title: PKCγ, role in lens differentiation and gap junction coupling.
abstract: OBJECTIVE
  To determine the role of PKCγ in the regulation of gap junction coupling in the normal lens, we have compared the properties of coupling in lenses from wild type (WT) and PKC-γ knockout (KO) mice.
  
  
  METHODS
  Western blotting, confocal immunofluorescence microscopy, immunoprecipitation, RT-PCR and quantitative real time PCR were used to study gap junction protein and message expression; gap junction coupling conductance and pH gating were measured in intact lenses using impedance studies.
  
  
  RESULTS
  There were no gross differences in size, clarity, or expression of full-length Cx46 or Cx50 in lenses from WT and PKCγ KO mice. However, total Cx43 protein expression was ~150% higher in the KO lenses. In WT lenses, Cx43 was found only in epithelial cells whereas in KO lenses, its expression continued into the fiber cells. Gap junction coupling conductance in the differentiating fibers (DF) of PKCγ KO lenses was 34% larger than that of WT. In the mature fiber (MF), the effect was much larger with the KO lenses having an 82% increase in coupling over WT. pH gating of the DF fibers was not altered by the absence of PKCγ.
  
  
  CONCLUSIONS
  PKCγ has a major role in the regulation of gap junction expression and coupling in the normal lens.
authors: Department of Biochemistry, Kansas State University, Manhattan, Kansas 66506, USA.
authors: Satyabrata
authors: S
authors: Das
authors: Huan
authors: H
authors: Wang
authors: Samuel A
authors: SA
authors: Molina
authors: Francisco J
authors: FJ
authors: Martinez-Wittinghan
authors: Snehalata
authors: S
authors: Jena
authors: Leonie K
authors: LK
authors: Bossmann
authors: Kendra A
authors: KA
authors: Miller
authors: Richard T
authors: RT
authors: Mathias
authors: Dolores J
authors: DJ
authors: Takemoto