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Laser Capture Microdissection in Traumatic Brain Injury Research: Obtaining Hippocampal Subregions and Pools of Injured Neurons for Genomic Analyses

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Laser Capture Microdissection

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1723))

Abstract

The methods presented here are based on our laboratory’s 15 years of experience using laser capture microdissection to obtain samples for the study of gene expression after traumatic brain injury (TBI) using a well-established rat model of experimental TBI. Here, we describe how to use the ArcturusXT laser capture microdissection system to capture swaths of specific regions of the rat hippocampus as well as specific neuronal populations defined by Fluoro-Jade C staining. Staining with Fluoro-Jade C identifies a neuron that is in the process of degeneration. We have optimized our protocols for Fluoro-Jade C tissue staining and laser capture microdissection to maintain RNA integrity which is essential for a variety of downstream applications, such as microarray, PCR array, and quantitative real-time PCR analyses.

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References

  1. Emmert-Buck MR, Bonner RF, Smith PD et al (1996) Laser capture microdissection. Science 274:998–1001

    Article  CAS  Google Scholar 

  2. Bota M, Dong HW, Swanson LW (2003) From gene networks to brain networks. Nat Neurosci 6:795–799

    Article  CAS  Google Scholar 

  3. Boone DR, Sell SL, Hellmich HL (2012) Laser capture microdissection of enriched populations of neurons or single neurons for gene expression analysis after traumatic brain injury. J Vis Exp 74:1–7

    Google Scholar 

  4. Shimamura M, Garcia JM, Prough DS, Hellmich HL (2004) Laser capture microdissection and analysis of amplified antisense RNA from distinct cell populations of the young and aged rat brain: effect of traumatic brain injury on hippocampal gene expression. Mol Brain Res 17:47–61

    Article  Google Scholar 

  5. Shah SA, Prough DS, Garcia JM, DeWitt DS, Hellmich HL (2006) Molecular correlates of age-specific responses to traumatic brain injury in mice. Exp Gerontol 41(11):1201–1205. Epub 2006 Sep 15

    Article  CAS  Google Scholar 

  6. Kiraly M, Kiraly SJ (2007) Traumatic brain injury and delayed sequelae: a review—traumatic brain injury and mild traumatic brain injury (Concussion) are precursors to later-onset brain disorders, including early-onset dementia. ScientificWorldJournal 7:1768–1776

    Article  Google Scholar 

  7. Blennow K, Hardy J, Zetterberg H (2012) The neuropathology and neurobiology of traumatic brain injury. Neuron 76:886–899

    Article  CAS  Google Scholar 

  8. Boone DR, Sell SL, Micci MA et al (2012) Traumatic brain injury-induced dysregulation of the circadian clock. PLoS One 7:e46204

    Article  CAS  Google Scholar 

  9. Rojo DR, Prough DS, Boone DR et al (2011) Influence of stochastic gene expression on the cell survival rheostat after traumatic brain injury. PLoS One 6:e23111

    Article  CAS  Google Scholar 

  10. Hellmich HL, Rojo DR, Micci MA et al (2013) Pathway analysis reveals common pro-survival mechanisms of metyrapone and carbenoxolone after traumatic brain injury. PLoS One 8:e53230

    Article  CAS  Google Scholar 

  11. Boone DR, Micci MA, Taglialatela IG et al (2015) Pathway-focused PCR array profiling of enriched populations of laser capture microdissected hippocampal cells after traumatic brain injury. PLoS One 10:e0127287

    Article  Google Scholar 

  12. Datta S, Malhotra L, Dickerson R, Chaffee S, Sen CK, Roy S (2015) Laser capture microdissection: Big data from small samples. Histol Histopathol 30:1255–1269

    CAS  Google Scholar 

  13. Kruse CS, Goswamy R, Raval Y, Marawi S (2016) Challenges and opportunities of big data in health care: a systematic review. JMIR Med Inform 4:e38–v4i4e38

    Article  Google Scholar 

  14. Kohli MA, Lucae S, Saemann PG et al (2011) The neuronal transporter gene SLC6A15 confers risk to major depression. Neuron 70:252–265

    Article  CAS  Google Scholar 

  15. Dwivedi Y (2013) microRNAs as biomarker in depression pathogeneisis. Ann Psychiatry Mental Health 1:1–5

    Google Scholar 

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Acknowledgments

This work was supported in part by an NIH grant (RO1 NS052532) to HLH, by the Department of Anesthesiology, UTMB Galveston, and by the Moody Project for Translational Traumatic Brain Injury Research. We thank Christy Perry for assistance with the illustrations.

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Authors and Affiliations

  1. Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA

    Deborah R. Boone, Harris A. Weisz, Stacy L. Sell & Helen L. Hellmich

Authors
  1. Deborah R. Boone
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  2. Harris A. Weisz
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  3. Stacy L. Sell
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  4. Helen L. Hellmich
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Corresponding author

Correspondence to Helen L. Hellmich .

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Editors and Affiliations

  1. School of Medicine, Medical Sciences and Nutrition, University of Aberdeen, Aberdeen, United Kingdom

    Graeme I. Murray

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Boone, D.R., Weisz, H.A., Sell, S.L., Hellmich, H.L. (2018). Laser Capture Microdissection in Traumatic Brain Injury Research: Obtaining Hippocampal Subregions and Pools of Injured Neurons for Genomic Analyses. In: Murray, G. (eds) Laser Capture Microdissection. Methods in Molecular Biology, vol 1723. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7558-7_13

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  • DOI: https://doi.org/10.1007/978-1-4939-7558-7_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7557-0

  • Online ISBN: 978-1-4939-7558-7

  • eBook Packages: Springer Protocols

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