜-视网膜色素上皮-脉络膜组织中高表达的基因。发现神经内分泌特异性蛋白高表达。
采用差异显示技术结合cDNA文库筛选，从3日龄、7日龄和14日龄形觉剥夺的鸡眼中分离富集于视网膜-视网膜色素上皮-脉络膜组织的mRNA。在对照眼和形觉剥夺眼中克隆神经内分泌特异性蛋白A和C。分别通过Northern印迹、Western印迹和免疫组织化学分析检测mRNA和蛋白质水平及其各自的区域定位。
分离的克隆包含一个与鸡神经内分泌特异性蛋白A和C序列同源的插入片段。Northern印迹和Western印迹分析分别证实了mRNA和蛋白质水平的增加。在光感受器内节层（推测在视锥细胞中）检测到神经内分泌特异性蛋白A和C的免疫组织化学定位。使用负透镜的Northern印迹分析表明，使用负透镜时神经内分泌特异性蛋白A和C的mRNA水平未改变。
在形觉剥夺的14天内，视锥光感受器细胞中神经内分泌特异性蛋白A和C的mRNA表达均上调，但对负性眼镜片无反应。这些数据表明，神经内分泌特异性蛋白诱导的增加不是眼轴伸长或近视性屈光的继发结果。形觉剥夺眼中神经内分泌特异性蛋白的诱导可能与近视的发生有因果关系，或者可能是形觉剥夺的无关效应。
