Rajendra K Gangalum1, Zhe Jing1, Ankur M Bhat1, Josh Lee2, Yoshiko Nagaoka3, Sophie X Deng4, Meisheng Jiang3, Suraj P Bhat5. 1. Jules Stein Eye Institute, Geffen School of Medicine at University of California Los Angeles, Los Angeles, California, United States. 2. Jules Stein Eye Institute, Geffen School of Medicine at University of California Los Angeles, Los Angeles, California, United States Department of Molecular and Medical Pharmacology, Geffen School of Medicine at University of California Los Angeles, Los Angeles, California, United States Molecular Biology Institute and Brain Research Institute, University of California Los Angeles, Los Angeles, California, United States. 3. Department of Molecular and Medical Pharmacology, Geffen School of Medicine at University of California Los Angeles, Los Angeles, California, United States. 4. Molecular Biology Institute and Brain Research Institute, University of California Los Angeles, Los Angeles, California, United States. 5. Jules Stein Eye Institute, Geffen School of Medicine at University of California Los Angeles, Los Angeles, California, United States Molecular Biology Institute and Brain Research Institute, University of California Los Angeles, Los Angeles, California, United States.
Abstract
PURPOSE: The clinical management of cataracts in infancy involves surgical removal of the lens to ensure transmission of light to the retina, which is essential for normal neural development of the infant. This surgery, however, entails a lifelong follow-up and impaired vision. To our knowledge, no animal models recapitulate human lamellar opacities, the most prevalent form of early childhood cataracts. We present data on the recreation of the human lamellar cataract phenotype in transgenic mice. METHODS: Mutations in the DNA binding domain (DBD) of the heat shock transcription factor 4 (HSF4) are known to be associated with early childhood autosomal dominant lamellar cataract. We used bacterial artificial chromosome (BAC) transgenesis to express a hybrid gene: Hsf4 (DBD)-enhanced green fluorescent protein (EGFP), by recombineering EGFP sequences into the DBD of the Hsf4 gene, to interfere with the DNA binding properties of Hsf4. RESULTS: We recapitulated the human lamellar cataract, in its temporal as well as spatial presentation, within the transgenic mouse lens. This phenotype was reproduced faithfully using four different BACs, indicating that EGFP can be used to target transcription factor function in transgenic mice. Molecular and cell biological examination of early postnatal transgenic lens reveals impairment of secondary fiber cell differentiation. CONCLUSIONS: Recreation of the human lamellar cataract phenotype in mice allows investigation of this human pathology at a level not possible previously and points to the relevance of fiber cell heterogeneity dictated by fiber cell-specific gene activity in the biogenesis of the lamellar cataract. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.
PURPOSE: The clinical management of cataracts in infancy involves surgical removal of the lens to ensure transmission of light to the retina, which is essential for normal neural development of the infant. This surgery, however, entails a lifelong follow-up and impaired vision. To our knowledge, no animal models recapitulate humanlamellar opacities, the most prevalent form of early childhood cataracts. We present data on the recreation of the humanlamellar cataract phenotype in transgenic mice. METHODS: Mutations in the DNA binding domain (DBD) of the heat shock transcription factor 4 (HSF4) are known to be associated with early childhood autosomal dominant lamellar cataract. We used bacterial artificial chromosome (BAC) transgenesis to express a hybrid gene: Hsf4 (DBD)-enhanced green fluorescent protein (EGFP), by recombineering EGFP sequences into the DBD of the Hsf4 gene, to interfere with the DNA binding properties of Hsf4. RESULTS: We recapitulated the humanlamellar cataract, in its temporal as well as spatial presentation, within the transgenicmouse lens. This phenotype was reproduced faithfully using four different BACs, indicating that EGFP can be used to target transcription factor function in transgenic mice. Molecular and cell biological examination of early postnatal transgenic lens reveals impairment of secondary fiber cell differentiation. CONCLUSIONS: Recreation of the humanlamellar cataract phenotype in mice allows investigation of this human pathology at a level not possible previously and points to the relevance of fiber cell heterogeneity dictated by fiber cell-specific gene activity in the biogenesis of the lamellar cataract. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.
Authors: Helmut Hofemeister; Giovanni Ciotta; Jun Fu; Philipp Martin Seibert; Alexander Schulz; Marcello Maresca; Mihail Sarov; Konstantinos Anastassiadis; A Francis Stewart Journal: Methods Date: 2010-12-31 Impact factor: 3.608
Authors: Narsing A Rao; Sindhu Saraswathy; Guey Shuang Wu; George S Katselis; Eric F Wawrousek; Suraj Bhat Journal: Invest Ophthalmol Vis Sci Date: 2008-03 Impact factor: 4.799