June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Piezo1 channels highly expressed in human RPE
Author Affiliations & Notes
  • Qin Wan
    National Eye Institute, Bethesda, Maryland, United States
  • Joys Annika David
    National Eye Institute, Bethesda, Maryland, United States
  • Congxiao Zhang
    National Eye Institute, Bethesda, Maryland, United States
  • Tea Soon Park
    National Eye Institute, Bethesda, Maryland, United States
  • Mitra Farnoodian
    National Eye Institute, Bethesda, Maryland, United States
  • Eric Nguyen
    National Eye Institute, Bethesda, Maryland, United States
  • Jair Montford
    National Eye Institute, Bethesda, Maryland, United States
  • Colby Lewallen
    National Eye Institute, Bethesda, Maryland, United States
  • Aman George
    National Eye Institute, Bethesda, Maryland, United States
  • Kiyoharu Miyagishima
    National Eye Institute, Bethesda, Maryland, United States
  • David McGaughey
    National Eye Institute, Bethesda, Maryland, United States
  • Davide Ortolan
    National Eye Institute, Bethesda, Maryland, United States
  • Devika Abhijit Bose
    National Eye Institute, Bethesda, Maryland, United States
  • Arvydas Maminishkis
    National Eye Institute, Bethesda, Maryland, United States
  • Ruchi Sharma
    National Eye Institute, Bethesda, Maryland, United States
  • Kapil Bharti
    National Eye Institute, Bethesda, Maryland, United States
  • Footnotes
    Commercial Relationships   Qin Wan None; Joys Annika David None; Congxiao Zhang None; Tea Soon Park None; Mitra Farnoodian None; Eric Nguyen None; Jair Montford None; Colby Lewallen None; Aman George None; Kiyoharu Miyagishima None; David McGaughey None; Davide Ortolan None; Devika Bose None; Arvydas Maminishkis None; Ruchi Sharma None; Kapil Bharti None
  • Footnotes
    Support  NEI intramural grant support
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 2982. doi:
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      Qin Wan, Joys Annika David, Congxiao Zhang, Tea Soon Park, Mitra Farnoodian, Eric Nguyen, Jair Montford, Colby Lewallen, Aman George, Kiyoharu Miyagishima, David McGaughey, Davide Ortolan, Devika Abhijit Bose, Arvydas Maminishkis, Ruchi Sharma, Kapil Bharti; Piezo1 channels highly expressed in human RPE. Invest. Ophthalmol. Vis. Sci. 2023;64(8):2982.

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

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Abstract

Purpose : Piezo1 is mechanically activated cation channel and plays critical roles in a variety of physiological and pathological processes involving mechanosensation to shear stress or mechanical stretch. Retinal Pigmented Epithelium (RPE) monolayer faces the fluid-filled subretinal space, the tight control of the volume and composition of which is essential for maintaining the adjacent photoreceptor health. Excessive fluid accumulation in macular edema causes severe damage to RPE and leads to photoreceptor death. Our study investigates the expression and electrical properties of Piezo1 channel in RPE and its role in RPE development and function.

Methods : Human iPSC derived RPE (iPSC-RPE) were grown on semi-permeable transwells to generate a confluent monolayer. RNAseq, immunostaining, Western blot, qPCR, in combination with intracellular microelectrode recording were used to evaluate Piezo1 expression and activity.

Results : RNAseq demonstrate Piezo1 is highly expressed at comparable levels in human fetal, adult, and iPSC-RPE. Immunostaining revealed its localization mainly on the apical membrane of iPSC-RPE, with mild expression in the cytosol. Western blot confirmed the MW around 286kDa. Intracellular recordings demonstrated that apical application of Piezo1 activator Yoda1 produced a substantial drop in transepithelial potential (TEP) and resistance (TER), accompanied by an apical membrane depolarization and a decreased Ra/Rb ratio, suggesting the activation of an apical Ca2+/Na+ conductance. In contrast, basal application only induced subtle electrical response. qPCR analysis revealed that Piezo1 mRNA level shows dynamic changes during RPE development with a peak on two weeks post seeding on transwells. Immunostaining shows that Piezo1 was mainly localized in the cytosol till day14, and then translocated to the apical membrane as RPE matures. Chronic activation of Piezo1 with Yoda1 suppressed monolayer TER, and a large portion of Piezo1 was translocated from RPE apical membrane to the cytosol, accompanied by the changed distribution of Ezrin and F-actin filaments. The treatment with Piezo1 channel blocker GsMTx4 had no effect on TER but induced profound changes in the RPE apical processes.

Conclusions : Piezo1 channels are highly expressed in human RPE, mainly localized on RPE apical membrane. Early expression of Piezo1 in RPE development and its interaction with Ezrin and actin filaments implicate its possible role in RPE apical process development.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

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