Anthony Brown, Ph.D.
Associate Professor
Department of Neuroscience and
Center for Molecular Neurobiology
Degree: King's College, University of London
Postdoctoral Training: Case Western Reserve University and Temple University
Phone: (614) 292 1205
Fax: (614) 292-5379
Email: brown.2302@osu.edu
Link to NLM PubMed publications list for Tony Brown (last 10 years)
Research Area:
The mechanism of slow axonal transport
Current Research:
Cytoskeletal and cytosolic proteins are synthesized in the cell body of nerve cells and transported out along the axons by slow axonal transport. This movement is essential for the growth and survival of axons and continues throughout the life of the neuron, but the mechanism is not known.
Our most recent studies have focused on the axonal transport of neurofilaments, which are one of three classes of cytoskeletal polymers that comprise the neuronal cytoskeleton. Abnormalities in the axonal transport of neurofilaments are thought to underlie the etiology of a number of neurodegenerative diseases, most notably amyotrophic lateral sclerosis (Lou Gehrig's disease).
We have succeeded in observing the slow axonal transport of GFP-tagged neurofilament protein in cultured nerve cells. Our data indicate that neurofilament proteins move as assembled polymers, an issue that has been the subject of considerable controversy for many years. Contrary to expectations, the neurofilaments move very rapidly, but they also spend most of their time not moving. Based on these observations, we have proposed a "stop and go" model for slow axonal transport in which the slow rate is actually due to rapid movements interrupted by prolonged pauses.
Presently we are using state-of-the-art live-cell fluorescence imaging techniques in combination with molecular, biochemical and ultrastructural approaches to investigate the movement of neurofilaments and other cytoskeletal proteins in cultured nerve cells. Our long-term goal is to define the cargo structures that convey cytoskeletal and cytosolic proteins along axons, the motors that propel them, the substrates along which they move, and the mechanisms that regulate their movement.
Techniques:
Current projects include the use of the following techniques:
Microscopy: epifluorescence, phase contrast and DIC microscopy of living cells; immunofluorescence light microscopy; transmission electron microscopy
Digital imaging: multiple wavelength fluorescence timelapse image acquisition; motion analysis; movie processing; quantification of fluorescence; general digital image processing and analysis
Cell culture: culture of central and peripheral neurons from rats and from wild type and mutant mice; nuclear and cytoplasmic microinjection; transfection with plasmid expression vectors; RNA interference
Biochemistry: purification and covalent modification of cytoskeletal proteins; SDS-PAGE; Western blotting; co-immunoprecipitation assays
Molecular biology: construction of plasmid and viral expression vectors encoding GFP-fusion proteins; site directed mutagenesis
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