March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Identification of Retina-Specific Dynamin-1 Protein Complexes
Author Affiliations & Notes
  • Lindsey A. Ebke
    Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio
  • Gregory H. Grossman
    Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio
  • Gayle J. Pauer
    Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio
  • Craig D. Beight
    Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio
  • Geeng-Fu Jang
    Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio
  • John W. Crabb
    Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio
    Ophthalmology, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, Ohio
  • Stephanie A. Hagstrom
    Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio
    Ophthalmology, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, Ohio
  • Footnotes
    Commercial Relationships  Lindsey A. Ebke, None; Gregory H. Grossman, None; Gayle J. Pauer, None; Craig D. Beight, None; Geeng-Fu Jang, None; John W. Crabb, None; Stephanie A. Hagstrom, None
  • Footnotes
    Support  NIH Grant EY16072, Foundation Fighting Blindness, and Research to Prevent Blindness
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 2592. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Lindsey A. Ebke, Gregory H. Grossman, Gayle J. Pauer, Craig D. Beight, Geeng-Fu Jang, John W. Crabb, Stephanie A. Hagstrom; Identification of Retina-Specific Dynamin-1 Protein Complexes. Invest. Ophthalmol. Vis. Sci. 2012;53(14):2592.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: : Dynamin-1 is a highly-expressed neuronal-specific GTPase implicated in endocytosis and vesicle movement at presynaptic terminals of the brain; however, its role in the retina is unknown. We previously identified a retinal-specific isoform of Dynamin-1 that localizes to photoreceptor synaptic terminals and binds Tulp1, a photoreceptor-specific protein thought to be involved in vesicle movement. To further investigate this protein complex, we sought to identify additional Dynamin-1-binding partners in retinal tissue.

Methods: : Immunoprecipitation and Western blot experiments were performed with bovine and mouse retinal homogenates using a Dynamin-1 antibody raised against the epitope exclusive to the retinal-specific isoform. Immunoprecipitation products were separated by SDS-PAGE, protein bands were excised and in-gel digested with trypsin and identified by LC MS/MS. Identified proteins were localized in mouse retinal sections by immunohistochemistry.

Results: : We identified several canonical endocytic proteins by immunoprecipitation, including alpha-tubulin and clathrin. Most notably, we identified Dynamin-3, a protein thought to mediate vesicle cycling, in both bovine and murine retinal homogenates. Western blot analysis indicates that Dynamin-3 is expressed in the retina, and immunohistochemistry shows that it co-localizes with Dynamin-1 and Tulp1 in photoreceptor synaptic terminals. Reciprocal immunoprecipitation, immunohistochemistry and proximity ligand analyses are in progress to verify and further evaluate these interactions.

Conclusions: : Our data indicate that Dynamin-1 is likely involved in the endocytic pathway of the retina. Dynamin-3 is expressed in photoreceptor cells and may interact with the Dynamin-1/Tulp1 complex in the movement of vesicles in synaptic terminals. Further investigation into the nature of the interactions of these proteins will be carried out.

Keywords: photoreceptors • synapse • protein structure/function 
×
×

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.

×