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Identification and characterization of genes regulating zebrafish lens development

Award Number: R15EY013535
ORGANIZATION: NATIONAL EYE INSTITUTE
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 08/01/2001
PERIOD OF PERFORMANCE END DATE: 03/31/2027

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Identification and characterization of genes regulating zebrafish lens development - PROJECT SUMMARY/ABSTRACT The lens of the vertebrate eye becomes transparent by removing the nuclei and organelles of its inner fiber cells. Remarkably, these fiber cells must stay alive and functional without the ability to produce new proteins. Defects in fiber cell differentiation can contribute to lens cataract, the leading cause of blindness worldwide. While much has been learned about the molecular mechanisms that coordinate lens fiber cell development, there are likely many key players yet to be discovered. Our laboratory uses the zebrafish as an efficient screening system to identify novel genes playing a role in this process. We recently used CRISPR/Cas9 gene editing to show that damaging the transcription factor gene, cebpg, and RNA-binding protein genes, csde1 and c1qbp, leads to defects in lens development. Interestingly, in non-lens tissues these three genes play roles that would be relevant to lens development, such as regulating autophagy, protein stability and the stress response. We hypothesize that the protein products of these genes perform similar roles in the lens. In Aim 1 of this proposal, we will characterize the spatial expression of each gene across the developing lens of wild-type fish using RNA-scope, which detects specific mRNA molecules. We will also detail changes in nuclear and mitochondrial loss and the elongation of fiber cells. These experiments will allow us to identify the aspects of differentiation altered by the loss of each gene. Aim 2 of this proposal will use RNA-Seq and RT-qPCR to detail how the lens transcriptome is altered in each mutant. This approach will allow us to determine if the expression of known regulators of lens fiber cell differentiation mechanisms such as autophagy, denucleation or stress response, are changing, which would implicate them as part of the same signaling network as our mutated genes. The transcriptomic analysis will also identify new possible regulators of lens development, which we can then screen with our CRISPR/Cas9 system. Our laboratory is well placed to successfully conduct these proposed experiments, which will provide undergraduate students with excellent training in modern molecular and developmental biology techniques and bioinformatics analysis. Our findings will take advantage of the zebrafish lens system to address broad questions about how cells remodel during tissue differentiation.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2025 ( Subtotal = $0 )
2025	2024	ASHLAND UNIVERSITY	401 COLLEGE AVE	ASHLAND	OH	44804		USA	Vision Research	001	6	4/11/2025	COMPETING CONTINUATION	$46,430
2025	2020	ASHLAND UNIVERSITY	401 COLLEGE AVE	ASHLAND	OH	44804		USA	Vision Research	000	5	4/11/2025	COMPETING CONTINUATION	-$46,430
														Subtotal = $0

Issue Date FY: 2024 ( Subtotal = $306,457 )
2024	2024	ASHLAND UNIVERSITY	401 COLLEGE AVE	ASHLAND	OH	44804		USA	Vision Research	001	6	3/26/2024	COMPETING CONTINUATION	$306,457
2024	2020	ASHLAND UNIVERSITY	401 COLLEGE AVE	ASHLAND	OH	44804		USA	Vision Research	000	5	3/20/2024	COMPETING CONTINUATION	$0
														Subtotal = $306,457

Issue Date FY: 2023 ( Subtotal = $0 )
2023	2020	ASHLAND UNIVERSITY	401 COLLEGE AVE	ASHLAND	OH	44805	ASHLAND	USA	Vision Research	000	5	8/24/2023	COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2022 ( Subtotal = $0 )
2022	2016	ASHLAND UNIVERSITY	401 COLLEGE AVE	ASHLAND	OH	44805	ASHLAND	USA	Vision Research	000	4	6/27/2022	COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2020 ( Subtotal = $311,167 )
2020	2020	ASHLAND UNIVERSITY	401 COLLEGE AVE	ASHLAND	OH	44805	ASHLAND	USA	Vision Research	002	5	7/7/2020	COMPETING CONTINUATION	$311,167
2020	2016	ASHLAND UNIVERSITY	401 COLLEGE AVE	ASHLAND	OH	44805	ASHLAND	USA	Vision Research	000	4	4/29/2020	COMPETING CONTINUATION	$0
2020	2016	ASHLAND UNIVERSITY	401 COLLEGE AVE	ASHLAND	OH	44805	ASHLAND	USA	Vision Research	001	4	6/30/2020	COMPETING CONTINUATION	$0
														Subtotal = $311,167

Issue Date FY: 2016 ( Subtotal = $305,615 )
2016	2016	ASHLAND UNIVERSITY	401 COLLEGE AVENUE	ASHLAND	OH	44805	ASHLAND	USA	Vision Research	000	4	4/28/2016	COMPETING CONTINUATION	$305,615
														Subtotal = $305,615

Issue Date FY: 2011 ( Subtotal = $25,548 )
2011	2010	ASHLAND UNIVERSITY	401 COLLEGE AVENUE	ASHLAND	OH	44805	ASHLAND	USA	Biomedical Research and Research Training	000	3	8/15/2011	COMPETING CONTINUATION	$25,548
														Subtotal = $25,548

Issue Date FY: 2010 ( Subtotal = $158,996 )
2010	2010	ASHLAND UNIVERSITY	401 COLLEGE AVENUE	ASHLAND	OH	44805	ASHLAND	USA	Academic Research Enhancement Award	000	3	1/28/2010	COMPETING CONTINUATION	$158,996
														Subtotal = $158,996

Issue Date FY: 2005 ( Subtotal = $132,794 )
2005	2005	ASHLAND UNIVERSITY	401 COLLEGE AVENUE	ASHLAND	OH	44805	ASHLAND	USA	Academic Research Enhancement Award	000	2	9/19/2005	COMPETING CONTINUATION	$132,794
														Subtotal = $132,794

Issue Date FY: 2004 ( Subtotal = $4,400 )
2004	2004	ASHLAND UNIVERSITY	401 COLLEGE AVENUE	ASHLAND	OH	44805	ASHLAND	USA	Vision Research	001	1	9/17/2004	SUPPLEMENT FOR EXPANSION	$4,400
														Subtotal = $4,400

Issue Date FY: 2001 ( Subtotal = $91,800 )
2001	2001	ASHLAND UNIVERSITY	401 COLLEGE AVENUE	ASHLAND	OH	44805	ASHLAND	USA	Academic Research Enhancement Award	000	1	8/27/2001	NEW	$91,800
														Subtotal = $91,800

Grand Total All Awards = $1,336,777

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Identification and characterization of genes regulating zebrafish lens development

Award Number: R15EY013535

ORGANIZATION: NATIONAL EYE INSTITUTE

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 08/01/2001

PERIOD OF PERFORMANCE END DATE: 03/31/2027

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