AAV-mediated, optogenetic ablation of Müller Glia leads to structural and functional changes in the mouse retina.
AAV-mediated, optogenetic ablation of Müller Glia leads to structural and functional changes in the mouse retina.
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DOI：
10.1371/journal.pone.0076075
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Flannery JG
中科院分区：
文献类型：
--
作者：
Byrne LC;Khalid F;Lee T;Zin EA;Greenberg KP;Visel M;Schaffer DV;Flannery JG
米勒神经胶质细胞是视网膜中的主要神经胶质细胞，为神经元提供结构和代谢支持，对于视网膜的完整性至关重要。 Müller 细胞与许多视网膜退行性疾病密切相关，包括 2 型黄斑毛细血管扩张症，其中中央视力损伤可能与 Müller 胶质细胞的原发性缺陷有关。在这里，我们使用了一种工程化的 Müller 特异性 AAV 变体（称为 ShH10），将光诱导毒性蛋白 KillerRed 传递到小鼠视网膜中的 Müller 细胞。我们描述了这些细胞特异性消融对视觉功能和视网膜结构的结果。玻璃体内注射后获得ShH10-KillerRed表达，然后用绿光照射眼睛以诱导毒性。 KillerRed 的诱导导致 Müller 细胞损失，并伴随着 Müller 细胞标记物谷氨酰胺合成酶和细胞视黄醛结合蛋白的减少、视紫红质和视锥细胞视蛋白的减少以及神经胶质纤维酸性蛋白的上调。穆勒细胞的损失还导致视网膜紊乱，包括外核层和感光器内节和外节变薄。薄切片的高分辨率成像揭示了光感受器从 ONL 的移位、玫瑰花状结构的形成以及吞噬细胞的存在。此外，穆勒细胞消融导致视网膜血管面积和体积增加，以及形成迂曲血管和血管渗漏。电生理测量显示视网膜功能下降，明视和暗视视网膜电图振幅下降明显。这些结果表明，Müller 细胞的丢失可导致进行性视网膜退行性疾病，并表明 AAV 在 Müller 细胞中传递诱导毒性蛋白可能有助于创建视网膜营养不良的大型动物模型。
Müller glia, the primary glial cell in the retina, provide structural and metabolic support for neurons and are essential for retinal integrity. Müller cells are closely involved in many retinal degenerative diseases, including macular telangiectasia type 2, in which impairment of central vision may be linked to a primary defect in Müller glia. Here, we used an engineered, Müller-specific variant of AAV, called ShH10, to deliver a photo-inducibly toxic protein, KillerRed, to Müller cells in the mouse retina. We characterized the results of specific ablation of these cells on visual function and retinal structure. ShH10-KillerRed expression was obtained following intravitreal injection and eyes were then irradiated with green light to induce toxicity. Induction of KillerRed led to loss of Müller cells and a concomitant decrease of Müller cell markers glutamine synthetase and cellular retinaldehyde-binding protein, reduction of rhodopsin and cone opsin, and upregulation of glial fibrillary acidic protein. Loss of Müller cells also resulted in retinal disorganization, including thinning of the outer nuclear layer and the photoreceptor inner and outer segments. High resolution imaging of thin sections revealed displacement of photoreceptors from the ONL, formation of rosette-like structures and the presence of phagocytic cells. Furthermore, Müller cell ablation resulted in increased area and volume of retinal blood vessels, as well as the formation of tortuous blood vessels and vascular leakage. Electrophysiologic measures demonstrated reduced retinal function, evident in decreased photopic and scotopic electroretinogram amplitudes. These results show that loss of Müller cells can cause progressive retinal degenerative disease, and suggest that AAV delivery of an inducibly toxic protein in Müller cells may be useful to create large animal models of retinal dystrophies.
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DOI：
10.1523/jneurosci.2841-12.2012
发表时间：
2012-11-07
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Shen W;Fruttiger M;Zhu L;Chung SH;Barnett NL;Kirk JK;Lee S;Coorey NJ;Killingsworth M;Sherman LS;Gillies MC
通讯作者：
Gillies MC
影响因子：
7.7
作者：
Li, Yongjun;Hazarika, Surovi;Annex, Brian H.
通讯作者：
Annex, Brian H.
影响因子：
12.4
作者：
Petrs-Silva, Hilda;Dinculescu, Astra;Hauswirth, William W.
通讯作者：
Hauswirth, William W.
影响因子：
4.4
作者：
Otteson, Deborah C.;Phillips, M. Joseph
通讯作者：
Phillips, M. Joseph
影响因子：
7.7
作者：
Nishikiori, Nami;Osanai, Makoto;Sawada, Norimasa
通讯作者：
Sawada, Norimasa