June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Differences in primary cilia amongst retinal ganglion cell subtypes
Author Affiliations & Notes
  • Tia Kowal
    Ophthalmology, Stanford University School of Medicine, Stanford, California, United States
  • Onkar Dhande
    Ophthalmology, Stanford University School of Medicine, Stanford, California, United States
  • Ke Ning
    Ophthalmology, Stanford University School of Medicine, Stanford, California, United States
  • Biao Wang
    Ophthalmology, Stanford University School of Medicine, Stanford, California, United States
  • Qing Wang
    Ophthalmology, Stanford University School of Medicine, Stanford, California, United States
  • Wendy Liu
    Ophthalmology, Stanford University School of Medicine, Stanford, California, United States
  • Nicolas Berbari
    Biology, Indiana University Purdue University Indianapolis, Indianapolis, Indiana, United States
  • Yang Hu
    Ophthalmology, Stanford University School of Medicine, Stanford, California, United States
  • Yang Sun
    Ophthalmology, Stanford University School of Medicine, Stanford, California, United States
  • Footnotes
    Commercial Relationships   Tia Kowal None; Onkar Dhande None; Ke Ning None; Biao Wang None; Qing Wang None; Wendy Liu None; Nicolas Berbari None; Yang Hu None; Yang Sun None
  • Footnotes
    Support  NIH grant F32-EY032775-01
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 976 – F0373. doi:
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    • Get Citation

      Tia Kowal, Onkar Dhande, Ke Ning, Biao Wang, Qing Wang, Wendy Liu, Nicolas Berbari, Yang Hu, Yang Sun; Differences in primary cilia amongst retinal ganglion cell subtypes. Invest. Ophthalmol. Vis. Sci. 2022;63(7):976 – F0373.

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

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Abstract

Purpose : Primary cilia are sensory organelles that project from the surface of most differentiated cells. These microtubule-based structures coordinate several signaling pathways that are involved in cell proliferation, migration, and differentiation. It is known that primary cilia play critical roles in eye development and photoreceptor function; however, less is known about what role the primary cilia might play in mature retinal ganglion cells (RGCs). To begin to understand these organelles, we assessed the presence or absence of a cilia membrane marker Arl13b and a widely distributed neuronal cilia marker AC3 in different subtypes of mouse RGCs.

Methods : Immunofluorescent staining for primary cilia axoneme markers, Arl13b and AC3, and basal body marker centrin 3 was performed on retinas isolated from adult (P25-P35) transgenic mice in which fluorophores were specifically expressed in only one known subtype of RGC. Additional subtype protein markers were analyzed using immunostaining. Subtype marker specificity was confirmed by colocalized with the pan-RGC marker RBPMS, and ARL13b or AC3 signal was only considered a primary cilium if adjacent to centrin 3 signal. Statistical analyses were performed using Student’s t-test, and p<0.05 was considered statistically significant.

Results : The presence of prototypical cilia markers Arl13b and AC3 in defined subtypes of retinal ganglion cells was analyzed. These markers were not expressed equally amongst the RGCs. Primarily AC3 positive cilia were found on alpha-RGCs identified by protein markers osteopontin (53% AC3 – 17% Arl13b), calretinin (53% AC3 – 44% Arl13b) and SMI32 (47% AC3 - 27% Arl13b). Whereas directionally selective RGCs either CART positive or Trhr positive differentially localize Arl13b or AC3, respectively in their primary cilia. Intrinsically photosensitive RGCs differentially localize Arl13b and AC3 based on melanopsin expression. Cilia in antibody labeled melanopsin cells primarily had Arl13b (30%), whereas transgenic mouse melanopsin (opn4) labeled RGCs primarily have AC3 (54%) positive cilia.

Conclusions : Primary cilia may be differentially organized between the subtypes of retinal ganglion cells. Further analyses are required to better understand what implications this may have on RGC subtype specific function.

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

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