The long term goal of this research is to elucidate molecular mechanisms by which cell-extrinsic cues influence retinal progenitor cell fate choices and differentiation during normal development, thereby providing a basis for the prevention and treatment of retinal diseases. The proposed study focuses on the cytokine-signaling pathway in the mouse retina. Ciliary neurotrophic factor (CNTF), a member of the cytokine superfamily, has a variety of effects on neuronal development and survival. In the developing retina, CNTF profoundly affects photoreceptor, bipolar, and amacrine cell differentiation as well as promoting ganglion cell axonal growth and survival. In addition, CNTF prevents retinal photoreceptor cell degeneration caused by mutations or light-induced damages. Despite these effects, the mechanism of CNTF action in the developing or mature retina is not understood. Cytokine signals are generally mediated through membrane receptors that activate cellular Jak tyrosine kinases and STAT transcription factors. However, the specific signaling components in the retina that mediate the effects of CNTF, and the retinal cell types that directly respond to the CNTF signal, have not been characterized. Therefore, experiments are proposed to biochemically identify the Jak kinases and STAT transcription factors activated by CNTF in the developing mouse retina using immunoprecipitation and Western blot analyses of retinal cell extracts. The distribution and cellular location of the CNTF receptor, Jak protein kinases and STAT factors in the retina will be determined by immunocytochemistry to define retinal cell types that can respond to CNTF or CNTF-like cytokines. In addition, cell type-specific activation of STAT transcription factors will be assayed by immunofluorescence microscopy of dissociated cell and explant cultures treated with CNTF using combinations of antibodies recognizing activated (phosphorylated) STAT factors and distinct retinal cell types. Lastly, the biological function of the cytokine signaling pathway during retinal neurogenesis will be examined by perturbing the activity of Jak and STAT signaling molecules. Embryonic and postnatal retinas will be infected in vivo and in vitro with murine retroviral vectors expressing mutant Jak kinases and STAT factors, and naturally-occurring Jak and STAT inhibitor proteins. Effects of these perturbations on retinal development will be examined with histological and immunocytochemical methods using molecular and cellular markers. These proposed studies will elucidate the biological function and cellular mechanisms of cytokine signal transduction during mammalian retinal neurogenesis and thus establish a foundation for therapeutic interventions of human retinal diseases using CNTF or CNTF-like cytokines.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
1R01EY012270-01A2
Application #
6132607
Study Section
Visual Sciences C Study Section (VISC)
Program Officer
Hunter, Chyren
Project Start
2000-05-01
Project End
2004-04-30
Budget Start
2000-05-01
Budget End
2001-04-30
Support Year
1
Fiscal Year
2000
Total Cost
$264,464
Indirect Cost
Name
University of California Los Angeles
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Rhee, K-D; Yu, J; Zhao, C Y et al. (2012) Dnmt1-dependent DNA methylation is essential for photoreceptor terminal differentiation and retinal neuron survival. Cell Death Dis 3:e427
Sakagami, Kiyo; Chen, Bryan; Nusinowitz, Steven et al. (2012) PTEN regulates retinal interneuron morphogenesis and synaptic layer formation. Mol Cell Neurosci 49:171-83
Rhee, Kun Do; Yang, Xian-Jie (2010) Function and mechanism of CNTF/LIF signaling in retinogenesis. Adv Exp Med Biol 664:647-54
Sakagami, Kiyo; Gan, Lin; Yang, Xian-Jie (2009) Distinct effects of Hedgehog signaling on neuronal fate specification and cell cycle progression in the embryonic mouse retina. J Neurosci 29:6932-44
Yang, Xin; Zhang, Hua; Zhang, Yingchun et al. (2008) Two new diterpenoid acids from Pinus koraiensis. Fitoterapia 79:179-81
Goureau, Olivier; Rhee, Kun Do; Yang, Xian-Jie (2004) Ciliary neurotrophic factor promotes muller glia differentiation from the postnatal retinal progenitor pool. Dev Neurosci 26:359-70
Yang, Xian-Jie (2004) Roles of cell-extrinsic growth factors in vertebrate eye pattern formation and retinogenesis. Semin Cell Dev Biol 15:91-103
Rhee, Kun Do; Goureau, Olivier; Chen, Shiming et al. (2004) Cytokine-induced activation of signal transducer and activator of transcription in photoreceptor precursors regulates rod differentiation in the developing mouse retina. J Neurosci 24:9779-88
Zhang, Xian-Mei; Yang, Zhenglin; Karan, Goutam et al. (2003) Elovl4 mRNA distribution in the developing mouse retina and phylogenetic conservation of Elovl4 genes. Mol Vis 9:301-7
Hashimoto, Takao; Zhang, Xiang-Mei; Yang, Xian-Jie (2003) Expression of the Flk1 receptor and its ligand VEGF in the developing chick central nervous system. Gene Expr Patterns 3:109-13

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