Cell Biology, Neurobiology & Anatomy

Brian A. Link, PhD
Associate Professor

Department of Cell Biology, Neurobiology & Anatomy
Medical College of Wisconsin
8701 Watertown Plank Road
Milwaukee, WI 53226-0509

Phone: (414) 456-8072
FAX: (414) 456-6517
email: blink@mcw.edu

Mechanisms of vertebrate retinogenesis

There are several interrelated and ongoing projects for this topic. Emphasis has been placed on identifying regulators of retinogenesis by analyzing zebrafish mutants that show disrupted retinal lamination (Figure 1).

This phenotype is analogous to the "rough-eye" phenotype of the Drosophila omatidium which has been so critical in understanding the principles of signaling during development. Characterizations of zebrafish mutants have shed light on fundamental processes of retinogenesis including regulation and selection of cell cycle exit, post-mitotic cell migration, and the mechanisms of coordinated cellular differentiation. Additional emphasis in our lab is now being placed on studying the process of interkinetic nuclear migration and its relationships to neurogenesis. Interkinetic nuclear migration is the cellular behavior where neuroepithelia nuclei migrate in an apical-basal manner in phase with the cell cycle. The process is heterogeneous among the progenitors. Confocal time-lapse imaging with cell type specific GFP-based transgenes revealed that particular patterns of interkinetic nuclear migration influence which progenitor cells exit the cell cycle (Baye and Link, manuscript in revision) (Figure 2 and Movie 1).

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Movie 1. In vivo confocal time-lapse movie shows heterogeneity of interkinetic nuclear migration in retinal progenitors from 24 hpf. Images are composed of compressed z-stacks of H2A-GFP fluorescence overlaid with single mid-plane bright-field images. Apical and basal surfaces are outlined with dashed white lines in the first and last frame and an arrow indicates a cell undergoing mitosis at the apical surface. The movie plays at 6 frames per second covering 12 hr of development time (Baye and Link, 2007).

Specifically, neuroepithelia with deep basal nuclear migrations and shorter cell cycles are biased to produce neurons, rather than give rise to daughter cells that re-enter the cell cycle. The lab is currently exploring the signals and mechanisms underlying these relationships.

Complex gene interactions underlying glaucoma-phenotypes

The overall goal of these studies is to understand the relationships between genes that contribute to glaucoma-phenotypes. The glaucomas are a heterogeneous group of ocular disorders characterized by retinal ganglion cell death, optic nerve damage and visual field loss. Elevated intraocular pressure is a principal risk factor for this neurodegenerative disease. The majority and perhaps all forms of glaucoma are complex – requiring the interaction of multiple genes. While several genes that contribute to glaucoma have been identified, many additional loci remain unknown. We are using zebrafish to identify and study genes which promote glaucoma-phenotypes (Figure 3).

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Skarie, JM and Link, BA: The primary open-angle glaucoma gene WDR36 functions in ribosomal-RNA processing and interacts with the p53 stress-response pathway. Human Molecular Genetics. In press.

Berry, FB, *Skarie, JM, Mirzayans, F, Hudson, TJ, Raymond, V, Lind, BA, Walter, MA: FOXC1 is required for cell viability and resistance to oxidative stress in the eye through the transcriptional regulation of FOXO1A. Human Molecular Genetics 17:490-505, 2008.

Baye, LM and Link, BA: Nuclear migration during retinal development. Brain Research 1192:29-36, 2008.

Baye, LM and Link, BA: Interkinetic nuclear migration and the selection of neurogenic cell divisions during vertebrate retinogenesis. J Neuroscience 27:10143-52, 2007.

Baye, LM and Link, BA: The disarrayed mutation results in cell cycle and neurogenesis defects during retinal development in zebrafish. BMC Developmental Biology 7:28, 2007.

Cui S, C Otten, S Rohr, S Abdelilah-Seyfried and BA Link: Analysis of aPKClamda and aPKCzeta reveals multiple and redundant functions during vertebrate retinogenesis. Molecular Cellular Neuroscience 34(3):431-444, 2007.

Tamimi Y, JM Skarie, T Footz, FB Berry, BA Link and MA Walter: FGF19 is a target for FOXC1 regulation in ciliary body derived cells. Human Molecular Genetics 15:3229-40, 2006.

Semina EV, DV Bosenko, NA Zinkevich, KA Soules, DR Hyde, TC Vihtelic, GB Willer, RG Gregg and BA Link: Mutations in laminin alpha 1 result in complex ocular phenotypes in zebrafish. Developmental Biology 299:63-77, 2006.

Willer GB, VM Lee, RG Gregg and BA Link: Analysis of the zebrafish perplexed mutation reveals tissue specific roles for de novo pyrimidine synthesis during development. Genetics 170:1827-1837, 2005.

Soules KA and BA Link: Morphogenesis of the anterior segment in the zebrafish eye. BMC Developmental Biology 5:12, 2005.

Kay JN, BA Link and H Baier: Staggered, cell-intrinsic timing of ath5 expression underlies the wave of ganglion cell neurogenesis in the zebrafish retina. Development 132:2573-2585, 2005.

Link BA, M Gray, R Smith and S John: Intraocular pressure in zebrafish: comparison of inbred strains and identification of a reduced melanin mutant with raised IOP. Invest Ophthal Vision Sci 45:4415-4422, 2004.

McMahon C, E Semina and B Link: Using zebrafish to study the complex genetics of glaucoma. Comparative Biochemistry and Physiology 138:343-350, 2004.

Gregg RG, GB Willer, JM Fadool, JD Dowling and BA Link: Positional cloning of the young mutation identifies an essential role for the Brahma chromatin remodeling complex in mediating retinal cell differentiation. PNAS 100:6535-6540, 2003.