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

Overview

Journal J Neurosci

Specialty Neurology

Date 2008 Aug 15

PMID 18701692

Citations 60

Authors

Jason C Dugas

Wim Mandemakers

Madolyn Rogers

Adiljan Ibrahim

Richard Daneman

Ben A Barres

Affiliations

Soon will be listed here.

Abstract

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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References

Herculano-Houzel S, Mota B, Lent R . Cellular scaling rules for rodent brains. Proc Natl Acad Sci U S A. 2006; 103(32):12138-43. PMC: 1567708. DOI: 10.1073/pnas.0604911103. View

Doherty P, Walsh F . Ganglioside GM1 antibodies and B-cholera toxin bind specifically to embryonic chick dorsal root ganglion neurons but do not modulate neurite regeneration. J Neurochem. 1987; 48(4):1237-44. DOI: 10.1111/j.1471-4159.1987.tb05652.x. View

Young R . Cell death during differentiation of the retina in the mouse. J Comp Neurol. 1984; 229(3):362-73. DOI: 10.1002/cne.902290307. View

Spreafico R, Frassoni C, Arcelli P, Selvaggio M, De Biasi S . In situ labeling of apoptotic cell death in the cerebral cortex and thalamus of rats during development. J Comp Neurol. 1995; 363(2):281-95. DOI: 10.1002/cne.903630209. View

Molyneaux B, Arlotta P, Menezes J, Macklis J . Neuronal subtype specification in the cerebral cortex. Nat Rev Neurosci. 2007; 8(6):427-37. DOI: 10.1038/nrn2151. View

Nedelkoska L, Benjamins J . Binding of cholera toxin B subunit: a surface marker for murine microglia but not oligodendrocytes or astrocytes. J Neurosci Res. 1998; 53(5):605-12. DOI: 10.1002/(SICI)1097-4547(19980901)53:5<605::AID-JNR10>3.0.CO;2-#. View

Shen Q, Goderie S, Jin L, Karanth N, Sun Y, Abramova N . Endothelial cells stimulate self-renewal and expand neurogenesis of neural stem cells. Science. 2004; 304(5675):1338-40. DOI: 10.1126/science.1095505. View

Junger H, Junger W . CNTF and GDNF, but not NT-4, support corticospinal motor neuron growth via direct mechanisms. Neuroreport. 1998; 9(16):3749-54. DOI: 10.1097/00001756-199811160-00033. View

Chalasani S, Baribaud F, Coughlan C, Sunshine M, Lee V, Doms R . The chemokine stromal cell-derived factor-1 promotes the survival of embryonic retinal ganglion cells. J Neurosci. 2003; 23(11):4601-12. PMC: 6740796. View

10.

Leventhal C, Rafii S, Rafii D, Shahar A, Goldman S . Endothelial trophic support of neuronal production and recruitment from the adult mammalian subependyma. Mol Cell Neurosci. 1999; 13(6):450-64. DOI: 10.1006/mcne.1999.0762. View

11.

Koh S, Kim Y, Kim H, Cho G, Kim K, Kim S . Recombinant human erythropoietin suppresses symptom onset and progression of G93A-SOD1 mouse model of ALS by preventing motor neuron death and inflammation. Eur J Neurosci. 2007; 25(7):1923-30. DOI: 10.1111/j.1460-9568.2007.05471.x. View

12.

Giehl K, Rohrig S, Bonatz H, Gutjahr M, Leiner B, Bartke I . Endogenous brain-derived neurotrophic factor and neurotrophin-3 antagonistically regulate survival of axotomized corticospinal neurons in vivo. J Neurosci. 2001; 21(10):3492-502. PMC: 2710112. View

13.

Schwab M . Repairing the injured spinal cord. Science. 2002; 295(5557):1029-31. DOI: 10.1126/science.1067840. View

14.

Cuatrecasas P . Gangliosides and membrane receptors for cholera toxin. Biochemistry. 1973; 12(18):3558-66. DOI: 10.1021/bi00742a032. View

15.

Ullian E, Sapperstein S, Christopherson K, Barres B . Control of synapse number by glia. Science. 2001; 291(5504):657-61. DOI: 10.1126/science.291.5504.657. View

16.

Wang Y, Mao X, Xie L, Banwait S, Marti H, Greenberg D . Vascular endothelial growth factor overexpression delays neurodegeneration and prolongs survival in amyotrophic lateral sclerosis mice. J Neurosci. 2007; 27(2):304-7. PMC: 2830908. DOI: 10.1523/JNEUROSCI.4433-06.2007. View

17.

Hammond E, Tetzlaff W, Mestres P, Giehl K . BDNF, but not NT-3, promotes long-term survival of axotomized adult rat corticospinal neurons in vivo. Neuroreport. 1999; 10(12):2671-5. DOI: 10.1097/00001756-199908200-00043. View

18.

Kaur C, Ling E, Wong W . Lectin labelling of amoeboid microglial cells in the brain of postnatal rats. J Anat. 1990; 173:151-60. PMC: 1256090. View

19.

Lu P, Blesch A, Tuszynski M . Neurotrophism without neurotropism: BDNF promotes survival but not growth of lesioned corticospinal neurons. J Comp Neurol. 2001; 436(4):456-70. DOI: 10.1002/cne.1080. View

20.

Herculano-Houzel S, Lent R . Isotropic fractionator: a simple, rapid method for the quantification of total cell and neuron numbers in the brain. J Neurosci. 2005; 25(10):2518-21. PMC: 6725175. DOI: 10.1523/JNEUROSCI.4526-04.2005. View