The long-term goal of this application is to elucidate the role of transcription factor Pax6 in lens biology and disease. Pax6 is essential for lens formation and function. Mutations or deletions in PAX6 cause ocular diseases including aniridia, early onset cataract, Peters'anomaly, and autosomal dominant keratitis. Pax6 regulates the expression of numerous genes that range from lens structural proteins including crystallins to other transcription factors including MafA, c-Maf, and Brg1. Evidence exists that Pax6 proteins functionally interact with Maf and Brgl proteins. Expression of Pax6 target genes is controlled through either gene activation or gene repression. Crystallin gene expression is directed by interactions of Pax6 with Mafs together with co-activators including ASC-2. To understand the function of Pax6 in lens, this proposal will: (1) Determine the molecular mechanism by which Pax6 activates the alpha B- and represses gamma F-crystallin genes;(2) Elucidate the function of chromatin remodeling factor Brg1, and transcriptional co-activator ASC-2 in vivo in lens;and (3) Identify and characterize genes regulated by Pax6 that control lens induction, invagination of the lens placode, and formation of the lens vesicle.
These aims will be achieved using functional analysis of gene expression including chromatin immunoprecipitations, promoter mutagenesis, transfections, and protein-DNA assays. The genetic experiments will be performed in vivo using transgenic mice. The function of Brg1 in lens will be determined using lens-specific inactivation of Brgl combined with an overexpression of the dominant negative (DN) Brgl mutant. The DN mutant of ASC-2 will also be expressed in lens, as it is required for the expression of specific crystallins. Differential gene expression between normal and Pax6 homozygous mouse embryos will be assessed using cDNA microarrays, followed by temporal and spatial analysis of the expression of selected genes. Collectively, these studies will refine our understanding of the genetic network regulated by Pax6 and mechanisms whereby Pax6 controls this genetic network in lens.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY012200-10
Application #
7539902
Study Section
Special Emphasis Panel (ZRG1-AED (01))
Program Officer
Araj, Houmam H
Project Start
2000-01-01
Project End
2009-12-31
Budget Start
2009-01-01
Budget End
2009-12-31
Support Year
10
Fiscal Year
2009
Total Cost
$484,390
Indirect Cost
Name
Albert Einstein College of Medicine
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
110521739
City
Bronx
State
NY
Country
United States
Zip Code
10461
Esteban-Martínez, Lorena; Sierra-Filardi, Elena; McGreal, Rebecca S et al. (2017) Programmed mitophagy is essential for the glycolytic switch during cell differentiation. EMBO J 36:1688-1706
Cvekl, Ales; Zhang, Xin (2017) Signaling and Gene Regulatory Networks in Mammalian Lens Development. Trends Genet 33:677-702
Cvekl, Ales; Zhao, Yilin; McGreal, Rebecca et al. (2017) Evolutionary Origins of Pax6 Control of Crystallin Genes. Genome Biol Evol 9:2075-2092
Liu, Wei; Cvekl, Ales (2017) Six3 in a small population of progenitors at E8.5 is required for neuroretinal specification via regulating cell signaling and survival in mice. Dev Biol 428:164-175
Cavalheiro, Gabriel R; Matos-Rodrigues, Gabriel E; Zhao, Yilin et al. (2017) N-myc regulates growth and fiber cell differentiation in lens development. Dev Biol 429:105-117
Sun, Jian; Zhao, Yilin; McGreal, Rebecca et al. (2016) Pax6 associates with H3K4-specific histone methyltransferases Mll1, Mll2, and Set1a and regulates H3K4 methylation at promoters and enhancers. Epigenetics Chromatin 9:37
He, Shuying; Limi, Saima; McGreal, Rebecca S et al. (2016) Chromatin remodeling enzyme Snf2h regulates embryonic lens differentiation and denucleation. Development 143:1937-47
Lowe, Albert; Harris, Raven; Bhansali, Punita et al. (2016) Intercellular Adhesion-Dependent Cell Survival and ROCK-Regulated Actomyosin-Driven Forces Mediate Self-Formation of a Retinal Organoid. Stem Cell Reports 6:743-756
Menuchin-Lasowski, Yotam; Oren-Giladi, Pazit; Xie, Qing et al. (2016) Sip1 regulates the generation of the inner nuclear layer retinal cell lineages in mammals. Development 143:2829-41
Sun, Jian; Rockowitz, Shira; Chauss, Daniel et al. (2015) Chromatin features, RNA polymerase II and the comparative expression of lens genes encoding crystallins, transcription factors, and autophagy mediators. Mol Vis 21:955-73

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