Long-Term Effect of Optic Nerve Axotomy on the Retinal Ganglion Cell Layer
Long-Term Effect of Optic Nerve Axotomy on the Retinal Ganglion Cell Layer
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DOI：
10.1167/iovs.15-17195
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发表时间：
2015-09-01
影响因子：
4.4
通讯作者：
Agudo-Barriuso, Marta
中科院分区：
文献类型：
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作者：
Nadal-Nicolas, Francisco M.;Sobrado-Calvo, Paloma;Agudo-Barriuso, Marta
目的。分析视神经损伤对视网膜神经节细胞（RGCs）、黑视素（+）RGCs原位移位及其余神经节细胞层（GCL）细胞的远期影响。在成年白化大鼠中，左视神经被压碎（ONC）或横断（ONT）。损伤视网膜和对侧视网膜在增加的生存期（长达15个月）进行分析。为了研究所有GCL细胞和RGCs，用Brn3a和黑视素对视网膜进行免疫检测，以鉴定一般RGC群体（Brn3a(+)）和m(+)RGCs，并用4'，6-二氨基-2-苯基吲哚（DAPI）进行反染色。并分析了Brn3a(+)RGCs和m(+)RGCs向内核层移位的情况。在其他视网膜中，GCL中的胶质细胞被胶质纤维酸性蛋白（GFAP）或Iba1鉴定，在一些视网膜中，免疫检测到Brn3a、calretinin和γ -突触核蛋白。在RGC和m(+)RGC种群中，正位和移位RGC的行为相似。这两种病变都会导致RGCs呈指数级减少（ONC或ONT术后6个月生存率为4%-1%），但不会导致m(+)RGCs减少，其数量在1 - 15个月保持稳定（占初始人群的34%-44%）。γ -synuclein在RGCs和移位无突细胞（dACs）中表达，这使我们确认axtomy不影响后者，并确定在组成GCL的约217,406个细胞（不包括内皮细胞）中，10%是胶质细胞，50%是dACs，其余40%是RGCs。在GCL中，切除后只有RGCs丢失，黑视素（+）RGCs与其他RGCs在丢失过程和存活率上存在重要差异。
PURPOSE. To analyze the long-term effect of optic nerve injury on retinal ganglion cells (RGCs) and melanopsin(+)RGCs orthotopic and displaced, and on the rest of the ganglion cell layer (GCL) cells.METHODS. In adult albino rats, the left optic nerve was crushed (ONC) or transected (ONT). Injured and contralateral retinas were analyzed at increasing survival intervals (up to 15 months). To study all GCL cells and RGCs, retinas were immunodetected with Brn3a and melanopsin to identify the general RGC population (Brn3a(+)) and m(+)RGCs, and counter-stained with 4',6-diamidino-2-phenylindole (DAPI). Brn3a(+)RGCs and m(+)RGCs displaced to the inner nuclear layer were analyzed as well. In additional retinas, glial cells in the GCL were identified with glial fibrillary acidic protein (GFAP) or Iba1, and in some retinas, Brn3a, calretinin, and gamma-synuclein were immunodetected.RESULTS. Orthotopic and displaced RGCs behave similarly within the RGC and m(+)RGC populations. Both lesions cause an exponential loss of RGCs (4%-1% survival at 6 months after ONC or ONT), but not of m(+)RGCs, whose number remains stable from 1 to 15 months (34%-44% of the initial population). gamma-synuclein is expressed by RGCs and displaced amacrine cells (dACs), allowing us to confirm that axotomy does not affect the latter, and to determine that out of the approximately 217,406 cells that compose the GCL (excluding endothelia), 10% are glial cells, 50% dACs, and the remaining 40% are RGCs.CONCLUSIONS. In the GCL, only RGCs are lost after axotomy, and there are important differences in the course of loss and rate of survival between melanopsin(+)RGCs and the rest of RGCs.