June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
In Vivo Evaluation of Naïve and Diseased RGC Activities at Single-Cell Level
Author Affiliations & Notes
  • Liang Li
    Ophthalmology, Stanford University School of Medicine, Palo Alto, California, United States
  • Fang Fang
    Ophthalmology, Stanford University School of Medicine, Palo Alto, California, United States
    Department of Ophthalmology, The Second Xiangya Hospital, Central South University Xiangya School of Medicine, Changsha, Hunan, China
  • Xue Feng
    Ophthalmology, Stanford University School of Medicine, Palo Alto, California, United States
  • Shaobo Zhang
    Department of Ophthalmology, University of California San Francisco School of Medicine, San Francisco, California, United States
  • David Andrew Miller
    Department of Biomedical Engineering, Northwestern University, Evanston, Illinois, United States
  • Pei Zhuang
    Ophthalmology, Stanford University School of Medicine, Palo Alto, California, United States
  • Haoliang Huang
    Ophthalmology, Stanford University School of Medicine, Palo Alto, California, United States
  • Pingting Liu
    Ophthalmology, Stanford University School of Medicine, Palo Alto, California, United States
  • Junting Liu
    Ophthalmology, Stanford University School of Medicine, Palo Alto, California, United States
  • Nripun Sredar
    Ophthalmology, Stanford University School of Medicine, Palo Alto, California, United States
  • Liang Liu
    Ophthalmology, Stanford University School of Medicine, Palo Alto, California, United States
  • Yang Sun
    Ophthalmology, Stanford University School of Medicine, Palo Alto, California, United States
  • Xin Duan
    Department of Ophthalmology, University of California San Francisco School of Medicine, San Francisco, California, United States
  • Jeffrey L Goldberg
    Ophthalmology, Stanford University School of Medicine, Palo Alto, California, United States
  • Hao Zhang
    Department of Biomedical Engineering, Northwestern University, Evanston, Illinois, United States
  • Yang Hu
    Ophthalmology, Stanford University School of Medicine, Palo Alto, California, United States
  • Footnotes
    Commercial Relationships   Liang Li None; Fang Fang None; Xue Feng None; Shaobo Zhang None; David Miller None; Pei Zhuang None; Haoliang Huang None; Pingting Liu None; Junting Liu None; Nripun Sredar None; Liang Liu None; Yang Sun None; Xin Duan None; Jeffrey Goldberg None; Hao Zhang None; Yang Hu None
  • Footnotes
    Support  EY024932
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 1849. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Liang Li, Fang Fang, Xue Feng, Shaobo Zhang, David Andrew Miller, Pei Zhuang, Haoliang Huang, Pingting Liu, Junting Liu, Nripun Sredar, Liang Liu, Yang Sun, Xin Duan, Jeffrey L Goldberg, Hao Zhang, Yang Hu; In Vivo Evaluation of Naïve and Diseased RGC Activities at Single-Cell Level. Invest. Ophthalmol. Vis. Sci. 2022;63(7):1849.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose : The characteristic feature of optic nerve injury and glaucoma is the progressive degeneration of RGCs and their axons. However, the neuronal activities of RGCs and their changes in disease models have not been studied longitudinally in vivo. Here we demonstrate the feasibility of directly visualizing light-evoked RGCs activities by cSLO-mediated high throughput RGC Ca2+ imaging. The powerful population and single-cell analytic strategies we developed revealed, for the first time, distinct dynamic RGC activity changes in optic nerve crush(ONC) and glaucoma models.

Methods : Due to the superb optical accessibility of the retina, fluorescence-labeled RGCs can be imaged noninvasively in vivo by cSLO. We specifically delivered Ca2+ indicators, jGCaMP7s, to the mouse RGCs by AAV2 and mSncg promoter. Thus, we employed cSLO customized with UV stimulation and jGCaMP7s, to optically record light-evoked activities of about 1200 RGCs/retina simultaneously in naïve living mice, as well as longitudinal recording RGC activities at multiple time points in ONC and glaucoma mice.

Results : In naïve retinas, 47 RGC clusters were recognized by unsupervised algorithms, and then hierarchically re-grouped into 9 groups with distinct ON, OFF, and ON-OFF responses to UV stimulation. Consistent with distinct transcriptomic changes in trauma and glaucoma, population analysis of longitudinal RGC Ca2+ imaging reveals initial increase, but later rapid loss of ON-RGC activities in ONC, versus mild decreased but well-preserved ON- and OFF-RGC activities in glaucoma. More strikingly, single-cell Ca2+ imaging tracing uncovers unprecedented conversions of RGC activities, majority of ON-to-OFF conversion in trauma and OFF-to-ON conversion in glaucoma. Thus, not static as assumed, individual RGC’s light-evoked activity is rather vibrant with unique transformation patterns in different diseases.

Conclusions : Our results demonstrate the potential of in vivo RGC Ca2+ imaging as a reliable, sensitive, direct and noninvasive RGC functional measurement at single-cell level, but with high throughput capability. This proof-of-concept study lays out the foundation for in vivo RGC function classification and evaluation with more visual stimuli under normal, disease, and therapeutic conditions, at both population and single-cell levels, which is invaluable in understanding RGC pathophysiology and identifying functional biomarkers for diverse optic neuropathies

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×