title: Kir4.1 and AQP4 associate with Dp71- and utrophin-DAPs complexes in specific and defined microdomains of Müller retinal glial cell membrane.
abstract: The dystrophin-associated proteins (DAPs) complex consisting of dystroglycan, syntrophin, dystrobrevin, and sarcoglycans in muscle cells is associated either with dystrophin or its homolog utrophin. In rat retina, a similar complex was found associated with dystrophin-Dp71 that serves as an anchor for the inwardly rectifying potassium channel Kir4.1 and the aqueous pore, aquaporin-4 (AQP4). Here, using immunofluorescence imaging of isolated retinal Müller glial cells and co-immunoprecipitation experiments performed on an enriched Müller glial cells end-feet fraction, we investigated the effect of Dp71 deletion on the composition, anchoring, and membrane localization of the DAPs-Kir4.1 and/or -AQP4 complex. Two distinct complexes were identified in the end-feet fraction associated either with Dp71 or with utrophin. Upon Dp71 deletion, the corresponding DAPs complex was disrupted and a compensating utrophin upregulation was observed, accompanied by diffuse overall staining of Kir4.1 along the Müller glial cells and redistribution of the K(+) conductance. Dp71 deficiency was also associated with a marked reduction of AQP4 and beta-dystroglycan expression. Furthermore, it was observed that the Dp71-DAPs dependent complex could be, at least partially, associated with a specific membrane fraction. These results demonstrate that Dp71 has a central role in the molecular scaffold responsible for anchoring AQP4 and Kir4.1 in Müller cell end-feet membranes. They also show that despite its close relationship to the dystrophin proteins and its correlated upregulation, utrophin is only partially compensating for the absence of Dp71 in Müller glial cells.
authors.affiliation: Université Pierre et Marie Curie-Paris6, UMR-S 592, Paris, France.
authors.forename: Patrice E
authors.initials: PE
authors.lastname: Fort
authors.forename: Abdoulaye
authors.initials: A
authors.lastname: Sene
authors.forename: Thomas
authors.initials: T
authors.lastname: Pannicke
authors.forename: Michel J
authors.initials: MJ
authors.lastname: Roux
authors.forename: Valerie
authors.initials: V
authors.lastname: Forster
authors.forename: Dominique
authors.initials: D
authors.lastname: Mornet
authors.forename: Uri
authors.initials: U
authors.lastname: Nudel
authors.forename: David
authors.initials: D
authors.lastname: Yaffe
authors.forename: Andreas
authors.initials: A
authors.lastname: Reichenbach
authors.forename: Jose A
authors.initials: JA
authors.lastname: Sahel
authors.forename: Alvaro
authors.initials: A
authors.lastname: Rendon
mesh_terms: D000818:Animals; D051401:Aquaporin 4 / Q000378:metabolism*; D016189:Dystrophin / Q000172:deficiency / Q000378:metabolism*; D049028:Dystrophin-Associated Protein Complex / Q000378:metabolism*; D047468:Immunoprecipitation / Q000379:methods; D021962:Membrane Microdomains / Q000378:metabolism*; D008564:Membrane Potentials / Q000502:physiology; D051379:Mice; D018345:Mice, Knockout; D008954:Models, Biological; D009457:Neuroglia / Q000166:cytology*; D018408:Patch-Clamp Techniques; D024661:Potassium Channels, Inwardly Rectifying / Q000378:metabolism*; D012160:Retina / Q000166:cytology*; D049411:Utrophin / Q000378:metabolism*; D000098854:Kcnj10 Channel
chemical_list: D051401:Aquaporin 4; D016189:Dystrophin; D049028:Dystrophin-Associated Protein Complex; D000098854:Kcnj10 Channel; D024661:Potassium Channels, Inwardly Rectifying; D049411:Utrophin; C085339:apo-dystrophin 1