Journal
JOURNAL OF NEUROSCIENCE
Volume 28, Issue 33, Pages 8294-8305Publisher
SOC NEUROSCIENCE
DOI: 10.1523/JNEUROSCI.2010-08.2008
Keywords
corticospinal motoneurons; immunopanning; endothelial cells; IGF2; CXCL12; SDF1; pleiotrophin
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Funding
- National Eye Institute Regeneration [R01 EY011310]
- Adelson Medical Research Foundation
- National Multiple Sclerosis Society Postdoctoral Fellowship [FG 1434-A- 1]
- Netherlands Organization for Scientific Research TALENT [S 93-380]
- Howard Hughes Medical Institute Predoctoral Fellowship
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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 beta(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.
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