The Journal of Neuroscience, August 13, 2008, 28(33):8294-8305; doi:10.1523/JNEUROSCI.2010-08.2008
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Cellular/Molecular
A Novel Purification Method for CNS Projection Neurons Leads to the Identification of Brain Vascular Cells as a Source of Trophic Support for Corticospinal Motor Neurons
Jason C. Dugas, *
Wim Mandemakers, *
Madolyn Rogers,
Adiljan Ibrahim,
Richard Daneman, and
Ben A. Barres
Department of Neurobiology, Stanford University School of Medicine, Stanford, California 94305-5125
Correspondence should be addressed to Dr. Jason C. Dugas, Department of Neurobiology, Stanford University School of Medicine, Fairchild Building, Room D205, 299 Campus Drive, Stanford, CA 94305-5125. Email: jcdugas{at}alum.mit.edu
One of the difficulties in studying cellular interactions in the CNS is the lack of effective methods to purify specific neuronal populations of interest. We report the development of a novel purification scheme, cholera toxin β (CTB) immunopanning, in which a particular CNS neuron population is selectively labeled via retrograde axonal transport of the cell-surface epitope CTB, and then purified via immobilization with anti-CTB antibody. We have demonstrated the usefulness and versatility of this method by purifying both retinal ganglion cells and corticospinal motor neurons (CSMNs). Genomic expression analyses of purified CSMNs revealed that they express significant levels of many receptors for growth factors produced by brain endothelial cells; three of these factors, CXCL12, pleiotrophin, and IGF2 significantly enhanced purified CSMN survival, similar to previously characterized CSMN trophic factors BDNF and IGF1. In addition, endothelial cell conditioned medium significantly promoted CSMN neurite outgrowth. These findings demonstrate a useful method for the purification of several different types of CNS projection neurons, which in principle should work in many mammalian species, and provide evidence that endothelial-derived factors may represent an overlooked source of trophic support for neurons in the brain.
Key words: corticospinal motoneurons; immunopanning; endothelial cells; IGF2; CXCL12; SDF1; pleiotrophin
Received May 1, 2008;
revised June 23, 2008;
accepted June 30, 2008.
Correspondence should be addressed to Dr. Jason C. Dugas, Department of Neurobiology, Stanford University School of Medicine, Fairchild Building, Room D205, 299 Campus Drive, Stanford, CA 94305-5125. Email: jcdugas{at}alum.mit.edu
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K. Arai and E. H. Lo
An Oligovascular Niche: Cerebral Endothelial Cells Promote the Survival and Proliferation of Oligodendrocyte Precursor Cells
J. Neurosci.,
April 8, 2009;
29(14):
4351 - 4355.
[Abstract]
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