Degenerative retinal diseases resulting from permanent loss of retinal neurons pose a great public health risk as no effective cell replacement therapies are currently available. De novo retinal regeneration from stem cells or retinal precursors could lead to a potential remedy for retinal degeneration diseases. Studies to identify genes and genetic processes essential for the formation of retinal neurons will provide the foundation for retinal regeneration. This proposal aims to reveal the molecular mechanisms governing the generation of retinal ganglion cells (RGCs), a group of neurons degenerated in glaucoma diseased eyes. The long-term objective of this proposal is to understand the role of an Atonal-class basic helix-loop-helix (bHLH) transcription factor, Math5, in the process of retinal neurogenesis and its interactions with other transcription factors in regulating the cell fate choices during retinal development. Knockout studies have shown that loss of Math5 leads to the absence of RGC formation and a concurrent increase in the generation of amacrine and cone photoreceptor cells, implying the dual roles of Math5 in promoting RGC differentiation pathway and inhibiting non-RGC differentiation pathways. Furthermore, we have shown that Math5 positively regulates the expression of RGC- specific transcription factors like Brn3b and Isl1, and negatively regulates non-RGC factors such as Bhlhb5, NeuroD, Math3, and Ngn2.
The Aim 1 of this proposal is designed to characterize the role of Bhlhb5 in the development of selective subtypes of amacrine and cone bipolar cells. Lineage analysis using Bhlhb5-Cre (Cre recombinase) mice will be carried out to test if Bhlhb5 expression defines amacrine and cone bipolar subtypes.
In Aim 2, the role of Isl1 in retinal genesis will be investigated in retina-specific Isl1 knockout mice. In addition, the synergistic effects of Math5 and Isl1 co-expression in promoting RGC formation from retinal precursors will be assessed by knocking-in Isl1 in Math5 locus.
The Aim 3 is proposed to test the regulatory relationship of Math5 and its known downstream target genes. Chromatin immunoprecipitation (ChIP) experiments using anti- HA and genetically modified Math5-HA mice will be used to identify the direct target genes of Math5 as well as to discover novel Math5 target genes. Alternatively, a Tamoxifen-inducible Math5 (Math5ER, a fusion of Math5 and estrogen receptor ligand-binding domain) will be used to identify genes directly activated by Math5 in the presence of Tamoxifen and protein synthesis inhibitor, cycloheximide. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
2R01EY013426-06A1
Application #
7371404
Study Section
Biology and Diseases of the Posterior Eye Study Section (BDPE)
Program Officer
Greenwell, Thomas
Project Start
2001-05-01
Project End
2010-11-30
Budget Start
2007-12-01
Budget End
2008-11-30
Support Year
6
Fiscal Year
2008
Total Cost
$366,100
Indirect Cost
Name
University of Rochester
Department
Ophthalmology
Type
Schools of Dentistry
DUNS #
041294109
City
Rochester
State
NY
Country
United States
Zip Code
14627
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Luo, Xiong-jian; Li, Ming; Huang, Liang et al. (2012) The interleukin 3 gene (IL3) contributes to human brain volume variation by regulating proliferation and survival of neural progenitors. PLoS One 7:e50375
Ding, Qian; Joshi, Pushkar S; Xie, Zheng-Hua et al. (2012) BARHL2 transcription factor regulates the ipsilateral/contralateral subtype divergence in postmitotic dI1 neurons of the developing spinal cord. Proc Natl Acad Sci U S A 109:1566-71
Xu, Hong-ping; Chen, Hui; Ding, Qian et al. (2010) The immune protein CD3zeta is required for normal development of neural circuits in the retina. Neuron 65:503-15
Feng, Liang; Xie, Zheng-hua; Ding, Qian et al. (2010) MATH5 controls the acquisition of multiple retinal cell fates. Mol Brain 3:36
Ding, Qian; Chen, Hui; Xie, Xiaoling et al. (2009) BARHL2 differentially regulates the development of retinal amacrine and ganglion neurons. J Neurosci 29:3992-4003
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
Elshatory, Yasser; Gan, Lin (2008) The LIM-homeobox gene Islet-1 is required for the development of restricted forebrain cholinergic neurons. J Neurosci 28:3291-7
Joshi, Pushkar S; Molyneaux, Bradley J; Feng, Liang et al. (2008) Bhlhb5 regulates the postmitotic acquisition of area identities in layers II-V of the developing neocortex. Neuron 60:258-72
Pan, Ling; Deng, Min; Xie, Xiaoling et al. (2008) ISL1 and BRN3B co-regulate the differentiation of murine retinal ganglion cells. Development 135:1981-90

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