A Thy1-CFP DBA/2J mouse line with cyan fluorescent protein expression in retinal ganglion cells.
A Thy1-CFP DBA/2J mouse line with cyan fluorescent protein expression in retinal ganglion cells.
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DOI：
10.1017/s095252380999023x
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发表时间：
2009-11
影响因子：
1.9
通讯作者：
Brecha, Nicholas C.
中科院分区：
文献类型：
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作者：
Raymond, Iona D.;Pool, Angela L.;Vila, Alejandro;Brecha, Nicholas C.
开发了神经节细胞中表达青色荧光蛋白 (CFP) 报告基因的 DBA/2J (D2) 转基因小鼠系，用于分析进行性青光眼期间的神经节细胞。 Thy1-CFP D2 (CFP-D2) 系是通过同源繁殖产生色素性青光眼的 D2 系和在神经节细胞中表达 CFP 的 Thy1-CFP 系而创建的。 CFP-D2 后代的微卫星标记分析证实了 D2 isa 和 ipd 基因座的遗传包含。这些位点内的特定突变导致 D2 小鼠黑素体蛋白功能失调和青光眼表型。聚合酶链式反应分析证实包含 Thy1-CFP 转基因。 CFP荧光神经节细胞直径6-20μm，分布在所有视网膜区域，CFP突起遍布内丛状层，CFP荧光轴突位于纤维层和视神经头。使用针对神经节细胞标记物 NF-L、NeuN、Brn3a 和 SMI32 的抗体进行免疫组织化学来确认神经节细胞中的 CFP 表达。使用针对无长突细胞标记物 HPC-1 和 ChAT 的抗体进行免疫组织化学，以确认胆碱能无长突细胞中 CFP 的弱表达。 CFP-D2 小鼠出现青光眼表型，包括虹膜疾病、神经节细胞损失、纤维层磨损和眼内压升高。开发了具有表达 CFP 的神经节细胞的 CFP-D2 转基因系，其具有 (1) 主要为 D2 遗传背景，(2) 表达 CFP 的神经节细胞，以及 (3) 年龄相关的进行性青光眼。该细胞系对于研究青光眼进展过程中体内和体外神经节细胞及其轴突的实验研究具有价值。
A DBA/2J (D2) transgenic mouse line with cyan fluorescent protein (CFP) reporter expression in ganglion cells was developed for the analysis of ganglion cells during progressive glaucoma. The Thy1-CFP D2 (CFP-D2) line was created by congenically breeding the D2 line, which develops pigmentary glaucoma, and the Thy1-CFP line, which expresses CFP in ganglion cells. Microsatellite marker analysis of CFP-D2 progeny verified the genetic inclusion of the D2 isa and ipd loci. Specific mutations within these loci lead to dysfunctional melanosomal proteins and glaucomatous phenotype in D2 mice. Polymerase chain reaction analysis confirmed the inclusion of the Thy1-CFP transgene. CFP-fluorescent ganglion cells, 6–20 μm in diameter, were distributed in all retinal regions, CFP processes were throughout the inner plexiform layer, and CFP-fluorescent axons were in the fiber layer and optic nerve head. Immunohistochemistry with antibodies to ganglion cell markers NF-L, NeuN, Brn3a, and SMI32 was used to confirm CFP expression in ganglion cells. Immunohistochemistry with antibodies to amacrine cell markers HPC-1 and ChAT was used to confirm weak CFP expression in cholinergic amacrine cells. CFP-D2 mice developed a glaucomatous phenotype, including iris disease, ganglion cell loss, attrition of the fiber layer, and elevated intraocular pressure. A CFP-D2 transgenic line with CFP-expressing ganglion cells was developed, which has (1) a predominantly D2 genetic background, (2) CFP-expressing ganglion cells, and (3) age-related progressive glaucoma. This line will be of value for experimental studies investigating ganglion cells and their axons in vivo and in vitro during the progressive development of glaucoma.