Skip to main content
SpringerLink
Log in

Preparation of Rat Hippocampal Organotypic Cultures and Application to Study Amyloid β-Peptide Toxicity

  • Protocol
  • First Online:
Book cover Neurotrophic Factors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1727))

  • 3399 Accesses

Abstract

Hippocampal organotypic cultures constitute a very easy but delicate method widely used to study amyloid β-peptide toxicity. This ex vivo technique is performed on tissues isolated from newborn rats. Here, we describe a protocol for the preparation and culture of hippocampal organotypic slices that can be maintained for 14–21 days and their application to the study of amyloid β-peptide toxicity.

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

Access this chapter

Log in via an institution

Institutional subscriptions

References

  1. Reinbold R (1954) Organotypic differentiation of the eye of the chick embryo in vitro. C R Seances Soc Biol Fil 148:1493–1495

    CAS  PubMed  Google Scholar 

  2. Scuderi C, Valenza M, Stecca C et al (2012) Palmitoylethanolamide exerts neuroprotective effects in mixed neuroglial cultures and organotypic hippocampal slices via peroxisome proliferator-activated receptor-α. J Neuroinflammation 9:49. https://doi.org/10.1186/1742-2094-9-21

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Harwell CS, Coleman MP (2016) Synaptophysin depletion and intraneuronal Aβ in organotypic hippocampal slice cultures from huAPP transgenic mice. Mol Neurodegener 11:44. https://doi.org/10.1186/s13024-016-0110-7

    Article  PubMed  PubMed Central  Google Scholar 

  4. Müller M, Gähwiler BH, Rietschin L et al (1993) Reversible loss of dendritic spines and altered excitability after chronic epilepsy in hippocampal slice cultures. Proc Natl Acad Sci U S A 90:257–261

    Article  PubMed  PubMed Central  Google Scholar 

  5. Gähwiler BH, Capogna M, Debanne D et al (1997) Organotypic slice cultures: a technique has come of age. Trends Neurosci 20:471–477

    Article  PubMed  Google Scholar 

  6. Stoppini L, Buchs PA, Muller D (1991) A simple method for organotypic cultures of nervous tissue. J Neurosci Methods 37:173–182

    Article  CAS  PubMed  Google Scholar 

  7. Shamir ER, Ewald AJ (2014) Three-dimensional organotypic culture: experimental models of mammalian biology and disease. Nat Rev Mol Cell Biol 15:647–664. https://doi.org/10.1038/nrm3873

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Ullrich C, Daschil N, Humpel C (2011) Organotypic vibrosections: novel whole sagittal brain cultures. J Neurosci Methods 201:131–141. https://doi.org/10.1016/j.jneumeth.2011.07.021

    Article  PubMed  PubMed Central  Google Scholar 

  9. Pellegrini-Giampietro DE, Cozzi A, Peruginelli F et al (1999) 1-Aminoindan-1,5-dicarboxylic acid and (S)-(+)-2-(3′-carboxybicyclo[1,1,1]pentyl)-glycine, two mGlu1 receptor-preferring antagonists, reduce neuronal death in in vitro and in vivo models of cerebral ischemia. Eur J Neurosci 11:3637–3647. https://doi.org/10.1046/j.1460-9568.1999.00786.x

    Article  CAS  PubMed  Google Scholar 

  10. Finley M, Fairman D, Liu D et al (2004) Functional validation of adult hippocampal organotypic cultures as an in vitro model of brain injury. Brain Res 100:125–132

    Article  Google Scholar 

  11. Scuderi C, Steardo L (2013) Neuroglial roots of neurodegenerative diseases: therapeutic potential of palmitoylethanolamide in models of Alzheimer’s disease. CNS Neurol Disord Drug Targets 12:62–69

    Article  CAS  PubMed  Google Scholar 

  12. Hardy J, Selkoe DJ (2002) The amyloid hypothesis of Alzheimer’s disease: progress and problems on the road to therapeutics. Science 297:353–356

    Article  CAS  PubMed  Google Scholar 

  13. Opitz-Araya X, Barria A (2011) Organotypic hippocampal slice cultures. J Vis Exp 48:2462. https://doi.org/10.3791/2462

    Google Scholar 

  14. Stine WB, Jungbauer L, Yu C, LaDu MJ (2011) Preparing synthetic Aβ in different aggregation states. Methods Mol Biol 670:13–32. https://doi.org/10.1007/978-1-60761-744-0_2

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Paxinos G, Watson C (1998) The rat brain in stereotaxic coordinates, 4th De Luxe edn. Elsevier Science Publishing Co Inc., San Diego

    Google Scholar 

  16. Franklin KBJ, Paxinos G (2001) The mouse brain in stereotaxic coordinates, 2nd revised edn. Elsevier Science Publishing Co Inc, San Diego

    Google Scholar 

Download references

Acknowledgement

This work was supported by SAPIENZA University Grant to CS (prot. C26A15X58E).

Author information

Authors and Affiliations

  1. Department of Physiology and Pharmacology “V. Erspamer”, SAPIENZA University of Rome, Rome, Italy

    Maria Rosanna Bronzuoli, Roberta Facchinetti & Caterina Scuderi

Authors
  1. Maria Rosanna Bronzuoli
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Roberta Facchinetti
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Caterina Scuderi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Caterina Scuderi .

Editor information

Editors and Affiliations

  1. Department of Pharmaceutical and Pharmacological Sciences, University of Padova, Padova, Italy

    Stephen D. Skaper

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Bronzuoli, M.R., Facchinetti, R., Scuderi, C. (2018). Preparation of Rat Hippocampal Organotypic Cultures and Application to Study Amyloid β-Peptide Toxicity. In: Skaper, S. (eds) Neurotrophic Factors. Methods in Molecular Biology, vol 1727. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7571-6_24

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7571-6_24

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7570-9

  • Online ISBN: 978-1-4939-7571-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation