title: Multipronged approach to identify and validate a novel upstream regulator of Sncg in mouse retinal ganglion cells.
abstract: Loss of retinal ganglion cells (RGCs) is one of the hallmarks of retinal neurodegenerative diseases, glaucoma being one of the most common. Mechanistic studies on RGCs are hindered by the lack of sufficient primary cells and consensus regarding their signature markers. Recently, γ-synuclein (SNCG) has been shown to be highly expressed in the somas and axons of RGCs. In various mouse models of glaucoma, downregulation of Sncg gene expression correlates with RGC loss. To investigate the role of Sncg in RGCs, we used a novel systems genetics approach to identify a gene that modulates Sncg expression, followed by confirmatory studies in both healthy and diseased retinae. We found that chromosome 1 harbors an expression quantitative trait locus that modulates Sncg expression in the mouse retina, and identified the prefoldin-2 (PFDN2) gene as the candidate upstream modulator of Sncg expression. Our immunohistochemical analyses revealed similar expression patterns in both mouse and human healthy retinae, with PFDN2 colocalizing with SNCG in RGCs and their axons. In contrast, in retinae from glaucoma subjects, SNCG levels were significantly reduced, although PFDN2 levels were maintained. Using a novel flow cytometry-based RGC isolation method, we obtained viable populations of murine RGCs. Knocking down Pfdn2 expression in primary murine RGCs significantly reduced Sncg expression, confirming that Pfdn2 regulates Sncg expression in murine RGCs. Gene Ontology analysis indicated shared mitochondrial function associated with Sncg and Pfdn2. These data solidify the relationship between Sncg and Pfdn2 in RGCs, and provide a novel mechanism for maintaining RGC health.
authors.affiliation: Department of Ophthalmology, The Hamilton Eye Institute, The University of Tennessee Health Science Center, Memphis, TN, USA.|Department of Anatomy and Neurobiology, The University of Tennessee Health Science Center, Memphis, TN, USA.
authors.forename: Sumana R
authors.initials: SR
authors.lastname: Chintalapudi
authors.affiliation: Department of Ophthalmology, The Hamilton Eye Institute, The University of Tennessee Health Science Center, Memphis, TN, USA.|Department of Anatomy and Neurobiology, The University of Tennessee Health Science Center, Memphis, TN, USA.|Department of Microbiology, Immunology and Biochemistry, The University of Tennessee Health Science Center, Memphis, TN, USA.
authors.forename: Vanessa M
authors.initials: VM
authors.lastname: Morales-Tirado
authors.affiliation: Department of Anatomy and Neurobiology, The University of Tennessee Health Science Center, Memphis, TN, USA.|Department of Genetics, Genomics and Informatics, The University of Tennessee Health Science Center, Memphis, TN, USA.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Williams
authors.affiliation: Department of Ophthalmology, The Hamilton Eye Institute, The University of Tennessee Health Science Center, Memphis, TN, USA.|Department of Anatomy and Neurobiology, The University of Tennessee Health Science Center, Memphis, TN, USA.
authors.forename: Monica M
authors.initials: MM
authors.lastname: Jablonski
mesh_terms: D000818:Animals; D005434:Flow Cytometry; D051379:Mice; D008810:Mice, Inbred C57BL; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000378:metabolism*; D051845:gamma-Synuclein / Q000235:genetics / Q000378:metabolism*
chemical_list: D018832:Molecular Chaperones; D051845:gamma-Synuclein; C112806:prefoldin
keywords: flow cytometry; prefoldin-2; primary retinal ganglion cells; small interfering RNA transfection; systems genetics; γ-synuclein