title: Orphan receptor GPR179 forms macromolecular complexes with components of metabotropic signaling cascade in retina ON-bipolar neurons.
abstract: OBJECTIVE
  In the mammalian retina, synaptic transmission between light-excited rod photoreceptors and downstream ON-bipolar neurons is indispensable for dim vision, and disruption of this process leads to congenital stationary night blindness in human patients. The ON-bipolar neurons use the metabotropic signaling cascade, initiated by the mGluR6 receptor, to generate depolarizing responses to light-induced changes in neurotransmitter glutamate release from the photoreceptor axonal terminals. Evidence for the identity of the components involved in transducing these signals is growing rapidly. Recently, the orphan receptor, GPR179, a member of the G protein-coupled receptor (GPCR) superfamily, has been shown to be indispensable for the synaptic responses of ON-bipolar cells. In our study, we investigated the interaction of GPR179 with principle components of the signal transduction cascade.
  
  
  METHODS
  We used immunoprecipitation and proximity ligation assays in transfected cells and native retinas to characterize the protein-protein interactions involving GPR179. The influence of cascade components on GPR179 localization was examined through immunohistochemical staining of the retinas from genetic mouse models.
  
  
  RESULTS
  We demonstrated that, in mouse retinas, GPR179 forms physical complexes with the main components of the metabotropic cascade, recruiting mGluR6, TRPM1, and the RGS proteins. Elimination of mGluR6 or RGS proteins, but not TRPM1, detrimentally affects postsynaptic targeting or GPR179 expression.
  
  
  CONCLUSIONS
  These observations suggest that the mGluR6 signaling cascade is scaffolded as a macromolecular complex in which the interactions between the components ensure the optimal spatiotemporal characteristics of signal transduction.
authors.affiliation: Department of Neuroscience, The Scripps Research Institute, Jupiter, Florida.
authors.forename: Cesare
authors.initials: C
authors.lastname: Orlandi
authors.forename: Yan
authors.initials: Y
authors.lastname: Cao
authors.forename: Kirill A
authors.initials: KA
authors.lastname: Martemyanov
mesh_terms: D000818:Animals; D015153:Blotting, Western; D005786:Gene Expression Regulation; D007150:Immunohistochemistry; D055537:Light Signal Transduction / Q000502:physiology*; D046911:Macromolecular Substances; D051379:Mice; D018345:Mice, Knockout; D020710:RGS Proteins / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism / Q000502:physiology*; D018094:Receptors, Metabotropic Glutamate / Q000378:metabolism; D051245:Retinal Bipolar Cells / Q000502:physiology*
chemical_list: C575060:GPR179 protein, mouse; D046911:Macromolecular Substances; D020710:RGS Proteins; D043562:Receptors, G-Protein-Coupled; D018094:Receptors, Metabotropic Glutamate; C113897:metabotropic glutamate receptor 6
keywords: ON-bipolar neurons; macromolecular complexes; orphan GPCR; protein targeting; retina