Retinal transplants restore visual responses: trans-synaptic tracing from visually responsive sites labels transplant neurons
Retinal transplants restore visual responses: trans-synaptic tracing from visually responsive sites labels transplant neurons
复制标题
DOI：
10.1111/j.1460-9568.2008.06279.x
复制
发表时间：
2008-07-01
影响因子：
3.4
通讯作者：
Aramant, Robert B.
中科院分区：
文献类型：
--
作者：
Seiler, Magdalene J.;Thomas, Biju B.;Aramant, Robert B.
本研究旨在验证上级丘（SC）的视觉反应起源于胚胎视网膜移植物与退化的宿主视网膜之间的突触连接的假设。将表达人胎盘碱性磷酸酶的胚胎第19天大鼠视网膜的薄片移植到患有快速视网膜变性的3至4周龄S334 ter-line-3大鼠的视网膜下空间。几个月后，从SC中记录到了视觉反应。减毒伪狂犬病病毒特异性地在突触处的神经元之间转移（菌株PRV-152，表达绿色荧光蛋白（GFP）或BaBlu，表达大肠杆菌β-半乳糖苷酶）注射到SC的视觉响应位点。存活1-2天后，通过免疫组织化学结合不同的视网膜细胞标记物，如蛋白激酶C、恢复蛋白、钙-钙调蛋白依赖性蛋白激酶II和谷氨酰胺合成酶，检测视网膜中的病毒。移植的大鼠在视网膜中与移植物位置相对应的SC小区域的平均反应阈值为-3.1 log cd/m2。在注射到该SC区域后30小时，病毒追溯到覆盖移植物并靠近移植物的宿主神经节细胞。注射后2天，在宿主视网膜中发现广泛的病毒标记，并且移植物中的许多细胞也被标记。移植物中的病毒标记细胞被双重标记为神经元和神经胶质细胞标记物。这项研究提供了解剖学证据，胎儿视网膜移植和宿主视网膜之间的突触连接有助于SC中的视觉反应。
This study aimed to test the hypothesis that visual responses in the superior colliculus (SC) originate from synaptic connections between fetal retinal transplants and degenerating host retinas. Sheets of embryonic day 19 rat retina expressing human placental alkaline phosphatase were transplanted to the subretinal space of 3- to 4-week-old S334ter-line-3 rats with fast retinal degeneration. Several months later, visual responses were recorded from the SC. Attenuated pseudorabies virus that is specifically transferred between neurons at synapses (strains PRV-152, expressing green fluorescent protein (GFP) or BaBlu, expressing Escherichia coli beta-galactosidase) was injected into the visually responsive site of the SC. After survival times of 1-2 days, the virus was detected in the retina by immunohistochemistry in combination with different retinal cell markers, such as protein kinase C, recoverin, calcium-calmodulin-dependent protein kinase II and glutamine synthetase. Transplanted rats had a mean response threshold of -3.1 log cd/m(2) in a small area of the SC corresponding to the location of the graft in the retina. By 30 h after injection into this SC area, the virus traced back to host ganglion cells overlying the transplant and in close proximity to the transplant. By 2 days after injection, extensive virus label was found in the host retina and many cells in the transplant were also labeled. Virus-labeled cells in the transplant were double labeled for neuronal and glial cell markers. This study provides anatomical evidence that synaptic connections between fetal retinal transplants and host retinas contribute to the visual responses in the SC.