Primary Cultures of Embryonic Chick Lens Cells as a Model System to Study Lens Gap Junctions and Fiber Cell Differentiation
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A major limitation in lens gap junction research has been the lack of experimentally tractable ex vivo systems to study the formation and regulation of fiber-type gap junctions. Although immortalized lens-derived cell lines are amenable to both gene transfection and siRNA-mediated knockdown, to our knowledge none are capable of undergoing appreciable epithelial-to-fiber differentiation. Lens central epithelial explants have the converse limitation. A key advance in the field was the development of a primary embryonic chick lens cell culture system by Drs. Sue Menko and Ross Johnson. Unlike central epithelial explants, these cultures also include cells from the peripheral (preequatorial and equatorial) epithelium, which is the most physiologically relevant population for the study of fiber-type gap junction formation. We have modified the Menko/Johnson system and refer to our cultures as dissociated cell-derived monolayer cultures (DCDMLs). We culture DCDMLs without serum to mimic the avascular lens environment and on laminin, the major matrix component of the lens capsule. Here, I review the features of the DCDML system and how we have used it to study lens gap junctions and fiber cell differentiation. Our results demonstrate the power of DCDMLs to generate new findings germane to the mammalian lens and how these cultures can be exploited to conduct experiments that would be impossible, prohibitively expensive and/or difficult to interpret using transgenic animals in vivo.
KeywordsGap junction Lens FGF Connexin
This work was supported by grant R01 2EY014622 from the National Eye Institute (to L.S.M.). I thank Ross Johnson for generously providing antibodies to lens proteins and for many interesting and helpful discussions over the years.
- Goldberg GS, Moreno AP, Bechberger JF, Hearn SS, Shivers RR, MacPhee DJ, Zhang YC, Naus CC (1996) Evidence that disruption of connexon particle arrangements in gap junction plaques is associated with inhibition of gap junctional communication by a glycyrrhetinic acid derivative. Exp Cell Res 222:48–53PubMedCrossRefGoogle Scholar
- Musil LS, Boswell BA (2010) Molecular mechanism of cross-talk between FGFs and BMPs in lens cell differentiation. Invest Ophthalmol Vis Sci 51; abstract 1213Google Scholar
- Reneker LW (2008) ERK1/2 signaling is required for epithelial-to-fiber differentiation in lens development. Invest Ophthalmol Vis Sci 49; abstract 1135Google Scholar
- Reneker LW, Robinson ML, Ogata M, Sanjo H, Pages G (2007) The role of ERK mitogen-activated protein kinase (MAPK) in lens development. Invest Ophthalmol Vis Sci 48; abstract 1119Google Scholar
- Xia CH, Cheng C, Huang Q, Cheung D, Li L, Dunia I, Benedetti LE, Horwitz J, Gong X (2006) Absence of alpha3 (Cx46) and alpha8 (Cx50) connexins leads to cataracts by affecting lens inner fiber cells. Exp Eye Res 83:688–696Google Scholar