---
_id: '7406'
abstract:
- lang: eng
  text: "Background\r\nSynaptic vesicles (SVs) are an integral part of the neurotransmission
    machinery, and isolation of SVs from their host neuron is necessary to reveal
    their most fundamental biochemical and functional properties in in vitro assays.
    Isolated SVs from neurons that have been genetically engineered, e.g. to introduce
    genetically encoded indicators, are not readily available but would permit new
    insights into SV structure and function. Furthermore, it is unclear if cultured
    neurons can provide sufficient starting material for SV isolation procedures.\r\n\r\nNew
    method\r\nHere, we demonstrate an efficient ex vivo procedure to obtain functional
    SVs from cultured rat cortical neurons after genetic engineering with a lentivirus.\r\n\r\nResults\r\nWe
    show that ∼108 plated cortical neurons allow isolation of suitable SV amounts
    for functional analysis and imaging. We found that SVs isolated from cultured
    neurons have neurotransmitter uptake comparable to that of SVs isolated from intact
    cortex. Using total internal reflection fluorescence (TIRF) microscopy, we visualized
    an exogenous SV-targeted marker protein and demonstrated the high efficiency of
    SV modification.\r\n\r\nComparison with existing methods\r\nObtaining SVs from
    genetically engineered neurons currently generally requires the availability of
    transgenic animals, which is constrained by technical (e.g. cost and time) and
    biological (e.g. developmental defects and lethality) limitations.\r\n\r\nConclusions\r\nThese
    results demonstrate the modification and isolation of functional SVs using cultured
    neurons and viral transduction. The ability to readily obtain SVs from genetically
    engineered neurons will permit linking in situ studies to in vitro experiments
    in a variety of genetic contexts."
acknowledged_ssus:
- _id: Bio
- _id: EM-Fac
article_processing_charge: No
article_type: original
author:
- first_name: Catherine
  full_name: Mckenzie, Catherine
  id: 3EEDE19A-F248-11E8-B48F-1D18A9856A87
  last_name: Mckenzie
- first_name: Miroslava
  full_name: Spanova, Miroslava
  id: 44A924DC-F248-11E8-B48F-1D18A9856A87
  last_name: Spanova
- first_name: Alexander J
  full_name: Johnson, Alexander J
  id: 46A62C3A-F248-11E8-B48F-1D18A9856A87
  last_name: Johnson
  orcid: 0000-0002-2739-8843
- first_name: Stephanie
  full_name: Kainrath, Stephanie
  id: 32CFBA64-F248-11E8-B48F-1D18A9856A87
  last_name: Kainrath
- first_name: Vanessa
  full_name: Zheden, Vanessa
  id: 39C5A68A-F248-11E8-B48F-1D18A9856A87
  last_name: Zheden
  orcid: 0000-0002-9438-4783
- first_name: Harald H.
  full_name: Sitte, Harald H.
  last_name: Sitte
- first_name: Harald L
  full_name: Janovjak, Harald L
  id: 33BA6C30-F248-11E8-B48F-1D18A9856A87
  last_name: Janovjak
  orcid: 0000-0002-8023-9315
citation:
  ama: Mckenzie C, Spanova M, Johnson AJ, et al. Isolation of synaptic vesicles from
    genetically engineered cultured neurons. <i>Journal of Neuroscience Methods</i>.
    2019;312:114-121. doi:<a href="https://doi.org/10.1016/j.jneumeth.2018.11.018">10.1016/j.jneumeth.2018.11.018</a>
  apa: Mckenzie, C., Spanova, M., Johnson, A. J., Kainrath, S., Zheden, V., Sitte,
    H. H., &#38; Janovjak, H. L. (2019). Isolation of synaptic vesicles from genetically
    engineered cultured neurons. <i>Journal of Neuroscience Methods</i>. Elsevier.
    <a href="https://doi.org/10.1016/j.jneumeth.2018.11.018">https://doi.org/10.1016/j.jneumeth.2018.11.018</a>
  chicago: Mckenzie, Catherine, Miroslava Spanova, Alexander J Johnson, Stephanie
    Kainrath, Vanessa Zheden, Harald H. Sitte, and Harald L Janovjak. “Isolation of
    Synaptic Vesicles from Genetically Engineered Cultured Neurons.” <i>Journal of
    Neuroscience Methods</i>. Elsevier, 2019. <a href="https://doi.org/10.1016/j.jneumeth.2018.11.018">https://doi.org/10.1016/j.jneumeth.2018.11.018</a>.
  ieee: C. Mckenzie <i>et al.</i>, “Isolation of synaptic vesicles from genetically
    engineered cultured neurons,” <i>Journal of Neuroscience Methods</i>, vol. 312.
    Elsevier, pp. 114–121, 2019.
  ista: Mckenzie C, Spanova M, Johnson AJ, Kainrath S, Zheden V, Sitte HH, Janovjak
    HL. 2019. Isolation of synaptic vesicles from genetically engineered cultured
    neurons. Journal of Neuroscience Methods. 312, 114–121.
  mla: Mckenzie, Catherine, et al. “Isolation of Synaptic Vesicles from Genetically
    Engineered Cultured Neurons.” <i>Journal of Neuroscience Methods</i>, vol. 312,
    Elsevier, 2019, pp. 114–21, doi:<a href="https://doi.org/10.1016/j.jneumeth.2018.11.018">10.1016/j.jneumeth.2018.11.018</a>.
  short: C. Mckenzie, M. Spanova, A.J. Johnson, S. Kainrath, V. Zheden, H.H. Sitte,
    H.L. Janovjak, Journal of Neuroscience Methods 312 (2019) 114–121.
date_created: 2020-01-30T09:12:19Z
date_published: 2019-01-15T00:00:00Z
date_updated: 2023-09-06T15:27:29Z
day: '15'
department:
- _id: HaJa
- _id: Bio
doi: 10.1016/j.jneumeth.2018.11.018
ec_funded: 1
external_id:
  isi:
  - '000456220900013'
  pmid:
  - '30496761'
intvolume: '       312'
isi: 1
language:
- iso: eng
month: '01'
oa_version: None
page: 114-121
pmid: 1
project:
- _id: 25548C20-B435-11E9-9278-68D0E5697425
  call_identifier: FP7
  grant_number: '303564'
  name: Microbial Ion Channels for Synthetic Neurobiology
- _id: 26538374-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: I03630
  name: Molecular mechanisms of endocytic cargo recognition in plants
- _id: 2548AE96-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: W1232-B24
  name: Molecular Drug Targets
publication: Journal of Neuroscience Methods
publication_identifier:
  issn:
  - 0165-0270
publication_status: published
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: Isolation of synaptic vesicles from genetically engineered cultured neurons
type: journal_article
user_id: c635000d-4b10-11ee-a964-aac5a93f6ac1
volume: 312
year: '2019'
...