---
_id: '12875'
abstract:
- lang: eng
  text: The superior colliculus (SC) in the mammalian midbrain is essential for multisensory
    integration and is composed of a rich diversity of excitatory and inhibitory neurons
    and glia. However, the developmental principles directing the generation of SC
    cell-type diversity are not understood. Here, we pursued systematic cell lineage
    tracing in silico and in vivo, preserving full spatial information, using genetic
    mosaic analysis with double markers (MADM)-based clonal analysis with single-cell
    sequencing (MADM-CloneSeq). The analysis of clonally related cell lineages revealed
    that radial glial progenitors (RGPs) in SC are exceptionally multipotent. Individual
    resident RGPs have the capacity to produce all excitatory and inhibitory SC neuron
    types, even at the stage of terminal division. While individual clonal units show
    no pre-defined cellular composition, the establishment of appropriate relative
    proportions of distinct neuronal types occurs in a PTEN-dependent manner. Collectively,
    our findings provide an inaugural framework at the single-RGP/-cell level of the
    mammalian SC ontogeny.
acknowledged_ssus:
- _id: Bio
- _id: M-Shop
- _id: LifeSc
- _id: PreCl
acknowledgement: "We thank Liqun Luo for his continued support, for providing essential
  resources for generating Fzd10-CreER mice which were generated in his laboratory,
  and for comments on the manuscript; W. Zhong for providing Nestin-Cre transgenic
  mouse line for this study; A. Heger for mouse colony management; R. Beattie and
  T. Asenov for designing and producing components of acute slice recovery chamber
  for MADM-CloneSeq experiments; and K. Leopold, J. Rodarte and N. Amberg for initial
  experiments, technical support and/or assistance. This study was supported by the
  Scientific Service Units (SSU) of IST Austria through resources provided by the
  Imaging & Optics Facility (IOF), Laboratory Support Facility (LSF), Miba Machine
  Shop, and Pre-clinical Facility (PCF). G.C. received funding from European Commission
  (IST plus postdoctoral fellowship). This work was supported by ISTA institutional\r\nfunds;
  the Austrian Science Fund Special Research Programmes (FWF SFB F78 Neuro Stem Modulation)
  to S.H. "
article_processing_charge: Yes (via OA deal)
article_type: comment
author:
- first_name: Giselle T
  full_name: Cheung, Giselle T
  id: 471195F6-F248-11E8-B48F-1D18A9856A87
  last_name: Cheung
  orcid: 0000-0001-8457-2572
- first_name: Florian
  full_name: Pauler, Florian
  id: 48EA0138-F248-11E8-B48F-1D18A9856A87
  last_name: Pauler
  orcid: 0000-0002-7462-0048
- first_name: Peter
  full_name: Koppensteiner, Peter
  id: 3B8B25A8-F248-11E8-B48F-1D18A9856A87
  last_name: Koppensteiner
  orcid: 0000-0002-3509-1948
- first_name: Thomas
  full_name: Krausgruber, Thomas
  last_name: Krausgruber
- first_name: Carmen
  full_name: Streicher, Carmen
  id: 36BCB99C-F248-11E8-B48F-1D18A9856A87
  last_name: Streicher
- first_name: Martin
  full_name: Schrammel, Martin
  id: f13e7cae-e8bd-11ed-841a-96dedf69f46d
  last_name: Schrammel
- first_name: Natalie Y
  full_name: Özgen, Natalie Y
  id: e68ece33-f6e0-11ea-865d-ae1031dcc090
  last_name: Özgen
- first_name: Alexis
  full_name: Ivec, Alexis
  id: 1d144691-e8be-11ed-9b33-bdd3077fad4c
  last_name: Ivec
- first_name: Christoph
  full_name: Bock, Christoph
  last_name: Bock
- first_name: Ryuichi
  full_name: Shigemoto, Ryuichi
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
- first_name: Simon
  full_name: Hippenmeyer, Simon
  id: 37B36620-F248-11E8-B48F-1D18A9856A87
  last_name: Hippenmeyer
  orcid: 0000-0003-2279-1061
date_created: 2023-04-27T09:41:48Z
date_published: 2024-01-17T00:00:00Z
date_updated: 2025-05-14T09:39:37Z
day: '17'
ddc:
- '570'
department:
- _id: SiHi
- _id: RySh
doi: 10.1016/j.neuron.2023.11.009
external_id:
  pmid:
  - '38096816'
file:
- access_level: open_access
  checksum: 32b3788f7085cf44a84108d8faaff3ce
  content_type: application/pdf
  creator: dernst
  date_created: 2024-02-06T13:56:15Z
  date_updated: 2024-02-06T13:56:15Z
  file_id: '14944'
  file_name: 2024_Neuron_Cheung.pdf
  file_size: 5942467
  relation: main_file
  success: 1
file_date_updated: 2024-02-06T13:56:15Z
has_accepted_license: '1'
intvolume: '       112'
issue: '2'
language:
- iso: eng
month: '01'
oa: 1
oa_version: Published Version
page: 230-246.e11
pmid: 1
project:
- _id: 059F6AB4-7A3F-11EA-A408-12923DDC885E
  grant_number: F07805
  name: Molecular Mechanisms of Neural Stem Cell Lineage Progression
publication: Neuron
publication_identifier:
  eisbn:
  - '1234995621'
  issn:
  - 0896-6273
  issnl:
  - 1234-5678
publication_status: published
publisher: Elsevier
quality_controlled: '1'
related_material:
  link:
  - description: News on ISTA Website
    relation: press_release
    url: https://ista.ac.at/en/news/the-pedigree-of-brain-cells/
scopus_import: '1'
status: public
title: Multipotent progenitors instruct ontogeny of the superior colliculus
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 112
year: '2024'
...
---
_id: '14683'
abstract:
- lang: eng
  text: "Mosaic analysis with double markers (MADM) technology enables the generation
    of genetic mosaic tissue in mice and high-resolution phenotyping at the individual
    cell level. Here, we present a protocol for isolating MADM-labeled cells with
    high yield for downstream molecular analyses using fluorescence-activated cell
    sorting (FACS). We describe steps for generating MADM-labeled mice, perfusion,
    single-cell suspension, and debris removal. We then detail procedures for cell
    sorting by FACS and downstream analysis. This protocol is suitable for embryonic
    to adult mice.\r\nFor complete details on the use and execution of this protocol,
    please refer to Contreras et al. (2021).1"
acknowledged_ssus:
- _id: Bio
- _id: PreCl
acknowledgement: This research was supported by the Scientific Service Units (SSU)
  at IST Austria through resources provided by the Imaging & Optics Facility (IOF)
  and Preclinical Facilities (PCF). N.A. received support from FWF Firnberg-Programme
  (T 1031). G.C. received support from the European Union’s Horizon 2020 research
  and innovation programme under the Marie Skłodowska-Curie grant agreement no. 754411
  as an ISTplus postdoctoral fellow. This work was also supported by IST Austria institutional
  funds, FWF SFB F78 to S.H., and the European Research Council (ERC) under the European
  Union’s Horizon 2020 research and innovation programme (grant agreement no. 725780
  LinPro) to S.H.
article_number: '102771'
article_processing_charge: No
article_type: review
author:
- first_name: Nicole
  full_name: Amberg, Nicole
  id: 4CD6AAC6-F248-11E8-B48F-1D18A9856A87
  last_name: Amberg
  orcid: 0000-0002-3183-8207
- first_name: Giselle T
  full_name: Cheung, Giselle T
  id: 471195F6-F248-11E8-B48F-1D18A9856A87
  last_name: Cheung
  orcid: 0000-0001-8457-2572
- first_name: Simon
  full_name: Hippenmeyer, Simon
  id: 37B36620-F248-11E8-B48F-1D18A9856A87
  last_name: Hippenmeyer
  orcid: 0000-0003-2279-1061
citation:
  ama: Amberg N, Cheung GT, Hippenmeyer S. Protocol for sorting cells from mouse brains
    labeled with mosaic analysis with double markers by flow cytometry. <i>STAR Protocols</i>.
    2023;5(1). doi:<a href="https://doi.org/10.1016/j.xpro.2023.102771">10.1016/j.xpro.2023.102771</a>
  apa: Amberg, N., Cheung, G. T., &#38; Hippenmeyer, S. (2023). Protocol for sorting
    cells from mouse brains labeled with mosaic analysis with double markers by flow
    cytometry. <i>STAR Protocols</i>. Elsevier. <a href="https://doi.org/10.1016/j.xpro.2023.102771">https://doi.org/10.1016/j.xpro.2023.102771</a>
  chicago: Amberg, Nicole, Giselle T Cheung, and Simon Hippenmeyer. “Protocol for
    Sorting Cells from Mouse Brains Labeled with Mosaic Analysis with Double Markers
    by Flow Cytometry.” <i>STAR Protocols</i>. Elsevier, 2023. <a href="https://doi.org/10.1016/j.xpro.2023.102771">https://doi.org/10.1016/j.xpro.2023.102771</a>.
  ieee: N. Amberg, G. T. Cheung, and S. Hippenmeyer, “Protocol for sorting cells from
    mouse brains labeled with mosaic analysis with double markers by flow cytometry,”
    <i>STAR Protocols</i>, vol. 5, no. 1. Elsevier, 2023.
  ista: Amberg N, Cheung GT, Hippenmeyer S. 2023. Protocol for sorting cells from
    mouse brains labeled with mosaic analysis with double markers by flow cytometry.
    STAR Protocols. 5(1), 102771.
  mla: Amberg, Nicole, et al. “Protocol for Sorting Cells from Mouse Brains Labeled
    with Mosaic Analysis with Double Markers by Flow Cytometry.” <i>STAR Protocols</i>,
    vol. 5, no. 1, 102771, Elsevier, 2023, doi:<a href="https://doi.org/10.1016/j.xpro.2023.102771">10.1016/j.xpro.2023.102771</a>.
  short: N. Amberg, G.T. Cheung, S. Hippenmeyer, STAR Protocols 5 (2023).
date_created: 2023-12-13T11:48:05Z
date_published: 2023-12-08T00:00:00Z
date_updated: 2023-12-18T08:06:14Z
day: '08'
ddc:
- '570'
department:
- _id: SiHi
doi: 10.1016/j.xpro.2023.102771
ec_funded: 1
external_id:
  pmid:
  - '38070137'
intvolume: '         5'
issue: '1'
keyword:
- General Immunology and Microbiology
- General Biochemistry
- Genetics and Molecular Biology
- General Neuroscience
language:
- iso: eng
main_file_link:
- open_access: '1'
  url: https://doi.org/10.1016/j.xpro.2023.102771
month: '12'
oa: 1
oa_version: Submitted Version
pmid: 1
project:
- _id: 268F8446-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: T0101031
  name: Role of Eed in neural stem cell lineage progression
- _id: 260C2330-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '754411'
  name: ISTplus - Postdoctoral Fellowships
- _id: 059F6AB4-7A3F-11EA-A408-12923DDC885E
  grant_number: F07805
  name: Molecular Mechanisms of Neural Stem Cell Lineage Progression
- _id: 260018B0-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '725780'
  name: Principles of Neural Stem Cell Lineage Progression in Cerebral Cortex Development
publication: STAR Protocols
publication_identifier:
  issn:
  - 2666-1667
publication_status: epub_ahead
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: Protocol for sorting cells from mouse brains labeled with mosaic analysis with
  double markers by flow cytometry
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 5
year: '2023'
...
---
_id: '14783'
abstract:
- lang: eng
  text: Connexin 43, an astroglial gap junction protein, is enriched in perisynaptic
    astroglial processes and plays major roles in synaptic transmission. We have previously
    found that astroglial Cx43 controls synaptic glutamate levels and allows for activity-dependent
    glutamine release to sustain physiological synaptic transmissions and cognitiogns.
    However, whether Cx43 is important for the release of synaptic vesicles, which
    is a critical component of synaptic efficacy, remains unanswered. Here, using
    transgenic mice with a glial conditional knockout of Cx43 (Cx43−/−), we investigate
    whether and how astrocytes regulate the release of synaptic vesicles from hippocampal
    synapses. We report that CA1 pyramidal neurons and their synapses develop normally
    in the absence of astroglial Cx43. However, a significant impairment in synaptic
    vesicle distribution and release dynamics were observed. In particular, the FM1-43
    assays performed using two-photon live imaging and combined with multi-electrode
    array stimulation in acute hippocampal slices, revealed a slower rate of synaptic
    vesicle release in Cx43−/− mice. Furthermore, paired-pulse recordings showed that
    synaptic vesicle release probability was also reduced and is dependent on glutamine
    supply via Cx43 hemichannel (HC). Taken together, we have uncovered a role for
    Cx43 in regulating presynaptic functions by controlling the rate and probability
    of synaptic vesicle release. Our findings further highlight the significance of
    astroglial Cx43 in synaptic transmission and efficacy.
acknowledgement: 'This research was funded by grants from the European Research Council
  (Consolidator grant #683154) and European Union’s Horizon 2020 research and innovation
  program (Marie Sklodowska-Curie Innovative Training Networks, grant #722053, EU-GliaPhD)
  to N.R., as well as from FP7-PEOPLE Marie Curie Intra-European Fellowship for career
  development (grant #622289) to G.C. We thank Elena Dossi, Grégory Ghézali, and Jérémie
  Teillon for support with setting up the MEA system for the two-photon microscope.
  We would also like to thank Tayfun Palaz for their technical assistance with the
  EM preparations.'
article_number: '1133'
article_processing_charge: Yes
article_type: original
author:
- first_name: Giselle T
  full_name: Cheung, Giselle T
  id: 471195F6-F248-11E8-B48F-1D18A9856A87
  last_name: Cheung
  orcid: 0000-0001-8457-2572
- first_name: Oana
  full_name: Chever, Oana
  last_name: Chever
- first_name: Astrid
  full_name: Rollenhagen, Astrid
  last_name: Rollenhagen
- first_name: Nicole
  full_name: Quenech’du, Nicole
  last_name: Quenech’du
- first_name: Pascal
  full_name: Ezan, Pascal
  last_name: Ezan
- first_name: Joachim H. R.
  full_name: Lübke, Joachim H. R.
  last_name: Lübke
- first_name: Nathalie
  full_name: Rouach, Nathalie
  last_name: Rouach
citation:
  ama: Cheung GT, Chever O, Rollenhagen A, et al. Astroglial connexin 43 regulates
    synaptic vesicle release at hippocampal synapses. <i>Cells</i>. 2023;12(8). doi:<a
    href="https://doi.org/10.3390/cells12081133">10.3390/cells12081133</a>
  apa: Cheung, G. T., Chever, O., Rollenhagen, A., Quenech’du, N., Ezan, P., Lübke,
    J. H. R., &#38; Rouach, N. (2023). Astroglial connexin 43 regulates synaptic vesicle
    release at hippocampal synapses. <i>Cells</i>. MDPI. <a href="https://doi.org/10.3390/cells12081133">https://doi.org/10.3390/cells12081133</a>
  chicago: Cheung, Giselle T, Oana Chever, Astrid Rollenhagen, Nicole Quenech’du,
    Pascal Ezan, Joachim H. R. Lübke, and Nathalie Rouach. “Astroglial Connexin 43
    Regulates Synaptic Vesicle Release at Hippocampal Synapses.” <i>Cells</i>. MDPI,
    2023. <a href="https://doi.org/10.3390/cells12081133">https://doi.org/10.3390/cells12081133</a>.
  ieee: G. T. Cheung <i>et al.</i>, “Astroglial connexin 43 regulates synaptic vesicle
    release at hippocampal synapses,” <i>Cells</i>, vol. 12, no. 8. MDPI, 2023.
  ista: Cheung GT, Chever O, Rollenhagen A, Quenech’du N, Ezan P, Lübke JHR, Rouach
    N. 2023. Astroglial connexin 43 regulates synaptic vesicle release at hippocampal
    synapses. Cells. 12(8), 1133.
  mla: Cheung, Giselle T., et al. “Astroglial Connexin 43 Regulates Synaptic Vesicle
    Release at Hippocampal Synapses.” <i>Cells</i>, vol. 12, no. 8, 1133, MDPI, 2023,
    doi:<a href="https://doi.org/10.3390/cells12081133">10.3390/cells12081133</a>.
  short: G.T. Cheung, O. Chever, A. Rollenhagen, N. Quenech’du, P. Ezan, J.H.R. Lübke,
    N. Rouach, Cells 12 (2023).
date_created: 2024-01-10T09:46:35Z
date_published: 2023-04-11T00:00:00Z
date_updated: 2024-01-16T09:29:35Z
day: '11'
ddc:
- '570'
department:
- _id: SiHi
doi: 10.3390/cells12081133
external_id:
  isi:
  - '000977445700001'
  pmid:
  - '37190042'
file:
- access_level: open_access
  checksum: 6798cd75d8857976fbc58a43fd173d68
  content_type: application/pdf
  creator: dernst
  date_created: 2024-01-16T09:26:52Z
  date_updated: 2024-01-16T09:26:52Z
  file_id: '14808'
  file_name: 2023_Cells_Cheung.pdf
  file_size: 7931643
  relation: main_file
  success: 1
file_date_updated: 2024-01-16T09:26:52Z
has_accepted_license: '1'
intvolume: '        12'
isi: 1
issue: '8'
keyword:
- General Medicine
language:
- iso: eng
month: '04'
oa: 1
oa_version: Published Version
pmid: 1
publication: Cells
publication_identifier:
  issn:
  - 2073-4409
publication_status: published
publisher: MDPI
quality_controlled: '1'
status: public
title: Astroglial connexin 43 regulates synaptic vesicle release at hippocampal synapses
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 12
year: '2023'
...
---
_id: '10764'
abstract:
- lang: eng
  text: Presynaptic glutamate replenishment is fundamental to brain function. In high
    activity regimes, such as epileptic episodes, this process is thought to rely
    on the glutamate-glutamine cycle between neurons and astrocytes. However the presence
    of an astroglial glutamine supply, as well as its functional relevance in vivo
    in the healthy brain remain controversial, partly due to a lack of tools that
    can directly examine glutamine transfer. Here, we generated a fluorescent probe
    that tracks glutamine in live cells, which provides direct visual evidence of
    an activity-dependent glutamine supply from astroglial networks to presynaptic
    structures under physiological conditions. This mobilization is mediated by connexin43,
    an astroglial protein with both gap-junction and hemichannel functions, and is
    essential for synaptic transmission and object recognition memory. Our findings
    uncover an indispensable recruitment of astroglial glutamine in physiological
    synaptic activity and memory via an unconventional pathway, thus providing an
    astrocyte basis for cognitive processes.
acknowledgement: 'We thank D. Mazaud and. J. Cazères for technical assistance. This
  work was supported by grants from the European Research Council (Consolidator grant
  #683154) and European Union’s Horizon 2020 research and innovation program (Marie
  Sklodowska-Curie Innovative Training Networks, grant #722053, EU-GliaPhD) to N.R.
  and from FP7-PEOPLE Marie Curie Intra-European Fellowship for career development
  (grant #622289) to G.C.'
article_number: '753'
article_processing_charge: No
article_type: original
author:
- first_name: Giselle T
  full_name: Cheung, Giselle T
  id: 471195F6-F248-11E8-B48F-1D18A9856A87
  last_name: Cheung
- first_name: Danijela
  full_name: Bataveljic, Danijela
  last_name: Bataveljic
- first_name: Josien
  full_name: Visser, Josien
  last_name: Visser
- first_name: Naresh
  full_name: Kumar, Naresh
  last_name: Kumar
- first_name: Julien
  full_name: Moulard, Julien
  last_name: Moulard
- first_name: Glenn
  full_name: Dallérac, Glenn
  last_name: Dallérac
- first_name: Daria
  full_name: Mozheiko, Daria
  last_name: Mozheiko
- first_name: Astrid
  full_name: Rollenhagen, Astrid
  last_name: Rollenhagen
- first_name: Pascal
  full_name: Ezan, Pascal
  last_name: Ezan
- first_name: Cédric
  full_name: Mongin, Cédric
  last_name: Mongin
- first_name: Oana
  full_name: Chever, Oana
  last_name: Chever
- first_name: Alexis Pierre
  full_name: Bemelmans, Alexis Pierre
  last_name: Bemelmans
- first_name: Joachim
  full_name: Lübke, Joachim
  last_name: Lübke
- first_name: Isabelle
  full_name: Leray, Isabelle
  last_name: Leray
- first_name: Nathalie
  full_name: Rouach, Nathalie
  last_name: Rouach
citation:
  ama: Cheung GT, Bataveljic D, Visser J, et al. Physiological synaptic activity and
    recognition memory require astroglial glutamine. <i>Nature Communications</i>.
    2022;13. doi:<a href="https://doi.org/10.1038/s41467-022-28331-7">10.1038/s41467-022-28331-7</a>
  apa: Cheung, G. T., Bataveljic, D., Visser, J., Kumar, N., Moulard, J., Dallérac,
    G., … Rouach, N. (2022). Physiological synaptic activity and recognition memory
    require astroglial glutamine. <i>Nature Communications</i>. Springer Nature. <a
    href="https://doi.org/10.1038/s41467-022-28331-7">https://doi.org/10.1038/s41467-022-28331-7</a>
  chicago: Cheung, Giselle T, Danijela Bataveljic, Josien Visser, Naresh Kumar, Julien
    Moulard, Glenn Dallérac, Daria Mozheiko, et al. “Physiological Synaptic Activity
    and Recognition Memory Require Astroglial Glutamine.” <i>Nature Communications</i>.
    Springer Nature, 2022. <a href="https://doi.org/10.1038/s41467-022-28331-7">https://doi.org/10.1038/s41467-022-28331-7</a>.
  ieee: G. T. Cheung <i>et al.</i>, “Physiological synaptic activity and recognition
    memory require astroglial glutamine,” <i>Nature Communications</i>, vol. 13. Springer
    Nature, 2022.
  ista: Cheung GT, Bataveljic D, Visser J, Kumar N, Moulard J, Dallérac G, Mozheiko
    D, Rollenhagen A, Ezan P, Mongin C, Chever O, Bemelmans AP, Lübke J, Leray I,
    Rouach N. 2022. Physiological synaptic activity and recognition memory require
    astroglial glutamine. Nature Communications. 13, 753.
  mla: Cheung, Giselle T., et al. “Physiological Synaptic Activity and Recognition
    Memory Require Astroglial Glutamine.” <i>Nature Communications</i>, vol. 13, 753,
    Springer Nature, 2022, doi:<a href="https://doi.org/10.1038/s41467-022-28331-7">10.1038/s41467-022-28331-7</a>.
  short: G.T. Cheung, D. Bataveljic, J. Visser, N. Kumar, J. Moulard, G. Dallérac,
    D. Mozheiko, A. Rollenhagen, P. Ezan, C. Mongin, O. Chever, A.P. Bemelmans, J.
    Lübke, I. Leray, N. Rouach, Nature Communications 13 (2022).
date_created: 2022-02-20T23:01:30Z
date_published: 2022-02-08T00:00:00Z
date_updated: 2023-08-02T14:25:01Z
day: '08'
ddc:
- '570'
department:
- _id: SiHi
doi: 10.1038/s41467-022-28331-7
external_id:
  isi:
  - '000757297200017'
  pmid:
  - '35136061'
file:
- access_level: open_access
  checksum: 51d580aff2327dd957946208a9749e1a
  content_type: application/pdf
  creator: dernst
  date_created: 2022-02-21T07:51:33Z
  date_updated: 2022-02-21T07:51:33Z
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  file_name: 2022_NatureCommunications_Cheung.pdf
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file_date_updated: 2022-02-21T07:51:33Z
has_accepted_license: '1'
intvolume: '        13'
isi: 1
language:
- iso: eng
month: '02'
oa: 1
oa_version: Published Version
pmid: 1
publication: Nature Communications
publication_identifier:
  eissn:
  - '20411723'
publication_status: published
publisher: Springer Nature
quality_controlled: '1'
scopus_import: '1'
status: public
title: Physiological synaptic activity and recognition memory require astroglial glutamine
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 13
year: '2022'
...
---
_id: '7815'
abstract:
- lang: eng
  text: Beginning from a limited pool of progenitors, the mammalian cerebral cortex
    forms highly organized functional neural circuits. However, the underlying cellular
    and molecular mechanisms regulating lineage transitions of neural stem cells (NSCs)
    and eventual production of neurons and glia in the developing neuroepithelium
    remains unclear. Methods to trace NSC division patterns and map the lineage of
    clonally related cells have advanced dramatically. However, many contemporary
    lineage tracing techniques suffer from the lack of cellular resolution of progeny
    cell fate, which is essential for deciphering progenitor cell division patterns.
    Presented is a protocol using mosaic analysis with double markers (MADM) to perform
    in vivo clonal analysis. MADM concomitantly manipulates individual progenitor
    cells and visualizes precise division patterns and lineage progression at unprecedented
    single cell resolution. MADM-based interchromosomal recombination events during
    the G2-X phase of mitosis, together with temporally inducible CreERT2, provide
    exact information on the birth dates of clones and their division patterns. Thus,
    MADM lineage tracing provides unprecedented qualitative and quantitative optical
    readouts of the proliferation mode of stem cell progenitors at the single cell
    level. MADM also allows for examination of the mechanisms and functional requirements
    of candidate genes in NSC lineage progression. This method is unique in that comparative
    analysis of control and mutant subclones can be performed in the same tissue environment
    in vivo. Here, the protocol is described in detail, and experimental paradigms
    to employ MADM for clonal analysis and lineage tracing in the developing cerebral
    cortex are demonstrated. Importantly, this protocol can be adapted to perform
    MADM clonal analysis in any murine stem cell niche, as long as the CreERT2 driver
    is present.
acknowledged_ssus:
- _id: Bio
- _id: LifeSc
- _id: PreCl
article_number: e61147
article_processing_charge: No
article_type: original
author:
- first_name: Robert J
  full_name: Beattie, Robert J
  id: 2E26DF60-F248-11E8-B48F-1D18A9856A87
  last_name: Beattie
  orcid: 0000-0002-8483-8753
- first_name: Carmen
  full_name: Streicher, Carmen
  id: 36BCB99C-F248-11E8-B48F-1D18A9856A87
  last_name: Streicher
- first_name: Nicole
  full_name: Amberg, Nicole
  id: 4CD6AAC6-F248-11E8-B48F-1D18A9856A87
  last_name: Amberg
  orcid: 0000-0002-3183-8207
- first_name: Giselle T
  full_name: Cheung, Giselle T
  id: 471195F6-F248-11E8-B48F-1D18A9856A87
  last_name: Cheung
  orcid: 0000-0001-8457-2572
- first_name: Ximena
  full_name: Contreras, Ximena
  id: 475990FE-F248-11E8-B48F-1D18A9856A87
  last_name: Contreras
- first_name: Andi H
  full_name: Hansen, Andi H
  id: 38853E16-F248-11E8-B48F-1D18A9856A87
  last_name: Hansen
- first_name: Simon
  full_name: Hippenmeyer, Simon
  id: 37B36620-F248-11E8-B48F-1D18A9856A87
  last_name: Hippenmeyer
  orcid: 0000-0003-2279-1061
citation:
  ama: Beattie RJ, Streicher C, Amberg N, et al. Lineage tracing and clonal analysis
    in developing cerebral cortex using mosaic analysis with double markers (MADM).
    <i>Journal of Visual Experiments</i>. 2020;(159). doi:<a href="https://doi.org/10.3791/61147">10.3791/61147</a>
  apa: Beattie, R. J., Streicher, C., Amberg, N., Cheung, G. T., Contreras, X., Hansen,
    A. H., &#38; Hippenmeyer, S. (2020). Lineage tracing and clonal analysis in developing
    cerebral cortex using mosaic analysis with double markers (MADM). <i>Journal of
    Visual Experiments</i>. MyJove Corporation. <a href="https://doi.org/10.3791/61147">https://doi.org/10.3791/61147</a>
  chicago: Beattie, Robert J, Carmen Streicher, Nicole Amberg, Giselle T Cheung, Ximena
    Contreras, Andi H Hansen, and Simon Hippenmeyer. “Lineage Tracing and Clonal Analysis
    in Developing Cerebral Cortex Using Mosaic Analysis with Double Markers (MADM).”
    <i>Journal of Visual Experiments</i>. MyJove Corporation, 2020. <a href="https://doi.org/10.3791/61147">https://doi.org/10.3791/61147</a>.
  ieee: R. J. Beattie <i>et al.</i>, “Lineage tracing and clonal analysis in developing
    cerebral cortex using mosaic analysis with double markers (MADM),” <i>Journal
    of Visual Experiments</i>, no. 159. MyJove Corporation, 2020.
  ista: Beattie RJ, Streicher C, Amberg N, Cheung GT, Contreras X, Hansen AH, Hippenmeyer
    S. 2020. Lineage tracing and clonal analysis in developing cerebral cortex using
    mosaic analysis with double markers (MADM). Journal of Visual Experiments. (159),
    e61147.
  mla: Beattie, Robert J., et al. “Lineage Tracing and Clonal Analysis in Developing
    Cerebral Cortex Using Mosaic Analysis with Double Markers (MADM).” <i>Journal
    of Visual Experiments</i>, no. 159, e61147, MyJove Corporation, 2020, doi:<a href="https://doi.org/10.3791/61147">10.3791/61147</a>.
  short: R.J. Beattie, C. Streicher, N. Amberg, G.T. Cheung, X. Contreras, A.H. Hansen,
    S. Hippenmeyer, Journal of Visual Experiments (2020).
date_created: 2020-05-11T08:31:20Z
date_published: 2020-05-08T00:00:00Z
date_updated: 2024-03-25T23:30:23Z
day: '08'
ddc:
- '570'
department:
- _id: SiHi
doi: 10.3791/61147
ec_funded: 1
external_id:
  isi:
  - '000546406600043'
file:
- access_level: open_access
  checksum: 3154ea7f90b9fb45e084cd1c2770597d
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  creator: rbeattie
  date_created: 2020-05-11T08:28:38Z
  date_updated: 2020-07-14T12:48:03Z
  file_id: '7816'
  file_name: jove-protocol-61147-lineage-tracing-clonal-analysis-developing-cerebral-cortex-using.pdf
  file_size: 1352186
  relation: main_file
file_date_updated: 2020-07-14T12:48:03Z
has_accepted_license: '1'
isi: 1
issue: '159'
language:
- iso: eng
month: '05'
oa: 1
oa_version: Published Version
project:
- _id: 264E56E2-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: M02416
  name: Molecular Mechanisms Regulating Gliogenesis in the Cerebral Cortex
- _id: 268F8446-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: T0101031
  name: Role of Eed in neural stem cell lineage progression
- _id: 260C2330-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '754411'
  name: ISTplus - Postdoctoral Fellowships
- _id: 2625A13E-B435-11E9-9278-68D0E5697425
  grant_number: '24812'
  name: Molecular Mechanisms of Radial Neuronal Migration
- _id: 260018B0-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '725780'
  name: Principles of Neural Stem Cell Lineage Progression in Cerebral Cortex Development
publication: Journal of Visual Experiments
publication_identifier:
  issn:
  - 1940-087X
publication_status: published
publisher: MyJove Corporation
quality_controlled: '1'
related_material:
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    relation: part_of_dissertation
    status: public
scopus_import: '1'
status: public
title: Lineage tracing and clonal analysis in developing cerebral cortex using mosaic
  analysis with double markers (MADM)
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
year: '2020'
...
---
_id: '7005'
abstract:
- lang: eng
  text: Activity-dependent bulk endocytosis generates synaptic vesicles (SVs) during
    intense neuronal activity via a two-step process. First, bulk endosomes are formed
    direct from the plasma membrane from which SVs are then generated. SV generation
    from bulk endosomes requires the efflux of previously accumulated calcium and
    activation of the protein phosphatase calcineurin. However, it is still unknown
    how calcineurin mediates SV generation. We addressed this question using a series
    of acute interventions that decoupled the generation of SVs from bulk endosomes
    in rat primary neuronal culture. This was achieved by either disruption of protein–protein
    interactions via delivery of competitive peptides, or inhibition of enzyme activity
    by known inhibitors. SV generation was monitored using either a morphological
    horseradish peroxidase assay or an optical assay that monitors the replenishment
    of the reserve SV pool. We found that SV generation was inhibited by, (i) peptides
    that disrupt calcineurin interactions, (ii) an inhibitor of dynamin I GTPase activity
    and (iii) peptides that disrupt the phosphorylation-dependent dynamin I–syndapin
    I interaction. Peptides that disrupted syndapin I interactions with eps15 homology
    domain-containing proteins had no effect. This revealed that (i) calcineurin must
    be localized at bulk endosomes to mediate its effect, (ii) dynamin I GTPase activity
    is essential for SV fission and (iii) the calcineurin-dependent interaction between
    dynamin I and syndapin I is essential for SV generation. We therefore propose
    that a calcineurin-dependent dephosphorylation cascade that requires both dynamin
    I GTPase and syndapin I lipid-deforming activity is essential for SV generation
    from bulk endosomes.
article_processing_charge: No
article_type: original
author:
- first_name: Giselle T
  full_name: Cheung, Giselle T
  id: 471195F6-F248-11E8-B48F-1D18A9856A87
  last_name: Cheung
  orcid: 0000-0001-8457-2572
- first_name: Michael A.
  full_name: Cousin, Michael A.
  last_name: Cousin
citation:
  ama: Cheung GT, Cousin MA. Synaptic vesicle generation from activity‐dependent bulk
    endosomes requires a dephosphorylation‐dependent dynamin–syndapin interaction.
    <i>Journal of Neurochemistry</i>. 2019;151(5):570-583. doi:<a href="https://doi.org/10.1111/jnc.14862">10.1111/jnc.14862</a>
  apa: Cheung, G. T., &#38; Cousin, M. A. (2019). Synaptic vesicle generation from
    activity‐dependent bulk endosomes requires a dephosphorylation‐dependent dynamin–syndapin
    interaction. <i>Journal of Neurochemistry</i>. Wiley. <a href="https://doi.org/10.1111/jnc.14862">https://doi.org/10.1111/jnc.14862</a>
  chicago: Cheung, Giselle T, and Michael A. Cousin. “Synaptic Vesicle Generation
    from Activity‐dependent Bulk Endosomes Requires a Dephosphorylation‐dependent
    Dynamin–Syndapin Interaction.” <i>Journal of Neurochemistry</i>. Wiley, 2019.
    <a href="https://doi.org/10.1111/jnc.14862">https://doi.org/10.1111/jnc.14862</a>.
  ieee: G. T. Cheung and M. A. Cousin, “Synaptic vesicle generation from activity‐dependent
    bulk endosomes requires a dephosphorylation‐dependent dynamin–syndapin interaction,”
    <i>Journal of Neurochemistry</i>, vol. 151, no. 5. Wiley, pp. 570–583, 2019.
  ista: Cheung GT, Cousin MA. 2019. Synaptic vesicle generation from activity‐dependent
    bulk endosomes requires a dephosphorylation‐dependent dynamin–syndapin interaction.
    Journal of Neurochemistry. 151(5), 570–583.
  mla: Cheung, Giselle T., and Michael A. Cousin. “Synaptic Vesicle Generation from
    Activity‐dependent Bulk Endosomes Requires a Dephosphorylation‐dependent Dynamin–Syndapin
    Interaction.” <i>Journal of Neurochemistry</i>, vol. 151, no. 5, Wiley, 2019,
    pp. 570–83, doi:<a href="https://doi.org/10.1111/jnc.14862">10.1111/jnc.14862</a>.
  short: G.T. Cheung, M.A. Cousin, Journal of Neurochemistry 151 (2019) 570–583.
date_created: 2019-11-12T14:37:08Z
date_published: 2019-12-01T00:00:00Z
date_updated: 2023-08-30T07:21:50Z
day: '01'
ddc:
- '570'
department:
- _id: SiHi
doi: 10.1111/jnc.14862
external_id:
  isi:
  - '000490703100001'
  pmid:
  - '31479508'
file:
- access_level: open_access
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  content_type: application/pdf
  creator: dernst
  date_created: 2020-02-05T10:30:02Z
  date_updated: 2020-07-14T12:47:47Z
  file_id: '7452'
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  file_size: 4334962
  relation: main_file
file_date_updated: 2020-07-14T12:47:47Z
has_accepted_license: '1'
intvolume: '       151'
isi: 1
issue: '5'
language:
- iso: eng
month: '12'
oa: 1
oa_version: Published Version
page: 570-583
pmid: 1
publication: Journal of Neurochemistry
publication_identifier:
  eissn:
  - 1471-4159
  issn:
  - 0022-3042
publication_status: published
publisher: Wiley
quality_controlled: '1'
scopus_import: '1'
status: public
title: Synaptic vesicle generation from activity‐dependent bulk endosomes requires
  a dephosphorylation‐dependent dynamin–syndapin interaction
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 151
year: '2019'
...