In the mouse visual system, axons of retinal ganglion cells (RGCs) exit from each eye and converge at the ventral midline of the brain to form the optic chiasm (OC), then grow to visual centers in the thalamus, where they synapse onto second-order neurons that project to the cerebral cortex. In animals with frontally located eyes, some RGC axons do not cross the midline and instead project to the ipsilateral, or same-side, visual centers. This circuit mediates binocular vision and we have long focused on the formation of this circuit. The present proposal addresses how ipsilateral (ipsi) and contralateral (contra) RGCs axons extend together in common tracts through the developing brain, converging and diverging throughout their journey to the target, and whether such cohort integrity in paths is relevant to connectivity within target regions. In this proposal, we seek to more fully characterize eye-specific axonal cohort maneuvers in the pathway from eye through the OC to first target.
In Aim 1, we will investigate the organization of the ipsi and contra axons, viewed with different tracers in the same preparation, focusing on where the ipsi and contra RGC axons from the same and opposite eyes interleave and where they segregate. We will use classical and novel morphological analysis at the LM and EM levels to chart fasciculation of ipsi and contra RGCs along the path and their relationships with glial cells. We will also devise explant assays to dissect the cellular and molecular mechanisms for fiber-fiber interactions during eye-specific axon pathway formation.
In Aim 2, we will investigate the effects of mutations that have known effects on decussation and targeting on RGC eye- specific fiber organization. Throughout, we will compare and contrast the cell and molecular differences of ipsi and contra RGCs as they diverge amongst radial glia at the optic chiasm midline, associate with their own and opposite eye cohorts, exit the chiasm and grow in the tract where astroglia are organized to potentially influence their trajectory, and enter and terminate in the dorsal lateral geniculate nuclei (dLGN).. Approaches developed over the last few decades in our lab will facilitate analysis of axons at the single fiber and cohort levels, aided by a newly developed clearing technique and special microscopy, and electroporation in utero for delivery of GFP-constructs into the retina. We will address the relationship between decussation, eye-specific organization within the path, and early target innervation, linking phases of axon guidance that were previously addressed as discrete steps and restricted to focus on a single cohort. Understanding how tracts are laid down is essential for unraveling the phenotypes of neurological disorders, in which defects in fiber pathways and synapse formation are implicated but their relationship is not fully understood. Moreover, implementing axon regeneration depends on knowledge of how the entire pathway is organized and to what extent each phase of axon organization prepares axons for the next phase.

Public Health Relevance

Proper binocular vision depends on a normal distribution of retinal axons at the optic chiasm, and if altered, reduced visual acuity and depth perception ensue. This research aims to understand how growing retinal ganglion cells (RGCs) from each eye converge at the X-shaped optic chiasm, and then diverge toward targets on the same and opposite side of the brain, coursing in segregated eye-specific bundles before entering and innervating targets in distinct synaptic territories. Information on the mechanisms underlying this changing organization, both intrinsic to RGCs and extrinsic from glia along the path, and the relationship of fiber organization to eye-specific targeting is key to understanding neurodevelopmental disorders, many of which display fiber tract alterations.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY012736-27
Application #
9136842
Study Section
Neurodifferentiation, Plasticity, and Regeneration Study Section (NDPR)
Program Officer
Greenwell, Thomas
Project Start
1999-07-01
Project End
2018-07-31
Budget Start
2016-08-01
Budget End
2017-07-31
Support Year
27
Fiscal Year
2016
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Pathology
Type
Schools of Medicine
DUNS #
621889815
City
New York
State
NY
Country
United States
Zip Code
10032
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Sitko, Austen A; Kuwajima, Takaaki; Mason, Carol A (2018) Eye-specific segregation and differential fasciculation of developing retinal ganglion cell axons in the mouse visual pathway. J Comp Neurol 526:1077-1096
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Soares, Célia A; Mason, Carol A (2015) Transient ipsilateral retinal ganglion cell projections to the brain: Extent, targeting, and disappearance. Dev Neurobiol 75:1385-401

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