Further analysis of the lens phenotype in Lim2-deficient mice.

Yanrong Shi, Alicia B De Maria, Huan Wang, Richard T Mathias, Paul G FitzGerald, Steven Bassnett

Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, St. Louis, Missouri 63110, USA.

Investigative ophthalmology & visual science 2011 Sep

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Lim2 (MP20) is the second most abundant integral protein of lens fiber cell membranes. A comparative analysis was performed of wild-type and Lim2-deficient (Lim2(Gt/Gt)) mouse lenses, to better define the anatomic and physiologic roles of Lim2. Scanning electron microscopy (SEM) and confocal microscopy were used to assess the contribution of Lim2 to lens tissue architecture. Differentiation-dependent changes in cytoskeletal composition were identified by mass spectrometry and immunoblot analysis. The effects on cell-cell communication were quantified using impedance analysis. Lim2-null lenses were grossly normal. At the cellular level, however, subtle structural alterations were evident. Confocal microscopy and SEM analysis revealed that cortical Lim2(Gt/Gt) fiber cells lacked the undulating morphology that characterized wild-type fiber cells. On SDS-PAGE analysis the composition of cortical fiber cells from wild-type and Lim2-null lenses appeared similar. However, marked disparities were evident in samples prepared from the lens core of the two genotypes. Several cytoskeletal proteins that were abundant in wild-type core fiber cells were diminished in the cores of Lim2(Gt/Gt) lenses. Electrophysiological measurements indicated a small decrease in the membrane potential of Lim2(Gt/Gt) lenses and a two-fold increase in the effective intracellular resistivity. In the lens core, this may have reflected decreased expression levels of the gap junction protein connexin 46 (Cx46). In contrast, increased resistivity in the outer cell layers of Lim2(Gt/Gt) lenses could not be attributed to decreased connexin expression and may reflect the absence of cell fusions in Lim2(Gt/Gt) lenses. Comparative analysis of wild-type and Lim2-deficient lenses has implicated Lim2 in maintenance of cytoskeletal integrity, cell morphology, and intercellular communication.

Citation

Yanrong Shi, Alicia B De Maria, Huan Wang, Richard T Mathias, Paul G FitzGerald, Steven Bassnett. Further analysis of the lens phenotype in Lim2-deficient mice. Investigative ophthalmology & visual science. 2011 Sep;52(10):7332-9