Foveal Retinal Ganglion Cells Develop Altered Calcium Dynamics Weeks After Photoreceptor Ablation

IF 3.2 Q1 OPHTHALMOLOGY
Zhengyang Xu BSc , Karteek Kunala PhD , Peter Murphy BSc , Laura Patak BSc , Teresa Puthussery OD, PhD , Juliette McGregor PhD
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Abstract

Purpose

Physiological changes in retinal ganglion cells (RGCs) have been reported in rodent models of photoreceptor (PR) loss, but this has not been investigated in primates. By expressing both a calcium indicator (GCaMP6s) and an optogenetic actuator (ChrimsonR) in foveal RGCs of the macaque, we reactivated RGCs in vivo and assessed their response in the weeks and years after PR loss.

Design

We used an in vivo calcium imaging approach to record optogenetically evoked activity in deafferented RGCs in primate fovea. Cellular scale recordings were made longitudinally over a 10-week period after PR ablation and compared with responses from RGCs that had lost PR input >2 years prior.

Participants

Three eyes received PR ablation, the right eye of a male Macaca mulatta (M1), the left eye of a female Macaca fascicularis (M2), and the right eye of a male Macaca fascicularis (M3). Two animals were used for in vivo recording, 1 for histological assessment.

Methods

Cones were ablated with an ultrafast laser delivered through an adaptive optics scanning light ophthalmoscope (AOSLO). A 0.5 second pulse of 25 Hz 660 nm light optogenetically stimulated RGCs, and the resulting GCaMP fluorescence signal was recorded using an AOSLO. Measurements were repeated over 10 weeks immediately after PR ablation, at 2.3 years and in control RGCs.

Main Outcome Measures

The calcium rise time, decay constant, and sensitivity index of optogenetic-mediated RGC were derived from GCaMP fluorescence recordings from 221 RGCs (animal M1) and 218 RGCs (animal M2) in vivo.

Results

After PR ablation, the mean decay constant of the calcium response in RGCs decreased 1.5-fold (standard deviation 1.6 ± 0.5 seconds to 0.6 ± 0.3 seconds) over the 10-week observation period in subject 1 and 2.1-fold (standard deviation 2.5 ± 0.5 seconds to 1.2 ± 0.2 seconds) within 8 weeks in subject 2. Calcium rise time and sensitivity index were stable. Optogenetic reactivation remained possible 2.3 years after PR ablation.

Conclusions

Altered calcium dynamics developed in primate foveal RGCs in the weeks after PR ablation. The mean decay constant of optogenetic-mediated calcium responses decreased 1.5- to twofold. This is the first report of this phenomenon in primate retina and further work is required to understand the role these changes play in cell survival and activity.

Financial Disclosure(s)

Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.

眼窝视网膜神经节细胞在感光细胞消融数周后出现钙动力学改变
目的 据报道,在感光受体(PR)缺失的啮齿类动物模型中,视网膜神经节细胞(RGC)发生了生理变化,但在灵长类动物中尚未进行过研究。通过在猕猴的眼窝 RGCs 中表达钙指示剂(GCaMP6s)和光遗传致动器(ChrimsonR),我们在体内重新激活了 RGCs,并评估了它们在 PR 缺失后数周和数年内的反应。参与者三只眼睛接受了 PR 消融,分别是雄性猕猴的右眼(M1)、雌性猕猴的左眼(M2)和雄性猕猴的右眼(M3)。两只动物用于体内记录,一只用于组织学评估。方法通过自适应光学扫描光眼底镜(AOSLO)发射超快激光消融瞳孔。0.5 秒的 25 Hz 660 nm 光脉冲光遗传刺激 RGC,并使用 AOSLO 记录由此产生的 GCaMP 荧光信号。主要结果测量从体内 221 个 RGC(动物 M1)和 218 个 RGC(动物 M2)的 GCaMP 荧光记录中得出光遗传介导的 RGC 的钙上升时间、衰减常数和灵敏度指数。结果PR消融后,受试者1的RGCs钙响应平均衰减常数在10周观察期内下降了1.5倍(标准偏差为1.6±0.5秒至0.6±0.3秒),受试者2在8周内下降了2.1倍(标准偏差为2.5±0.5秒至1.2±0.2秒)。钙上升时间和灵敏度指数保持稳定。结论在 PR 消融后的几周内,灵长类眼窝 RGC 的钙动力学发生了改变。光遗传介导的钙离子反应的平均衰减常数下降了 1.5 到 2 倍。这是首次报道灵长类视网膜中的这一现象,需要进一步的工作来了解这些变化在细胞存活和活动中发挥的作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Ophthalmology science
Ophthalmology science Ophthalmology
CiteScore
3.40
自引率
0.00%
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审稿时长
89 days
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