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Neuroscience

쥐 뇌 조직 검사 정렬 형광 활성 셀을 사용하여 셀 타입 별 유전자 발현

Published: May 28, 2015 doi: 10.3791/52537

Materials

Name Company Catalog Number Comments
Neural Dissociation Kit (P) Miltenyi Biotec 130-092-628
Myelin Removal Beads II Miltenyi Biotec 130-096-733
LS Columns Miltenyi Biotec 130-042-401
QuadroMACS Separator Miltenyi Biotec 130-090-976
MACS MultiStand Miltenyi Biotec 130-042-303
Nylon Mesh Sheet Amazon CMN-0074-10YD 40 inch width, 80 micron size mesh
Fc Block / anti-CD32 BD Biosciences BDB550270 reactivity for rat
APC-conjugated CD11b antibody Biolegend 201809 reactivity for rat
Rabbit anti-GLT1 Novus Biologicals NBP1-20136 reactivity for rat or human
PE-conjugated anti-rabbit secondary antibody eBioscience 1037259 secondary antibody for anti-GLT1
FITC-conjugated anti-rat CD90 (Thy1) mouse antibody Biolegend 202504 reactivity for rat

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References

  1. Schwarz, J. M., Smith, S. H., Bilbo, S. D. FACS analysis of neuronal-glial interactions in the nucleus accumbens following morphine administration. Psychopharmacology. 230 (4), 525-535 (2013).
  2. Schwarz, J. M., Hutchinson, M. R., Bilbo, S. D. Early-life experience decreases drug-induced reinstatement of morphine CPP in adulthood via microglial-specific epigenetic programming of anti-inflammatory IL-10 expression. J Neurosci. 31 (49), 17835-1523 (2011).
  3. Herzenberg, L. A., Tung, J., Moore, W. A., Herzenberg, L. A., Parks, D. R. Interpreting flow cytometry data: a guide for the perplexed. Nature Immunology. 7 (7), 681-685 (2006).
  4. Guez-Barber, D., et al. FACS identifies unique cocaine-induced gene regulation in selectively activated adult striatal neurons. J Neurosci. 31 (11), 4251-4259 (2011).
  5. Fanous, S., et al. Unique gene alterations are induced in FACS-purified Fos-positive neurons activated during cue-induced relapse to heroin seeking. J Neurochem. 124 (1), 100-108 (2013).
  6. Liu, Q. R., et al. Detection of molecular alterations in methamphetamine-activated Fos-expressing neurons from single rat dorsal striatum using fluorescence-activated cell sorting (FACS). J Neurochem. 128 (1), 173-185 (2013).
  7. Guez-Barber, D., et al. FACS purification of immunolabeled cell types from adult rat brain. J Neurosci Methods. 203 (1), 10-18 (2012).
  8. Nolte, C., et al. GFAP promoter-controlled EGFP-expressing transgenic mice: a tool to visualize astrocytes and astrogliosis in living brain tissue. Glia. 33 (1), 72-86 (2000).
  9. Okana, M., Bell, D. W., Haber, D. A., Li, E. DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell. 99 (3), 247-257 (1999).
  10. Bogdanović, O., Veenstra, G. J. DNA methylation and methyl-CpG binding proteins: developmental requirements and function. Chromosoma. 118 (5), 549-565 (2009).
  11. Iwamoto, K., et al. Neurons show distinctive DNA methylation profile and higher inter-individual variations compared with non-neurons. Genome Res. 21 (5), 688-696 (2011).
  12. Nishioka, M., et al. Neuronal cell-type specific DNA methylation patterns of the Cacna1c gene. Int J Dev Neurosci. 31 (2), 89-95 (2013).
  13. Kozlenkov, A., et al. Differences in DNA methylation between human neuronal and glial cells are concentrated in enhancers and non-CpG sites. Nucleic Acid Res. 42 (1), 109-127 (2014).
  14. Russo, S. J., et al. Nuclear factor kappa B signaling regulates neuronal morphology and cocaine reward. J Neurosci. 29 (11), 3529-3537 (2009).
  15. Bhatt, D., Ghosh, S. Regulation of the NF-κB-Mediated Transcription of Inflammatory Genes. Front Immunol. 5, 71 (2014).
  16. Okada, S., et al. Flow cytometric sorting of neuronal and glial nuclei from central nervous system tissue. J Cell Physiol. 226 (2), 552-558 (2011).
쥐 뇌 조직 검사 정렬 형광 활성 셀을 사용하여 셀 타입 별 유전자 발현
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Cite this Article

Schwarz, J. M. Using FluorescenceMore

Schwarz, J. M. Using Fluorescence Activated Cell Sorting to Examine Cell-Type-Specific Gene Expression in Rat Brain Tissue. J. Vis. Exp. (99), e52537, doi:10.3791/52537 (2015).

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