Molecular studies into cell biological role of Copine-4 in Retinal Ganglion Cells

PLoS One. 2021 Nov 30;16(11):e0255860. doi: 10.1371/journal.pone.0255860. eCollection 2021.

Abstract

The molecular mechanisms underlying morphological diversity in retinal cell types are poorly understood. We have previously reported that several members of the Copine family of Ca-dependent membrane adaptors are expressed in Retinal Ganglion Cells and transcriptionally regulated by Brn3 transcription factors. Several Copines are enriched in the retina and their over-expression leads to morphological changes -formation of elongated processes-, reminiscent of neurites, in HEK293 cells. However, the role of Copines in the retina is largely unknown. We now investigate Cpne4, a Copine whose expression is restricted to Retinal Ganglion Cells. Over-expression of Cpne4 in RGCs in vivo led to formation of large varicosities on the dendrites but did not otherwise visibly affect dendrite or axon formation. Protein interactions studies using yeast two hybrid analysis from whole retina cDNA revealed two Cpne4 interacting proteins-Host Cell Factor 1 and Morn2. Mass Spectrometry analysis of retina lysate pulled down using Cpne4 or its vonWillebrand A domain showed 207 interacting proteins. A Gene Ontology analysis of the discovered proteins suggests that Cpne4 is involved in several metabolic and signaling pathways in the retina.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axons / metabolism*
  • Calcium-Binding Proteins / genetics
  • Calcium-Binding Proteins / metabolism*
  • HEK293 Cells
  • Humans
  • Mice
  • Mice, Knockout
  • Neurites / metabolism
  • Retina / metabolism*
  • Retinal Ganglion Cells / metabolism*
  • Transfection

Substances

  • Calcium-Binding Proteins

Grants and funding

This study was funded by National Eye Institute (NIH) intramural funding support to Tudor C. Badea (EY000504), and Graeme Wistow (AEY000433). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.