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  Single-cell axotomy of cultured hippocampal neurons integrated in neuronal circuits

Gomis-Rüth, S., Stiess, M., Wierenga, C. J., Meyn, L., & Bradke, F. (2014). Single-cell axotomy of cultured hippocampal neurons integrated in neuronal circuits. Nature Protocols, 9(5), 1028-1037. doi:10.1038/nprot.2014.069.

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Gomis-Rüth, Susana1, Author              
Stiess, Michael1, Author              
Wierenga, Corette J., Author
Meyn, Liane1, Author              
Bradke, Frank1, Author              
Affiliations:
1Max Planck Research Group: Axonal Growth and Regeneration / Bradke, MPI of Neurobiology, Max Planck Society, ou_1113553              

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Free keywords: SPINAL-CORD REGENERATION; AXONAL REGENERATION; IN-VIVO; STRETCH INJURY; GROWTH CONE; POLARITY; DENDRITES; PLASTICITY; MODELS; VITRO
 Abstract: An understanding of the molecular mechanisms of axon regeneration after injury is key for the development of potential therapies. Single-cell axotomy of dissociated neurons enables the study of the intrinsic regenerative capacities of injured axons. This protocol describes how to perform single-cell axotomy on dissociated hippocampal neurons containing synapses. Furthermore, to axotomize hippocampal neurons integrated in neuronal circuits, we describe how to set up coculture with a few fluorescently labeled neurons. This approach allows axotomy of single cells in a complex neuronal network and the observation of morphological and molecular changes during axon regeneration. Thus, single-cell axotomy of mature neurons is a valuable tool for gaining insights into cell intrinsic axon regeneration and the plasticity of neuronal polarity of mature neurons. Dissociation of the hippocampus and plating of hippocampal neurons takes similar to 2 h. Neurons are then left to grow for 2 weeks, during which time they integrate into neuronal circuits. Subsequent axotomy takes 10 min per neuron and further imaging takes 10 min per neuron.

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Language(s): eng - English
 Dates: 2014-05
 Publication Status: Published in print
 Pages: 10
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: ISI: 000335014500003
DOI: 10.1038/nprot.2014.069
 Degree: -

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Title: Nature Protocols
  Other : Nat. Protoc.
Source Genre: Journal
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Publ. Info: London, UK : Nature Publishing Group
Pages: - Volume / Issue: 9 (5) Sequence Number: - Start / End Page: 1028 - 1037 Identifier: ISSN: 1750-2799
CoNE: https://pure.mpg.de/cone/journals/resource/1000000000223800_1