June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Use of Pentachloropseudilin to study the in vivo molecular function of the motor protein MYO1C for Opsin transport in Zebrafish Photoreceptors
Author Affiliations & Notes
  • Andrew Gruesen
    Department of Ophthalmology and Visual Neurosciences, University of Minnesota Twin Cities, Minneapolis, Minnesota, United States
  • Anjelynt Lor
    Department of Ophthalmology and Visual Neurosciences, University of Minnesota Twin Cities, Minneapolis, Minnesota, United States
  • Rakesh Radhakrishnan
    Department of Ophthalmology and Visual Neurosciences, University of Minnesota Twin Cities, Minneapolis, Minnesota, United States
  • Ahmed Sadah
    Department of Ophthalmology and Visual Neurosciences, University of Minnesota Twin Cities, Minneapolis, Minnesota, United States
  • Matthias Leung
    Department of Ophthalmology and Visual Neurosciences, University of Minnesota Twin Cities, Minneapolis, Minnesota, United States
  • Dorothy Li
    Department of Ophthalmology and Visual Neurosciences, University of Minnesota Twin Cities, Minneapolis, Minnesota, United States
  • Jeremy Steinman
    Department of Ophthalmology and Visual Neurosciences, University of Minnesota Twin Cities, Minneapolis, Minnesota, United States
  • Heidi Roehrich
    Department of Ophthalmology and Visual Neurosciences, University of Minnesota Twin Cities, Minneapolis, Minnesota, United States
  • Rene Martin
    Technische Universitat Dresden, Dresden, Sachsen, Germany
  • Altaf A. Kondkar
    Department of Ophthalmology, King Saud University College of Medicine, Riyadh, Saudi Arabia
  • Rafael Monsanto
    Department of Otolaryngology, University of Minnesota Twin Cities, Minneapolis, Minnesota, United States
  • Stephanie A Hagstrom
    Ophthalmic Research, Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio, United States
  • Beata Jastrzebska
    Department of Pharmacology, Case Western Reserve University, Cleveland, Ohio, United States
  • Hans-Joachim Knölker
    Technische Universitat Dresden, Dresden, Sachsen, Germany
  • Glenn P Lobo
    Department of Ophthalmology and Visual Neurosciences, University of Minnesota Twin Cities, Minneapolis, Minnesota, United States
    School of Medicine, Medical University of South Carolina, Charleston, South Carolina, United States
  • Footnotes
    Commercial Relationships   Andrew Gruesen None; Anjelynt Lor None; Rakesh Radhakrishnan None; Ahmed Sadah None; Matthias Leung None; Dorothy Li None; Jeremy Steinman None; Heidi Roehrich None; Rene Martin None; Altaf Kondkar None; Rafael Monsanto None; Stephanie Hagstrom None; Beata Jastrzebska None; Hans-Joachim Knölker None; Glenn Lobo None
  • Footnotes
    Support  NIH Grant EY025034, NIH Grant EY030889, NIH Grant 3R01EY030889-03S1
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 4472. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Andrew Gruesen, Anjelynt Lor, Rakesh Radhakrishnan, Ahmed Sadah, Matthias Leung, Dorothy Li, Jeremy Steinman, Heidi Roehrich, Rene Martin, Altaf A. Kondkar, Rafael Monsanto, Stephanie A Hagstrom, Beata Jastrzebska, Hans-Joachim Knölker, Glenn P Lobo; Use of Pentachloropseudilin to study the in vivo molecular function of the motor protein MYO1C for Opsin transport in Zebrafish Photoreceptors. Invest. Ophthalmol. Vis. Sci. 2023;64(8):4472.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : The mislocalization of opsins in the photoreceptors of the retina has been previously correlated to the development of retinal diseases such as retinitis pigmentosa and Usher’s Syndrome. However, the mechanisms of this mislocalization are largely unknown. Previously, inhibition of MYO1C via the Myo1c-KO mouse model has been shown to cause rhodopsin mislocalization. Here, we attempted to further explore the in vivo function of MYO1C for opsin trafficking using a zebrafish (Danio rerio) model.

Methods : MYO1C, an unconventional motor protein, was inhibited in vivo with pentachloropseudilin (PClP), a specific inhibitor of MYO1C. Wild-type (WT) zebrafish were exposed directly to PClP through application of a PClP aqueous solution into their habitat water. Control fish were instead exposed to a Dimethyl sulfoxide (DMSO) aqueous solution. All fish were euthanized just prior to official cardiac death and underwent immunohistological study. Images of stained retinal samples were obtained and analyzed for obvious signs of opsin mislocalization in photoreceptors.

Results : Retinal sections were stained using Red/Green anti-opsin to observe long and medium wavelength binding cone opsins, and ZPR3 for rhodopsin in outer segments (OS). Zebrafish exposed to PClP exhibited obvious mistrafficking of these rod and cone opsins, as fluorescent images revealed an accumulation of opsin in the inner segment (IS) and outer nuclear layer (ONL). Comparatively, control WT samples exposed to DMSO exhibit consistent distributions of opsin in the OS.

Conclusions : The observed mislocalization of opsins to the inner segment of the zebrafish retina supports the use of PClP as an effective inhibitor of MYO1C in the zebrafish model. The results showing such mislocalization further support MYO1C’s role in rhodopsin and cone opsin trafficking. Given that comparable pathology using both mouse and zebrafish models can be demonstrated, there are strong implications to suggest further investigation into the role of MYO1C opsin trafficking and ultimately, its role in inherited retinal diseases.

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

×
×

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.

×