title: Ablation of Kcnj10 expression in retinal explants revealed pivotal roles for Kcnj10 in the proliferation and development of Müller glia.
abstract: OBJECTIVE
  We previously found that Kcnj10, an inwardly-rectifying potassium channel, is a gene expressed in c-kit-positive retinal progenitor cells on P1. The shRNA-mediated screening of the functions of the genes for retinal development in retinal explant culture suggested a role for Kcnj10 in the differentiation of 23Müller glia. In the present study, we extended the work and focused on analyzing the role of Kcnj10 in retinal development.
  
  
  METHODS
  shRNA-mediated downregulation of Kcnj10 in retinal explants and the in vivo mouse retina at the P1 stage was performed. Differentiation and proliferation of the retina were examined with immunohistochemistry. The effect of barium (Ba(2+)) treatment, which inhibits potassium currents by blocking potassium channels, on retinal development was examined.
  
  
  RESULTS
  When Kcnj10 was downregulated at E18, cellular proliferation and morphological differentiation were perturbed; in particular, a decreased number of Müller glial cells with abnormal morphological maturation was observed. The overexpression of Kcnj10 in retinal progenitors did not result in gross abnormality during retinal development, but rescued the abnormal differentiation induced with sh-Kcnj10. The presence of Ba(2+) in the retinal explant medium led to a phenotype similar to that seen with sh-Kcnj10. Ba(2+) exerts an effect mainly during late retinal development, and sh-Kcnj10 in the P1 retina affected Müller glia maturation, suggesting that Kcnj10 plays a pivotal role in the maturation of retinal cell subsets. A previous study of Kcnj10-knockout mice showed no obvious abnormality in retinal differentiation, especially of Müller glia. We examined the effects of the downregulation of Kcnj10 with in vivo electroporation of sh-Kcnj10 in the P1 retina. Retinal differentiation was perturbed, as seen following the in vitro downregulation of Kcnj10, suggesting that compensatory gene expression and/or signaling occurred in the Kcnj10-knockout mice in the retina, leading to normal eye development.
  
  
  CONCLUSIONS
  Kcnj10 plays a role in Müller glia maturation during retinal development probably through ionic channel activities.
authors: Division of Molecular and Developmental Biology, Institute of Medical Science, University of Tokyo, Tokyo, Japan ; Department of Ophthalmology, Juntendo University School of Medicine, Tokyo, Japan.
authors: Eisuke
authors: E
authors: Arai
authors: Division of Molecular and Developmental Biology, Institute of Medical Science, University of Tokyo, Tokyo, Japan.
authors: Yukihiro
authors: Y
authors: Baba
authors: Division of Molecular and Developmental Biology, Institute of Medical Science, University of Tokyo, Tokyo, Japan.
authors: Toshiro
authors: T
authors: Iwagawa
authors: Division of Molecular and Developmental Biology, Institute of Medical Science, University of Tokyo, Tokyo, Japan.
authors: Hiroshi
authors: H
authors: Kuribayashi
authors: Department of Ophthalmology, Juntendo University School of Medicine, Tokyo, Japan.
authors: Yujin
authors: Y
authors: Mochizuki
authors: Department of Ophthalmology, Juntendo University School of Medicine, Tokyo, Japan.
authors: Akira
authors: A
authors: Murakami
authors: Division of Molecular and Developmental Biology, Institute of Medical Science, University of Tokyo, Tokyo, Japan.
authors: Sumiko
authors: S
authors: Watanabe
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D017609:Barium Compounds / Q000494:pharmacology; D002452:Cell Count; D002454:Cell Differentiation; D019070:Cell Lineage / Q000235:genetics; D049109:Cell Proliferation; D002712:Chlorides / Q000494:pharmacology; D004622:Embryo, Mammalian; D063928:Ependymoglial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D051379:Mice; D008813:Mice, Inbred ICR; D018345:Mice, Knockout; D009024:Morphogenesis / Q000235:genetics*; D024661:Potassium Channels, Inwardly Rectifying / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D012160:Retina / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D015398:Signal Transduction; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D046509:Tissue Culture Techniques; D000098854:Kcnj10 Channel
chemical_list: D017609:Barium Compounds; D002712:Chlorides; D000098854:Kcnj10 Channel; D024661:Potassium Channels, Inwardly Rectifying; D034741:RNA, Small Interfering; C024986:barium chloride