Visual restoration and transplant connectivity in degenerate rats implanted with retinal progenitor sheets
Visual restoration and transplant connectivity in degenerate rats implanted with retinal progenitor sheets
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DOI：
10.1111/j.1460-9568.2010.07085.x
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发表时间：
2010-02-01
影响因子：
3.4
通讯作者：
Keirstead, H. S.
中科院分区：
文献类型：
--
作者：
Seiler, M. J.;Aramant, R. B.;Keirstead, H. S.
本研究的目的是确定视网膜祖层移植是否与宿主形成突触连接并恢复视力。将表达人胎盘碱性磷酸酶（human placental alkaline phosphatase, hPAP）的胚胎第19天大鼠胚胎分离的供体视网膜片移植到18只0.8 ~ 1.3月龄快速视网膜变性的S334ter-3大鼠的视网膜下间隙。受者在1.6-11.8个月大时被杀死。冷冻切片共聚焦免疫组织化学分析供体细胞标记hPAP和突触标记物。对振动体切片进行hPAP染色，并进行电镜处理。用电生理方法记录了12只5.3 ~ 11.8月龄大鼠上丘的视觉反应。所有记录的移植大鼠在视网膜移植位置对应的SC中恢复或保留了视觉反应，阈值在-2.8和-3.4 log cd/m之间(2)。在没有移植的年龄匹配的S334ter-3大鼠中，或在假手术的大鼠中，没有发现这种反应。通过光镜和电子显微镜在宿主体内鉴定供体细胞和过程。移植过程穿透宿主视网膜内部，尽管移植和宿主之间偶尔存在胶质屏障。在宿主内丛状层发现标记的神经元突起，并与未标记的细胞形成明显的突触，可能来自宿主。总之，移植物和宿主细胞之间的突触连接，以及大脑中相应位置的视觉反应，支持了视网膜层移植到啮齿动物视网膜变性模型后功能连接发展的假设。
The aim of this study was to determine whether retinal progenitor layer transplants form synaptic connections with the host and restore vision. Donor retinal sheets, isolated from embryonic day 19 rat fetuses expressing human placental alkaline phosphatase (hPAP), were transplanted to the subretinal space of 18 S334ter-3 rats with fast retinal degeneration at the age of 0.8-1.3 months. Recipients were killed at the age of 1.6-11.8 months. Frozen sections were analysed by confocal immunohistochemistry for the donor cell label hPAP and synaptic markers. Vibratome slices were stained for hPAP, and processed for electron microscopy. Visual responses were recorded by electrophysiology from the superior colliculus (SC) in 12 rats at the age of 5.3-11.8 months. All recorded transplanted rats had restored or preserved visual responses in the SC corresponding to the transplant location in the retina, with thresholds between -2.8 and -3.4 log cd/m(2). No such responses were found in age-matched S334ter-3 rats without transplants, or in those with sham surgery. Donor cells and processes were identified in the host by light and electron microscopy. Transplant processes penetrated the inner host retina in spite of occasional glial barriers between transplant and host. Labeled neuronal processes were found in the host inner plexiform layer, and formed apparent synapses with unlabeled cells, presumably of host origin. In conclusion, synaptic connections between graft and host cells, together with visual responses from corresponding locations in the brain, support the hypothesis that functional connections develop following transplantation of retinal layers into rodent models of retinal degeneration.