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1 From the Department of Physiology and Biophysics, State University of New York, Stony Brook 2 Departments of Neurobiology and 3 Cell Biology, Harvard Medical School, Boston, Massachusetts.
PURPOSE. To determine the roles of intercellular communication in embryonic eye growth and development, mice with a targeted deletion of the Cx43 gene were examined, and mice without both Cx43 and Cx50 were generated and analyzed.
METHODS. Embryonic eyes and lenses from wild-type mice, or mice deficient in Cx43, Cx50, or both Cx43 and Cx50 were collected and analyzed structurally by light and electron microscopy, immunohistochemically using connexin-specific antibodies, biochemically by Western blot analysis, and physiologically by measuring patterns of junctional communication revealed by iontophoretic injection of junction-permeable reporter molecules.
RESULTS. Cx50 expression was limited to the ocular lens and was
not detected in either the cornea or the retina.
Cx43-/- embryos showed development of
structurally normal lenses and eyes when examined by light and electron
microscopy through embryonic day (E)18.5. In addition,
Cx43-/- lenses synthesized four different
markers of lens differentiation: MIP26, 
A-crystallin,
B-crystallin, and 
-crystallin. Double-knockout lenses were also
histologically normal through E18.5 and synthesized the four lens
differentiation markers. When assayed by intracellular injection with
Lucifer yellow (Molecular Probes, Eugene, OR) and neurobiotin
at E15.5,
Cx43-/-/Cx50-/-
lenses retained gap junction–mediated dye transfer between fiber
cells. In contrast, dye transfer in double-knockout lenses was
dramatically reduced between epithelial cells and was eliminated
between epithelial cells and fibers.
CONCLUSIONS. These data indicate that the unique functional properties of both Cx43 and Cx50 are not required for prenatal lens development and that connexin diversity is required for regulation of postnatal growth and homeostasis.
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