Skip to main content
Book cover

Necrosis pp 91–113Cite as

Histological Analysis of Neurodegeneration in the Mouse Brain

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1004))

Abstract

Neurodegenerative diseases such as Alzheimer’s disease (AD) and Parkinson’s disease (PD) are characterized by chronic and progressive neuronal loss. Being able to detect and quantify neurodegeneration is the first step to identify mechanisms underlying neuronal cell death and to develop novel therapeutic strategies. In this chapter, we describe a practical method for detecting and quantifying neurodegeneration in adult and aging mouse brains based on protocols developed in our laboratory over the last decade. We include protocols on sample preparation, immunohistochemical analysis, and stereological methods for counting neurons using examples of AD and PD mouse models. We also describe how to use Fluoro-Jade staining and terminal deoxynucleotidyl transferase dUTP nick end labeling to detect degenerating neurons and apoptotic cells, respectively, and how to use specific proteins as early markers of neurodegeneration.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   79.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   99.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Schmitz C, Hof PR (2005) Design-based ­stereology in neuroscience. Neuroscience 130:813–831

    Article  CAS  Google Scholar 

  2. Sterio DC (1984) The unbiased estimation of number and sizes of arbitrary particles using the disector. J Microsc 134:127–136

    Article  CAS  Google Scholar 

  3. Schweichel JU, Merker HJ (1973) The morphology of various types of cell death in prenatal tissues. Teratology 7:253–266

    Article  CAS  Google Scholar 

  4. Schmued LC, Hopkins KJ (2000) Fluoro-Jade B: a high affinity fluorescent marker for the localization of neuronal degeneration. Brain Res 874:123–130

    Article  CAS  Google Scholar 

  5. Negoescu A, Lorimier P, Labat-Moleur F, Drouet C, Robert C, Guillermet C, Brambilla C, Brambilla E (1996) In situ apoptotic cell labeling by the TUNEL method: improvement and evaluation on cell preparations. J Histochem Cytochem 44:959–968

    Article  CAS  Google Scholar 

  6. McGeer PL, Itagaki S, Boyes BE, McGeer EG (1988) Reactive microglia are positive for HLA-DR in the substantia nigra of Parkinson’s and Alzheimer’s disease brains. Neurology 38:1285–1291

    Article  CAS  Google Scholar 

  7. Hirsch EC, Hunot S (2009) Neuroinflammation in Parkinson’s disease: a target for neuroprotection? Lancet Neurol 8:382–397

    Article  CAS  Google Scholar 

  8. Akiyama H, Barger S, Barnum S, Bradt B, Bauer J, Cole GM, Cooper NR, Eikelenboom P, Emmerling M, Fiebich BL et al (2000) Inflammation and Alzheimer’s disease. Neurobiol Aging 21:383–421

    Article  CAS  Google Scholar 

  9. O’Callaghan JP, Jensen KF (1992) Enhanced expression of glial fibrillary acidic protein and the cupric silver degeneration reaction can be used as sensitive and early indicators of neurotoxicity. Neurotoxicology 13:113–122

    PubMed  Google Scholar 

  10. Tenner AJ (2001) Complement in Alzheimer’s disease: opportunities for modulating protective and pathogenic events. Neurobiol Aging 22:849–861

    Article  CAS  Google Scholar 

  11. Lemere CA, Munger JS, Shi GP, Natkin L, Haass C, Chapman HA, Selkoe DJ (1995) The lysosomal cysteine protease, cathepsin S, is increased in Alzheimer’s disease and Down syndrome brain. An immunocytochemical study. Am J Pathol 146:848–860

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Saura CA, Choi SY, Beglopoulos V, Malkani S, Zhang D, Shankaranarayana Rao BS, Chattarji S, Kelleher RJ 3rd, Kandel ER, Duff K et al (2004) Loss of presenilin function causes impairments of memory and synaptic plasticity followed by age-dependent ­neurodegeneration. Neuron 42:23–36

    Article  CAS  Google Scholar 

  13. Beglopoulos V, Sun X, Saura CA, Lemere CA, Kim RD, Shen J (2004) Reduced beta-amyloid production and increased inflammatory responses in presenilin conditional knock-out mice. J Biol Chem 279:46907–46914

    Article  CAS  Google Scholar 

  14. Wines-Samuelson M, Schulte EC, Smith MJ, Aoki C, Liu X, Kelleher RJ 3rd, Shen J (2010) Characterization of age-dependent and progressive cortical neuronal degeneration in ­presenilin conditional mutant mice. PLoS One 5:e10195

    Article  Google Scholar 

  15. Gundersen HJ (1986) Stereology of arbitrary particles. A review of unbiased number and size estimators and the presentation of some new ones, in memory of William R. Thompson. J Microsc 143:3–45

    Article  CAS  Google Scholar 

  16. Gundersen HJ, Bagger P, Bendtsen TF, Evans SM, Korbo L, Marcussen N, Moller A, Nielsen K, Nyengaard JR, Pakkenberg B et al (1988) The new stereological tools: disector, ­fractionator, nucleator and point sampled intercepts and their use in pathological research and diagnosis. APMIS 96:857–881

    Article  CAS  Google Scholar 

  17. West MJ, Slomianka L, Gundersen HJ (1991) Unbiased stereological estimation of the total number of neurons in the subdivisions of the rat hippocampus using the optical fractionator. Anat Rec 231:482–497

    Article  CAS  Google Scholar 

  18. Ward TS, Rosen GD, von Bartheld CS (2008) Optical disector counting in cryosections and vibratome sections underestimates particle numbers: effects of tissue quality. Microsc Res Tech 71:60–68

    Article  Google Scholar 

  19. Evans SM, Nyengaard JR (2004) Number: section introduction. In: Evans SM, Janson AM, Nyengaard JR (eds) Quantitative methods in neuroscience. Oxford University Press, Oxford, pp 61–89

    Chapter  Google Scholar 

  20. Romijn HJ, van Uum JF, Breedijk I, Emmering J, Radu I, Pool CW (1999) Double immunolabeling of neuropeptides in the human hypothalamus as analyzed by confocal laser scanning fluorescence microscopy. J Histochem Cytochem 47:229–236

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We thank Carlos A. Saura, Vassilios Beglopoulos, and Mary Wines-Samuelson for the images in Fig. 2. We thank all current and former members of the Shen lab for the protocols and discussions.

Funding: This work was supported by grants from the National Institutes of Health (R01NS041779, R01NS041783, R01NS042818, R01NS071251).

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Yamaguchi, H., Shen, J. (2013). Histological Analysis of Neurodegeneration in the Mouse Brain. In: McCall, K., Klein, C. (eds) Necrosis. Methods in Molecular Biology, vol 1004. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-383-1_8

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-383-1_8

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-382-4

  • Online ISBN: 978-1-62703-383-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics