Myelin is a fundamental component of mature neural networks that is affected in a large number of pathological conditions of the central nervous system (CNS). Critical for advancing knowledge about these conditions would be a better in vivo understanding of how oligodendrocytes and their respective myelin sheaths develop, are maintained throughout life and respond to injury. Growing evidence suggests neuronal activity and proper astrocyte function may play crucial roles in these processes. We have developed a new technique that allows high resolution label-free in vivo imaging of myelinated axons. This technique takes advantage of the high refractive index of lipid rich multilayered myelin and is based on spectral confocal reflectance (SCoRe) microscopy. Using SCoRe imaging I have obtained for the first time long-term images of the dynamics of cortical myelin on the cellular scale in a living animal. Preliminary data shows this technique as well as fluorescence imaging of oligodendrocytes, axons and astrocytes are a powerful set of tools that in combination provide a wealth of information about fine structural dynamics of these structures in vivo. I demonstrate the feasibility to track long- term changes in internode length in addition to documenting the temporal dynamics of demyelination after single oligodendrocyte and astrocyte ablation. I propose to use these powerful techniques to address three fundamental questions concerning the in vivo plasticity and regeneration of myelin and oligodendrocytes. First, I will determine the long term plasticity of myelin and oligodendrocytes in the mouse cortex. Next I will determine the effects of neuronal activity on myelin formation, plasticity and oligodendrocyte regeneration. Finally I will use single cell ablation techniques to determine if astrocytes are required for or alter the temporal dynamics of oligodendrocyte remyelination in vivo. Together these experiments will describe for the first time the longitudinal dynamics and regeneration capabilities of oligodendrocytes and myelin on the cellular scale. Furthermore these experiments will reveal how changes in axons and astrocytes influence these dynamics over weeks to months in the living brain.
Myelination is a critical functional component of the central nervous system circuitry. Our limited knowledge of the in vivo dynamics of myelin and oligodendrocytes impedes our ability to understand how these structures are affected in debilitating demyelinating and neurodegenerative diseases. In this fellowship I propose to use a novel in vivo myelin imaging technique that we recently developed in combination with other cellular in vivo manipulation and imaging methods to determine the long-term plasticity, neuronal activity dependence, and dynamics of regeneration of neocortical myelin and oligodendrocytes.
|Hill, Robert A; Damisah, Eyiyemisi C; Chen, Fuyi et al. (2017) Targeted two-photon chemical apoptotic ablation of defined cell types in vivo. Nat Commun 8:15837|
|Damisah, Eyiyemisi C; Hill, Robert A; Tong, Lei et al. (2017) A fluoro-Nissl dye identifies pericytes as distinct vascular mural cells during in vivo brain imaging. Nat Neurosci 20:1023-1032|
|Hill, Robert A; Tong, Lei; Yuan, Peng et al. (2015) Regional Blood Flow in the Normal and Ischemic Brain Is Controlled by Arteriolar Smooth Muscle Cell Contractility and Not by Capillary Pericytes. Neuron 87:95-110|
|Hill, Robert A; Grutzendler, Jaime (2014) In vivo imaging of oligodendrocytes with sulforhodamine 101. Nat Methods 11:1081-2|
|Hill, Robert A; Patel, Kiran D; Goncalves, Christopher M et al. (2014) Modulation of oligodendrocyte generation during a critical temporal window after NG2 cell division. Nat Neurosci 17:1518-27|