Establishing an adult mouse brain hippocampal organotypic slice culture system that allows for tracing and pharmacological manipulation of ex vivo neurogenesis

Mayerl, Steffen and ffrench-Constant, Charles ORCID: https://orcid.org/0000-0002-5621-3377 (2021) Establishing an adult mouse brain hippocampal organotypic slice culture system that allows for tracing and pharmacological manipulation of ex vivo neurogenesis. BIO-Protocol, 11 (1). ISSN 2331-8325

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Abstract

The function of the hippocampus depends on the process of adult hippocampal neurogenesis which underpins the exceptional neural plasticity of this structure, and is also frequently affected in CNS pathologies. Thus, manipulation of this process represents an important therapeutic goal. To identify potential strategies, organotypic adult brain slices are emerging as a valuable tool. Over the recent years, this methodology has been refined and here we present a combined protocol that brings together these refinements to enable long-term culture of adult hippocampal slices. We employ a sectioning technique that retains essential afferent inputs onto the hippocampus as well as serum-free culture conditions, so allowing an extended culture period. To sustain the neurogenic potential in the slices, we utilize the gliogenesis-inhibitor Indomethacin. Using EdU retention analysis enables us to assess the effects of pharmacological intervention on neurogenesis. With these improvements, we have established an easy and reliable method to study the effects of small molecules/drugs on proliferation and neuron formation ex vivo which will facilitate future discovery driven drug screenings.

Item Type: Article
Additional Information: Funding Information: This work was supported by a grant from the DFG to SM (MA7212/ 2-1), by a Wellcome Trust Senior Investigator award (104783/Z/14/Z) and funding from the BBSRC (BB/L00402X/1) to CffC. We thank SCRM animal facility staff (Luke McPhee, Chris Wilson, Lorraine McNeil, John Agnew, and Jamie Kelly) and SCRM imaging facility staff (Bertrand Vernay) for their excellent help. The protocol was first published in our original research paper “Hippocampal Neurogenesis Requires Cell-Autonomous Thyroid Hormone Signaling” in Stem Cell Reports (Mayerl et al., 2020).
Uncontrolled Keywords: adult hippocampal neurogenesis,drug screening,indomethacin,mct8,organotypic adult slice culture,thyroid hormone,biochemistry, genetics and molecular biology(all),immunology and microbiology(all),neuroscience(all),plant science ,/dk/atira/pure/subjectarea/asjc/1300
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
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Depositing User: LivePure Connector
Date Deposited: 20 Jul 2022 10:30
Last Modified: 12 Aug 2022 05:36
URI: https://ueaeprints.uea.ac.uk/id/eprint/86713
DOI: 10.21769/BioProtoc.3869

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