The surface ectoderm directly overlying the developing optic vesicle gives rise to two stratified epithelial tissues of the eye, the corneal epithelium and the crystalline lens. Early morphogenesis of the lens vesicle and further development and differentiation of the ocular epithelia involve tightly regulated mechanics of cell determination and differentiation. the nature and specific role of regulatory molecules controlling these gene expression programs are largely unknown. Recent evidence in the literature suggests that a family of transcription factors known as AP-2 (Activating Protein - 2) factors, are necessary for craniofacial morphogenesis. preliminary findings from this laboratory suggest that transcription factor AP-2 (Activating Protein 2) plays an important role in eye development, specifically regulating genes involved in morphogenesis of the lens and corneal epithelium. The proposed study will test the hypothesis that AP-2 transcription factors regulate gene expression programs that determine: a) early pattern formation of the lens vesicle and b) later stages of development and differentiation of the corneal epithelium and ocular lens. Determination of the expression pattern of three AP-2 proteins (a, B, y) in the differentiating and regenerating ocular epithelia using immunolocalization and in situ hybridization techniques will enable us to compare with the location of candidate downstream genes. To investigate the specific functional requirement of the AP-2a gene during morphogenesis of the lens vesicle, a detailed morphological examination of the ocular defects of AP-2a-/- knockout mice will be performed. In addition, the newly derived Lens complementation system (LCS) will be used to directly determine whether AP-2A is required for formation of the lens. To determine the specific role of AP-2 factors during lens fiber cell and corneal epithelial differentiation. AP-2 activity will be targeted in transgenic mice and in vitro culture models. Using a wild-type P-2 minigene and dominant negative minigenes designed to interfere with AP-2 activity, AP-2 activity will be altered in living cells in vitro and in transgenic mice in order to determine how AP-2 regulates specific aspects of epithelial cell biology, including cell adhesion.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY011910-02
Application #
2882936
Study Section
Visual Sciences A Study Section (VISA)
Project Start
1998-03-01
Project End
2002-02-28
Budget Start
1999-03-01
Budget End
2000-02-29
Support Year
2
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Tufts University
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02111
Bassett, Erin A; Korol, Anna; Deschamps, Paula A et al. (2012) Overlapping expression patterns and redundant roles for AP-2 transcription factors in the developing mammalian retina. Dev Dyn 241:814-29
Kerr, Christine L; Huang, Jian; Williams, Trevor et al. (2012) Activation of the hedgehog signaling pathway in the developing lens stimulates ectopic FoxE3 expression and disruption in fiber cell differentiation. Invest Ophthalmol Vis Sci 53:3316-30
Bassett, Erin A; Williams, Trevor; Zacharias, Amanda L et al. (2010) AP-2alpha knockout mice exhibit optic cup patterning defects and failure of optic stalk morphogenesis. Hum Mol Genet 19:1791-804
Pontoriero, Giuseppe F; Smith, April N; Miller, Leigh-Anne D et al. (2009) Co-operative roles for E-cadherin and N-cadherin during lens vesicle separation and lens epithelial cell survival. Dev Biol 326:403-17
Pontoriero, Giuseppe F; Deschamps, Paula; Ashery-Padan, Ruth et al. (2008) Cell autonomous roles for AP-2alpha in lens vesicle separation and maintenance of the lens epithelial cell phenotype. Dev Dyn 237:602-17
Bassett, Erin A; Pontoriero, Giuseppe F; Feng, Weiguo et al. (2007) Conditional deletion of activating protein 2alpha (AP-2alpha) in the developing retina demonstrates non-cell-autonomous roles for AP-2alpha in optic cup development. Mol Cell Biol 27:7497-510
Makhani, Leila F; Williams, Trevor; West-Mays, Judith A (2007) Genetic analysis indicates that transcription factors AP-2alpha and Pax6 cooperate in the normal patterning and morphogenesis of the lens. Mol Vis 13:1215-25
West-Mays, Judith A; Dwivedi, Dhruva J (2006) The keratocyte: corneal stromal cell with variable repair phenotypes. Int J Biochem Cell Biol 38:1625-31
Dwivedi, Dhruva J; Pontoriero, Giuseppe F; Ashery-Padan, Ruth et al. (2005) Targeted deletion of AP-2alpha leads to disruption in corneal epithelial cell integrity and defects in the corneal stroma. Invest Ophthalmol Vis Sci 46:3623-30
Sivak, Jeremy M; West-Mays, Judith A; Yee, Amy et al. (2004) Transcription Factors Pax6 and AP-2alpha Interact To Coordinate Corneal Epithelial Repair by Controlling Expression of Matrix Metalloproteinase Gelatinase B. Mol Cell Biol 24:245-57

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