Retinal ganglion cell death and optic nerve degeneration by genetic ablation in adult mice.
Retinal ganglion cell death and optic nerve degeneration by genetic ablation in adult mice.
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DOI：
10.1016/j.exer.2008.11.021
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发表时间：
2009-03
影响因子：
3.4
通讯作者：
Klein, William H.
中科院分区：
文献类型：
--
作者：
Cho, Jang-Hyeon;Mu, Xiuqian;Wang, Steven W.;Klein, William H.
关键词：
尽管问题很严重，但尚无有效的治疗方法可以预防影响人眼的疾病中视网膜神经节细胞（RGC）死亡和视神经变性的发生。目前可用于制定治疗策略的动物模型面临诱导 RGC 死亡所需的繁琐程序，或者依赖于诱导发育中视网膜缺陷而不是成人成熟视网膜缺陷的突变。我们的目标是开发一个强大的基因工程成年小鼠模型，用于基于基因消融的 RGC 损失和视神经变性。为了实现这一目标，我们利用了 Pou4f2 (Brn3b)，这是一种当 RGC 开始分化并在整个生命过程中表达时立即激活的基因。我们生成了成年小鼠，其基因组含有条件 Pou4f2 等位基因，其中含有 floxed-lacZ-stop-白喉毒素 A 盒和 CAGG-Cre-ER™ 转基因。在这个双基因模型中，Cre重组酶与修饰的雌激素核受体融合，其中雌激素结合结构域优先与雌激素激动剂他莫昔芬结合，而不是与内源性雌二醇结合。与雌激素结合结构域结合后，他莫昔芬会去抑制 Cre 重组酶，从而导致 floxed lacZ-stop DNA 序列的有效基因组删除和白喉毒素 A 的表达。通过腹腔注射给不同年龄的成年小鼠施用他莫昔芬，会导致 RGC 快速损失、反应性神经胶质增生、视神经在几个月内进行性退化以及视力障碍。也许更能反映人类疾病，RGC 的部分丧失是通过调节他莫昔芬治疗来实现的。尤其与人类视网膜病理中的 RGC 死亡和视神经变性相关，RGC 消融的视网膜保持了其结构完整性，而其他视网膜神经元及其在内、外丛状层中的连接似乎未受到 RGC 消融的影响。这些事件是在人类视网膜病理学中观察到的进行性视神经变性的标志，并证明了该模型用于开发干细胞疗法以用健康的 RGC 替代死亡的 RGC 的有效性。
Despite the magnitude of the problem, no effective treatments exist to prevent retinal ganglion cell (RGC) death and optic nerve degeneration from occurring in diseases affecting the human eye. Animal models currently available for developing treatment strategies suffer from cumbersome procedures required to induce RGC death or rely on mutations that induce defects in developing retinas rather than in mature retinas of adults. Our objective was to develop a robust genetically engineered adult mouse model for RGC loss and optic nerve degeneration based on genetic ablation. To achieve this, we took advantage of Pou4f2 (Brn3b), a gene activated immediately as RGCs begin to differentiate and expressed throughout life. We generated adult mice whose genomes harbored a conditional Pou4f2 allele containing a floxed-lacZ-stop-diphtheria toxin A cassette and a CAGG-Cre-ER™ transgene. In this bigenic model, Cre recombinase is fused to a modified estrogen nuclear receptor in which the estrogen-binding domain binds preferentially to the estrogen agonist tamoxifen rather than to endogenous estradiol. Upon binding to the estrogen-binding domain, tamoxifen derepresses Cre recombinase, leading to the efficient genomic deletion of the floxed lacZ-stop DNA sequence and expression of diphtheria toxin A. Tamoxifen administered to adult mice at different ages by intraperitoneal injection led to rapid RGC loss, reactive gliosis, progressive degradation of the optic nerve over a period of several months, and visual impairment. Perhaps more reflective of human disease, partial loss of RGCs was achieved by modulating the tamoxifen treatment. Especially relevant for RGC death and optic nerve degeneration in human retinal pathologies, RGC-ablated retinas maintained their structural integrity, and other retinal neurons and their connections in the inner and outer plexiform layers appeared unaffected by RGC ablation. These events are hallmarks of progressive optic nerve degeneration observed in human retinal pathologies and demonstrate the validity of this model for use in developing stem cell therapies for replacing dead RGCs with healthy ones.
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影响因子：
15.9
作者：
Harada, Takayuki;Harada, Chikako;Tanaka, Kohichi
通讯作者：
Tanaka, Kohichi
DOI：
10.1073/pnas.93.25.14955
发表时间：
1996-12-10
影响因子：
11.1
作者：
Porciatti, V;Pizzorusso, T;Maffei, L
通讯作者：
Maffei, L
影响因子：
2.7
作者：
Grieshammer, U;Lewandoski, M;Martin, GR
通讯作者：
Martin, GR
影响因子：
2.7
作者：
Gan, L;Wang, SW;Klein, WH
通讯作者：
Klein, WH
影响因子：
4.4
作者：
Kitaoka, Y;Kitaoka, Y;Lam, TT
通讯作者：
Lam, TT