The long term goal of this proposal is to investigate the genetic mechanisms and microenvironmental factors regulating the survival and differentiated function of retinal neurons and photoreceptors. The strategy used is based on comparative analysis of the differentiated fate of chick embryo retinal precursor cells as they develop either within their normal microenvironment in vivo, or under experimentally-controlled conditions in dissociated cultures. Terminal mitosis of the cells is determined by combined thymidine autoradiography and bromodeoxyuridine immunocytochemistry, and cell differentiation in vivo and in vitro is characterized using a multidisciplinary approach including immunocytochemistry, in situ hybridization, Northern blot analysis and the polymerase chain reaction. The mechanisms that control the differentiation of retinal precursor cells as either rod or cone photoreceptors will be studied by investigating the expression of the genes for visual pigments and other photoreceptor-specific molecules in vivo. Other experiments are proposed to analyze the expression of the homeobox genes Pax6, Prox1, Chx10, and Brn3 in developing retinal cells in vivo and in vitro, and their overexpression in cultured retinal cells using retroviral vectors. These studies are expected to shed light on cellular and molecular mechanisms of normal retinal development and function, and to contribute to the search for therapeutic approaches for retinal degenerative disorders.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY004859-17
Application #
2888144
Study Section
Special Emphasis Panel (ZRG1-VISC (01))
Project Start
1983-01-01
Project End
2001-06-30
Budget Start
1999-07-01
Budget End
2000-06-30
Support Year
17
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Johns Hopkins University
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Vergara, M Natalia; Gutierrez, Christian; Canto-Soler, M Valeria (2015) Efficient Gene Transfer in Chick Retinas for Primary Cell Culture Studies: An Ex-ovo Electroporation Approach. J Vis Exp :e52002
Springelkamp, Henriƫt; Mishra, Aniket; Hysi, Pirro G et al. (2015) Meta-analysis of Genome-Wide Association Studies Identifies Novel Loci Associated With Optic Disc Morphology. Genet Epidemiol 39:207-16
Vergara, M Natalia; Gutierrez, Christian; O'Brien, David R et al. (2013) Ex vivo electroporation of retinal cells: a novel, high efficiency method for functional studies in primary retinal cultures. Exp Eye Res 109:40-50
Vergara, M Natalia; Canto-Soler, M Valeria (2012) Rediscovering the chick embryo as a model to study retinal development. Neural Dev 7:22
Vergara, M Natalia; de la Rosa, Enrique J; Canto-Soler, M Valeria (2012) Focus on molecules: proinsulin in the eye: precursor or pioneer? Exp Eye Res 101:109-10
Gutierrez, Christian; McNally, Minda; Canto-Soler, M Valeria (2011) Cytoskeleton proteins previously considered exclusive to ganglion cells are transiently expressed by all retinal neuronal precursors. BMC Dev Biol 11:46
McNally, Minda M; Wahlin, Karl J; Canto-Soler, M Valeria (2010) Endogenous expression of ASLV viral proteins in specific pathogen free chicken embryos: relevance for the developmental biology research field. BMC Dev Biol 10:106
Wahlin, Karl J; Hackler Jr, Laszlo; Adler, Ruben et al. (2010) Alternative splicing of neuroligin and its protein distribution in the outer plexiform layer of the chicken retina. J Comp Neurol 518:4938-62
Huang, Hu; Wahlin, Karl J; McNally, Minda et al. (2008) Developmental regulation of muscleblind-like (MBNL) gene expression in the chicken embryo retina. Dev Dyn 237:286-96
Wahlin, Karl J; Moreira, Ernesto F; Huang, Hu et al. (2008) Molecular dynamics of photoreceptor synapse formation in the developing chick retina. J Comp Neurol 506:822-37

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