Ocular lens morphogenesis is a model for investigating mechanisms of cellular differentiation, spatial and temporal gene expression control, and chromatin regulation. Brg1 (Smarca4) and Snf2h (Smarca5) are catalytic subunits of distinct ATP-dependent chromatin remodeling complexes implicated in transcriptional regulation. Previous studies have shown that Brg1 regulates both lens fiber cell differentiation and organized degradation of their nuclei (denucleation). Here, we employed a conditional Snf2h(flox) mouse model to probe the cellular and molecular mechanisms of lens formation. Depletion of Snf2h induces premature and expanded differentiation of lens precursor cells forming the lens vesicle, implicating Snf2h as a key regulator of lens vesicle polarity through spatial control of Prox1, Jag1, p27(Kip1) (Cdkn1b) and p57(Kip2) (Cdkn1c) gene expression. The abnormal Snf2h(-/-) fiber cells also retain their nuclei. RNA profiling of Snf2h(-/) (-) and Brg1(-/-) eyes revealed differences in multiple transcripts, including prominent downregulation of those encoding Hsf4 and DNase IIβ, which are implicated in the denucleation process. In summary, our data suggest that Snf2h is essential for the establishment of lens vesicle polarity, partitioning of prospective lens epithelial and fiber cell compartments, lens fiber cell differentiation, and lens fiber cell nuclear degradation.

1st Faculty of Medicine Charles University 121 08 Prague Czech Republic

Department of Biomedical Science Florida Atlantic University Boca Raton FL 33431 USA

Department of Cell Biology Albert Einstein College of Medicine Bronx NY 10461 USA

Department of Cell Biology Albert Einstein College of Medicine Bronx NY 10461 USA 1st Faculty of Medicine Charles University 121 08 Prague Czech Republic

Department of Human Molecular Genetics and Biochemistry Sackler School of Medicine Tel Aviv University Ramat Aviv Tel Aviv 69978 Israel

Department of Ophthalmology and Visual Sciences and Genetics Albert Einstein College of Medicine Bronx NY 10461 USA

Department of Pathology and NYU Center for Health Informatics and Bioinformatics New York University Langone Medical Center New York NY 10016 USA Mechanisms of Carcinogenesis Section International Agency for Research on Cancer Lyon Cedex 08 69372 France

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