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
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Schmitz C, Hof PR (2005) Design-based stereology in neuroscience. Neuroscience 130:813–831
Sterio DC (1984) The unbiased estimation of number and sizes of arbitrary particles using the disector. J Microsc 134:127–136
Schweichel JU, Merker HJ (1973) The morphology of various types of cell death in prenatal tissues. Teratology 7:253–266
Schmued LC, Hopkins KJ (2000) Fluoro-Jade B: a high affinity fluorescent marker for the localization of neuronal degeneration. Brain Res 874:123–130
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
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
Hirsch EC, Hunot S (2009) Neuroinflammation in Parkinson’s disease: a target for neuroprotection? Lancet Neurol 8:382–397
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
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
Tenner AJ (2001) Complement in Alzheimer’s disease: opportunities for modulating protective and pathogenic events. Neurobiol Aging 22:849–861
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
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
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
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
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
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
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
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
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
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
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
Editor information
Editors and Affiliations
Rights 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