---
OA_place: repository
OA_type: green
_id: '21291'
abstract:
- lang: eng
text: The complexity and specificity of movement in vertebrates is driven by a rich
diversity of spinal motor and interneuron cell types. During development, eleven
spinal cord progenitor domains generate an equivalent number of cardinal neuron
types. How progenitor domains, individual progenitors, and post-mitotic diversity
relate is still unknown. We performed high-resolution, single-progenitor cell
lineage tracing in the embryonic mouse spinal cord using mosaic analysis with
double markers (MADM). Our quantitative study of lineage progression revealed
that spinal cord progenitors undergo highly variable numbers of proliferative,
neurogenic, and gliogenic cell divisions. The nascent clonally-related neurons
migrate radially over large distances, span the dorsoventral axis, and even cross
the midline, demonstrating striking bilaterality. Molecular and morphometric analysis
indicate high levels of progenitor multipotency, with an individual progenitor
capable of producing several molecularly and morphologically distinct neuron types,
as well as astrocytes. These findings redefine spinal cord development as a process
in which lineage variability — rather than rigid progenitor identity — drives
the generation of cellular diversity.
acknowledged_ssus:
- _id: PreCl
- _id: Bio
acknowledgement: "We would like to thank Elizabeth Marin, Anna Kicheva, Igor Adameyko,
and James Briscoe as\r\nwell as members of the Sweeney and Hippemeyer labs and SFB
consortium for comments on\r\nthe manuscript. We are also grateful for the technical
support of the Preclinical and Imaging and\r\nOptics Facilities support teams (ISTA).
In addition, we thank our funding sources for providing\r\nthe resources to do these
experiments: Horizon Europe ERC Starting Grant Number 101041551\r\n(M.S.; L.B.S.);
Special Research Program (SFB) of the Austrian Science Fund (FWF)\r\nNeuroStem Modulation
Project numbers F7814-B (S.A.G.; M.S.; G.S.; and L.B.S.) and F7805\r\n(G.C. and
S.H.). S.A.G is supported by a Boehringer Ingelheim Fonds PhD Fellowship, F.D.S.N.\r\nby
an Institute of Science and Technology Austria (ISTA) GROW fellowship, and G.C.
by an\r\nISTA Plus postdoctoral fellowship from the European Commission. S.H./L.B.S.
and G.C. were\r\nadditionally supported by institutional funds from the ISTA and
the University of Exeter,\r\nrespectively. "
article_processing_charge: No
author:
- first_name: Sophie A
full_name: Gobeil, Sophie A
id: 2f3e9efb-eb24-11ec-86b2-88efb11d59fa
last_name: Gobeil
- first_name: Francisco
full_name: Da Silveira Neto, Francisco
id: 8cfb7412-10a7-11f1-add1-82b44e6418f2
last_name: Da Silveira Neto
- first_name: Giulia
full_name: Silvestrelli, Giulia
id: 12632ae8-799e-11ef-94a2-e5a3b5ef49e9
last_name: Silvestrelli
- first_name: Matthijs Geert
full_name: Smits, Matthijs Geert
id: 7a231d52-e216-11ee-a0bb-8acd55f8f1f0
last_name: Smits
- first_name: Carmen
full_name: Streicher, Carmen
id: 36BCB99C-F248-11E8-B48F-1D18A9856A87
last_name: Streicher
- 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
- first_name: Lora Beatrice Jaeger
full_name: Sweeney, Lora Beatrice Jaeger
id: 56BE8254-C4F0-11E9-8E45-0B23E6697425
last_name: Sweeney
orcid: 0000-0001-9242-5601
citation:
ama: Gobeil SA, Da Silveira Neto F, Silvestrelli G, et al. Lineage origin of spinal
cord cell type diversity. bioRxiv. doi:10.64898/2026.02.12.705305
apa: Gobeil, S. A., Da Silveira Neto, F., Silvestrelli, G., Smits, M. G., Streicher,
C., Cheung, G. T., … Sweeney, L. B. (n.d.). Lineage origin of spinal cord cell
type diversity. bioRxiv. https://doi.org/10.64898/2026.02.12.705305
chicago: Gobeil, Sophie A, Francisco Da Silveira Neto, Giulia Silvestrelli, Matthijs
Geert Smits, Carmen Streicher, Giselle T Cheung, Simon Hippenmeyer, and Lora B.
Sweeney. “Lineage Origin of Spinal Cord Cell Type Diversity.” BioRxiv,
n.d. https://doi.org/10.64898/2026.02.12.705305.
ieee: S. A. Gobeil et al., “Lineage origin of spinal cord cell type diversity,”
bioRxiv. .
ista: Gobeil SA, Da Silveira Neto F, Silvestrelli G, Smits MG, Streicher C, Cheung
GT, Hippenmeyer S, Sweeney LB. Lineage origin of spinal cord cell type diversity.
bioRxiv, 10.64898/2026.02.12.705305.
mla: Gobeil, Sophie A., et al. “Lineage Origin of Spinal Cord Cell Type Diversity.”
BioRxiv, doi:10.64898/2026.02.12.705305.
short: S.A. Gobeil, F. Da Silveira Neto, G. Silvestrelli, M.G. Smits, C. Streicher,
G.T. Cheung, S. Hippenmeyer, L.B. Sweeney, BioRxiv (n.d.).
corr_author: '1'
date_created: 2026-02-17T11:36:20Z
date_published: 2026-02-16T00:00:00Z
date_updated: 2026-04-14T08:16:55Z
day: '16'
ddc:
- '570'
department:
- _id: SiHi
- _id: LoSw
doi: 10.64898/2026.02.12.705305
fulldoi: https://doi.org/10.64898/2026.02.12.705305
has_accepted_license: '1'
language:
- iso: eng
license: https://creativecommons.org/licenses/by-nc-nd/4.0/
main_file_link:
- open_access: '1'
url: https://doi.org/10.64898/2026.02.12.705305
month: '02'
oa: 1
oa_version: Preprint
project:
- _id: ebb66355-77a9-11ec-83b8-b8ac210a4dae
grant_number: '101041551'
name: Development and Evolution of Tetrapod Motor Circuits
- _id: 8da85f50-16d5-11f0-9cad-eab8b0ff6c9e
grant_number: F7814
name: 'Stem Cell Modulation in Neural Development and Regeneration/ P14-Swim-to-limb
transition: cell type to connection diversity'
- _id: 059F6AB4-7A3F-11EA-A408-12923DDC885E
grant_number: F7805
name: Stem Cell Modulation in Neural Development and Regeneration/ P05-Molecular
Mechanisms of Neural Stem Cell Lineage Progression
publication: bioRxiv
publication_status: submitted
status: public
title: Lineage origin of spinal cord cell type diversity
tmp:
image: /images/cc_by_nc_nd.png
legal_code_url: https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode
name: Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
(CC BY-NC-ND 4.0)
short: CC BY-NC-ND (4.0)
type: preprint
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
year: '2026'
...
---
OA_place: publisher
OA_type: hybrid
PlanS_conform: '1'
_id: '22753'
abstract:
- lang: eng
text: Radial glial progenitors (RGPs) produce all excitatory neurons in the developing
cerebral cortex. Mosaic analysis with double markers (MADM)-based lineage tracing
in vivo has revealed a quantitative framework of RGP lineage progression1. Here
we established MADM technology2,3 in mouse embryonic stem cells to probe RGP lineage
progression in a self-organizing cortical organoid system. We found that RGPs
exhibit a high level of plasticity in proliferative potential in organoids rather
than strict temporally stereotyped lineage progression as observed in vivo. RGPs
in organoids showed increased lineage restriction, diminishing cell-type diversity
in clones of cortical projection neurons, despite uniform single-cell transcriptional
signatures of RGPs and a unitary lineage trajectory. Thus, critical non-cell-autonomous
cues that are absent in self-organizing systems and/or the genuine stem cell niche
are essential for faithful temporal control of RGP lineage progression and the
generation of clonal cortical cell-type diversity.
acknowledged_ssus:
- _id: Bio
- _id: LifeSc
- _id: PreCl
acknowledgement: We thank M. L. de Guevara, S. Jayaram and A. Heger for technical
assistance with mESC derivation; M. Goudarzi for assistance with organoid imaging;
M. Leeb and F. Freeman for advice in culturing mESCs and organoids; S. Gobeil and
L. Sweeney for reagents and advice for organoid clearing; A. Heger for mouse colony
management; J. Hauser for technical assistance; the Stanford Brain Organogenesis
Workshop; and all members of the Hippenmeyer laboratory for discussion and/or comments
on the manuscript. This study was supported by the Scientific Service Units (SSU)
of the Institute of Science and Technology, Austria through resources provided by
the Imaging and Optics Facility (IOF), Laboratory Support Facility (LSF) and Preclinical
Facility (PCF). M.S. received funding from the European Commission (IST plus postdoctoral
fellowship). This work was supported by ISTA institutional funds to S.H., FWF SFB
F78 Neuro Stem Modulation to S.H., and by the European Research Council (ERC) under
the European Union’s Horizon 2020 Research And Innovation Program (grant agreement
725780 LinPro) to S.H. Open access funding provided by Institute of Science and
Technology (IST Austria).
article_processing_charge: Yes (via OA deal)
article_type: original
author:
- first_name: Melissa A
full_name: Stouffer, Melissa A
id: 4C9372C4-F248-11E8-B48F-1D18A9856A87
last_name: Stouffer
- first_name: Osvaldo
full_name: Miranda, Osvaldo
id: 862A3C56-A8BF-11E9-B4FA-D9E3E5697425
last_name: Miranda
orcid: 0000-0001-6618-6889
- first_name: Florian
full_name: Pauler, Florian
id: 48EA0138-F248-11E8-B48F-1D18A9856A87
last_name: Pauler
orcid: 0000-0002-7462-0048
- first_name: Fabrizia
full_name: Pipicelli, Fabrizia
id: 649134fd-d012-11ed-8f82-db1e5050f9ba
last_name: Pipicelli
- first_name: Carmen
full_name: Streicher, Carmen
id: 36BCB99C-F248-11E8-B48F-1D18A9856A87
last_name: Streicher
- 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: Stouffer MA, Miranda O, Pauler F, et al. Temporal uncoupling of radial glia
lineage progression in cortical organoids. Nature. 2026. doi:10.1038/s41586-026-10916-7
apa: Stouffer, M. A., Miranda, O., Pauler, F., Pipicelli, F., Streicher, C., Cheung,
G. T., & Hippenmeyer, S. (2026). Temporal uncoupling of radial glia lineage
progression in cortical organoids. Nature. Springer Nature. https://doi.org/10.1038/s41586-026-10916-7
chicago: Stouffer, Melissa A, Osvaldo Miranda, Florian Pauler, Fabrizia Pipicelli,
Carmen Streicher, Giselle T Cheung, and Simon Hippenmeyer. “Temporal Uncoupling
of Radial Glia Lineage Progression in Cortical Organoids.” Nature. Springer
Nature, 2026. https://doi.org/10.1038/s41586-026-10916-7.
ieee: M. A. Stouffer et al., “Temporal uncoupling of radial glia lineage
progression in cortical organoids,” Nature. Springer Nature, 2026.
ista: Stouffer MA, Miranda O, Pauler F, Pipicelli F, Streicher C, Cheung GT, Hippenmeyer
S. 2026. Temporal uncoupling of radial glia lineage progression in cortical organoids.
Nature.
mla: Stouffer, Melissa A., et al. “Temporal Uncoupling of Radial Glia Lineage Progression
in Cortical Organoids.” Nature, Springer Nature, 2026, doi:10.1038/s41586-026-10916-7.
short: M.A. Stouffer, O. Miranda, F. Pauler, F. Pipicelli, C. Streicher, G.T. Cheung,
S. Hippenmeyer, Nature (2026).
corr_author: '1'
das_tickbox: '1'
dataavailabilitystatement: All data generated and analysed in this study are included
in the paper, source data and/or Supplementary Tables 2 and 3. Raw sequencing data
have been deposited with Gene Expression Omnibus (GEO) accession number GSE327470.
Source data are provided with this paper. All scripts used to prepare data and figures
for this manuscript are accessible on GitHub at https://github.com/fpauler/Temporal-Uncoupling-of-Radial-Glia-Lineage-Progression-in-Cortical-Organoid.
date_created: 2026-08-23T22:01:47Z
date_published: 2026-08-12T00:00:00Z
date_updated: 2026-09-09T07:11:02Z
day: '12'
ddc:
- '570'
department:
- _id: SiHi
doi: 10.1038/s41586-026-10916-7
ec_funded: 1
external_id:
pmid:
- '42587153'
file:
- access_level: open_access
checksum: 11383e28fc430b28d2666f73833e8b56
content_type: application/pdf
creator: dernst
date_created: 2026-09-07T13:24:10Z
date_updated: 2026-09-07T13:24:10Z
file_id: '22844'
file_name: 2026_Nature_Stouffer.pdf
file_size: 48513068
relation: main_file
success: 1
file_date_updated: 2026-09-07T13:24:10Z
fulldoi: https://doi.org/10.1038/s41586-026-10916-7
has_accepted_license: '1'
language:
- iso: eng
license: https://creativecommons.org/licenses/by/4.0/
month: '08'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: 059F6AB4-7A3F-11EA-A408-12923DDC885E
grant_number: F7805
name: Stem Cell Modulation in Neural Development and Regeneration/ P05-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: Nature
publication_identifier:
eissn:
- 1476-4687
issn:
- 0028-0836
publication_status: epub_ahead
publisher: Springer Nature
quality_controlled: '1'
researchdata_availability: yes
scopus_import: '1'
status: public
supplementarymaterial: yes
title: Temporal uncoupling of radial glia lineage progression in cortical organoids
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: '2026'
...
---
OA_type: closed access
_id: '18765'
abstract:
- lang: eng
text: Mosaic Analysis with Double Markers (MADM) represents a mouse genetic approach
coupling differential fluorescent labeling to genetic manipulations in dividing
cells and their lineages. MADM uniquely enables the generation and visualization
of individual control or homozygous mutant cells in a heterozygous genetic environment.
Among its diverse applications, MADM has been used to dissect cell-autonomous
gene functions important for cortical development and neural development in general.
The high cellular resolution offered by MADM also permits the analysis of transcriptomic
changes of individual cells upon genetic manipulations. In this chapter, we describe
an experimental protocol combining the generation and isolation of MADM-labeled
cells with downstream single-cell RNA-sequencing technologies to probe cell-type
specific phenotypes due to genetic mutations at single-cell resolution.
acknowledged_ssus:
- _id: Bio
acknowledgement: 'We thank all Hippenmeyer lab members for support and discussions.
Experimental steps described were optimized with support provided by the Imaging
& Optics Facility (IOF) and Preclinical Facility (PCF) at ISTA, Vienna BioCenter
Core Facilities (VBCF), and Christoph Bock lab at Center for Molecular Medicine
(CeMM). G.C. received funding from European Commission (IST plus postdoctoral fellowship).
This work was supported by ISTA institutional funds: The Austrian Science Fund Special
Research Programmes (FWF SFB F78 Neuro Stem Modulation) to S.H.'
alternative_title:
- Methods in Molecular Biology
article_processing_charge: No
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: Simon
full_name: Hippenmeyer, Simon
id: 37B36620-F248-11E8-B48F-1D18A9856A87
last_name: Hippenmeyer
orcid: 0000-0003-2279-1061
citation:
ama: 'Cheung GT, Pauler F, Hippenmeyer S. Probing Cell-Type Specificity of Mutant
Phenotype at Transcriptomic Level Using Mosaic Analysis with Double Markers (MADM).
In: Garcia-Marques J, Lee T, eds. Lineage Tracing. Vol 2886. MIMB. New
York, NY: Springer Nature; 2025:139-151. doi:10.1007/978-1-0716-4310-5_7'
apa: 'Cheung, G. T., Pauler, F., & Hippenmeyer, S. (2025). Probing Cell-Type
Specificity of Mutant Phenotype at Transcriptomic Level Using Mosaic Analysis
with Double Markers (MADM). In J. Garcia-Marques & T. Lee (Eds.), Lineage
Tracing (Vol. 2886, pp. 139–151). New York, NY: Springer Nature. https://doi.org/10.1007/978-1-0716-4310-5_7'
chicago: 'Cheung, Giselle T, Florian Pauler, and Simon Hippenmeyer. “Probing Cell-Type
Specificity of Mutant Phenotype at Transcriptomic Level Using Mosaic Analysis
with Double Markers (MADM).” In Lineage Tracing, edited by Jorge Garcia-Marques
and Tzumin Lee, 2886:139–51. MIMB. New York, NY: Springer Nature, 2025. https://doi.org/10.1007/978-1-0716-4310-5_7.'
ieee: 'G. T. Cheung, F. Pauler, and S. Hippenmeyer, “Probing Cell-Type Specificity
of Mutant Phenotype at Transcriptomic Level Using Mosaic Analysis with Double
Markers (MADM),” in Lineage Tracing, vol. 2886, J. Garcia-Marques and T.
Lee, Eds. New York, NY: Springer Nature, 2025, pp. 139–151.'
ista: 'Cheung GT, Pauler F, Hippenmeyer S. 2025.Probing Cell-Type Specificity of
Mutant Phenotype at Transcriptomic Level Using Mosaic Analysis with Double Markers
(MADM). In: Lineage Tracing. Methods in Molecular Biology, vol. 2886, 139–151.'
mla: Cheung, Giselle T., et al. “Probing Cell-Type Specificity of Mutant Phenotype
at Transcriptomic Level Using Mosaic Analysis with Double Markers (MADM).” Lineage
Tracing, edited by Jorge Garcia-Marques and Tzumin Lee, vol. 2886, Springer
Nature, 2025, pp. 139–51, doi:10.1007/978-1-0716-4310-5_7.
short: G.T. Cheung, F. Pauler, S. Hippenmeyer, in:, J. Garcia-Marques, T. Lee (Eds.),
Lineage Tracing, Springer Nature, New York, NY, 2025, pp. 139–151.
corr_author: '1'
date_created: 2025-01-07T08:36:47Z
date_published: 2025-01-03T00:00:00Z
date_updated: 2025-04-14T07:43:46Z
day: '03'
department:
- _id: SiHi
doi: 10.1007/978-1-0716-4310-5_7
ec_funded: 1
editor:
- first_name: Jorge
full_name: Garcia-Marques, Jorge
last_name: Garcia-Marques
- first_name: Tzumin
full_name: Lee, Tzumin
last_name: Lee
external_id:
pmid:
- '39745639'
fulldoi: https://doi.org/10.1007/978-1-0716-4310-5_7
intvolume: ' 2886'
language:
- iso: eng
month: '01'
oa_version: None
page: 139-151
place: New York, NY
pmid: 1
project:
- _id: 260C2330-B435-11E9-9278-68D0E5697425
call_identifier: H2020
grant_number: '754411'
name: ISTplus - Postdoctoral Fellowships
publication: Lineage Tracing
publication_identifier:
eisbn:
- '9781071643105'
eissn:
- 1940-6029
isbn:
- '9781071643099'
issn:
- 1064-3745
publication_status: published
publisher: Springer Nature
quality_controlled: '1'
scopus_import: '1'
series_title: MIMB
status: public
title: Probing Cell-Type Specificity of Mutant Phenotype at Transcriptomic Level Using
Mosaic Analysis with Double Markers (MADM)
type: book_chapter
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 2886
year: '2025'
...
---
_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: Yes (in subscription journal)
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. STAR Protocols.
2024;5(1). doi:10.1016/j.xpro.2023.102771
apa: Amberg, N., Cheung, G. T., & Hippenmeyer, S. (2024). Protocol for sorting
cells from mouse brains labeled with mosaic analysis with double markers by flow
cytometry. STAR Protocols. Elsevier. https://doi.org/10.1016/j.xpro.2023.102771
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.” STAR Protocols. Elsevier, 2024. https://doi.org/10.1016/j.xpro.2023.102771.
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,”
STAR Protocols, vol. 5, no. 1. Elsevier, 2024.
ista: Amberg N, Cheung GT, Hippenmeyer S. 2024. 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.” STAR Protocols,
vol. 5, no. 1, 102771, Elsevier, 2024, doi:10.1016/j.xpro.2023.102771.
short: N. Amberg, G.T. Cheung, S. Hippenmeyer, STAR Protocols 5 (2024).
corr_author: '1'
das_tickbox: '1'
dataavailabilitystatement: No custom code or dataset was generated in this protocol.
date_created: 2023-12-13T11:48:05Z
date_published: 2024-03-15T00:00:00Z
date_updated: 2026-10-21T08:40:15Z
day: '15'
ddc:
- '570'
department:
- _id: SiHi
doi: 10.1016/j.xpro.2023.102771
ec_funded: 1
external_id:
pmid:
- '38070137'
file:
- access_level: open_access
checksum: 3f0ee62e04bf5a44b45b035662826e95
content_type: application/pdf
creator: dernst
date_created: 2024-07-16T11:50:03Z
date_updated: 2024-07-16T11:50:03Z
file_id: '17260'
file_name: 2024_STARProtoc_Amberg.pdf
file_size: 8871807
relation: main_file
success: 1
file_date_updated: 2024-07-16T11:50:03Z
fulldoi: https://doi.org/10.1016/j.xpro.2023.102771
has_accepted_license: '1'
intvolume: ' 5'
issue: '1'
keyword:
- General Immunology and Microbiology
- General Biochemistry
- Genetics and Molecular Biology
- General Neuroscience
language:
- iso: eng
month: '03'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: 268F8446-B435-11E9-9278-68D0E5697425
call_identifier: FWF
grant_number: T01031
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: F7805
name: Stem Cell Modulation in Neural Development and Regeneration/ P05-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: published
publisher: Elsevier
quality_controlled: '1'
researchdata_availability: not applicable
scopus_import: '1'
status: public
supplementarymaterial: no
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: 317138e5-6ab7-11ef-aa6d-ffef3953e345
volume: 5
year: '2024'
...
---
APC_amount: 804 EUR
OA_place: publisher
OA_type: gold
_id: '17187'
abstract:
- lang: eng
text: "The generation of diverse cell types during development is fundamental to
brain\r\nfunctions. We outline a protocol to quantitatively assess the clonal
output of individual neural progenitors using mosaic analysis with double markers
(MADM) in\r\nmice. We first describe steps to acquire and reconstruct adult MADM
clones in\r\nthe superior colliculus. Then we detail analysis pipelines to determine
clonal\r\ncomposition and architecture. This protocol enables the buildup of quantitative\r\nframeworks
of lineage progression with precise spatial resolution in the brain.\r\nFor complete
details on the use and execution of this protocol, please refer to\r\nCheung et
al.1"
acknowledged_ssus:
- _id: Bio
- _id: PreCl
acknowledgement: We thank A. Heger for mouse breeding support. This work was supported
by the Scientific Service Units of IST Austria through resources provided by the
Imaging & Optics and Preclinical facilities. G.C. received funding from the European
Commission (IST plus postdoctoral fellowship); S.H. was funded by ISTA institutional
funds and the Austrian Science Fund Special Research Programmes (FWF SFB-F78 Neuro
Stem Modulation).
article_number: '103157'
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: Carmen
full_name: Streicher, Carmen
id: 36BCB99C-F248-11E8-B48F-1D18A9856A87
last_name: Streicher
- first_name: Simon
full_name: Hippenmeyer, Simon
id: 37B36620-F248-11E8-B48F-1D18A9856A87
last_name: Hippenmeyer
orcid: 0000-0003-2279-1061
citation:
ama: Cheung GT, Streicher C, Hippenmeyer S. Protocol for quantitative reconstruction
of cell lineage using mosaic analysis with double markers in mice. STAR Protocols.
2024;5(3). doi:10.1016/j.xpro.2024.103157
apa: Cheung, G. T., Streicher, C., & Hippenmeyer, S. (2024). Protocol for quantitative
reconstruction of cell lineage using mosaic analysis with double markers in mice.
STAR Protocols. Elsevier. https://doi.org/10.1016/j.xpro.2024.103157
chicago: Cheung, Giselle T, Carmen Streicher, and Simon Hippenmeyer. “Protocol for
Quantitative Reconstruction of Cell Lineage Using Mosaic Analysis with Double
Markers in Mice.” STAR Protocols. Elsevier, 2024. https://doi.org/10.1016/j.xpro.2024.103157.
ieee: G. T. Cheung, C. Streicher, and S. Hippenmeyer, “Protocol for quantitative
reconstruction of cell lineage using mosaic analysis with double markers in mice,”
STAR Protocols, vol. 5, no. 3. Elsevier, 2024.
ista: Cheung GT, Streicher C, Hippenmeyer S. 2024. Protocol for quantitative reconstruction
of cell lineage using mosaic analysis with double markers in mice. STAR Protocols.
5(3), 103157.
mla: Cheung, Giselle T., et al. “Protocol for Quantitative Reconstruction of Cell
Lineage Using Mosaic Analysis with Double Markers in Mice.” STAR Protocols,
vol. 5, no. 3, 103157, Elsevier, 2024, doi:10.1016/j.xpro.2024.103157.
short: G.T. Cheung, C. Streicher, S. Hippenmeyer, STAR Protocols 5 (2024).
corr_author: '1'
das_tickbox: '1'
dataavailabilitystatement: No dataset was generated while composing this protocol.
A FIJI macro script “3DSerialSectionAlignmentLineROI.ijm” and a Microsoft Excel
template used in this protocol are provided as supplementary materials.
date_created: 2024-06-30T22:01:04Z
date_published: 2024-09-20T00:00:00Z
date_updated: 2026-10-21T09:14:52Z
day: '20'
ddc:
- '570'
department:
- _id: SiHi
doi: 10.1016/j.xpro.2024.103157
ec_funded: 1
external_id:
pmid:
- '38935508'
file:
- access_level: open_access
checksum: d8a8cdba82a394e731aa699ace1ae433
content_type: application/pdf
creator: dernst
date_created: 2025-01-09T12:12:40Z
date_updated: 2025-01-09T12:12:40Z
file_id: '18809'
file_name: 2024_STARProtoc_Cheung.pdf
file_size: 5186071
relation: main_file
success: 1
file_date_updated: 2025-01-09T12:12:40Z
fulldoi: https://doi.org/10.1016/j.xpro.2024.103157
has_accepted_license: '1'
intvolume: ' 5'
issue: '3'
language:
- iso: eng
month: '09'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: 260C2330-B435-11E9-9278-68D0E5697425
call_identifier: H2020
grant_number: '754411'
name: ISTplus - Postdoctoral Fellowships
- _id: 059F6AB4-7A3F-11EA-A408-12923DDC885E
grant_number: F7805
name: Stem Cell Modulation in Neural Development and Regeneration/ P05-Molecular
Mechanisms of Neural Stem Cell Lineage Progression
publication: STAR Protocols
publication_identifier:
eissn:
- 2666-1667
publication_status: published
publisher: Elsevier
quality_controlled: '1'
researchdata_availability: yes (supplementary data)
scopus_import: '1'
status: public
supplementarymaterial: yes
title: Protocol for quantitative reconstruction of cell lineage using mosaic analysis
with double markers in mice
tmp:
image: /images/cc_by_nc_nd.png
legal_code_url: https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode
name: Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
(CC BY-NC-ND 4.0)
short: CC BY-NC-ND (4.0)
type: journal_article
user_id: 317138e5-6ab7-11ef-aa6d-ffef3953e345
volume: 5
year: '2024'
...
---
APC_amount: 804 EUR
OA_place: publisher
OA_type: gold
_id: '17232'
abstract:
- lang: eng
text: "The lineage relationship of clonally-related cells offers important insights
into the ontogeny and cytoarchitecture of the brain in health and disease. Here,
we provide a protocol to concurrently assess cell lineage relationship and cell-type
identity among clonally-related cells in situ. We first describe the preparation
and screening of acute brain slices containing clonally-related cells labeled
using mosaic analysis with double markers (MADM). We then outline steps to collect
RNA from individual cells for downstream applications and cell-type identification
using RNA sequencing.\r\nFor complete details on the use and execution of this
protocol, please refer to Cheung et al.\r\n1"
acknowledged_ssus:
- _id: Bio
- _id: M-Shop
- _id: PreCl
acknowledgement: We thank R. Beattie and T. Asenov for designing and producing components
of the multi-well slice recover chamber. We thank R. Shigemoto for providing equipment
access. We thank C. Streicher and A. Heger for mouse breeding support. This work
was supported by the Scientific Service Units of IST Austria through resources provided
by the Imaging & Optics, Miba Machine Shop, and Preclinical facilities. G.C. received
funding from the European Commission (IST plus postdoctoral fellowship) and S.H.
was funded by ISTA institutional funds and the Austrian Science Fund Special Research
Programmes (FWF SFB-F78 Neuro Stem Modulation).
article_number: '103168'
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: 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: Simon
full_name: Hippenmeyer, Simon
id: 37B36620-F248-11E8-B48F-1D18A9856A87
last_name: Hippenmeyer
orcid: 0000-0003-2279-1061
citation:
ama: Cheung GT, Pauler F, Koppensteiner P, Hippenmeyer S. Protocol for mapping cell
lineage and cell-type identity of clonally-related cells in situ using MADM-CloneSeq.
STAR Protocols. 2024;5(3). doi:10.1016/j.xpro.2024.103168
apa: Cheung, G. T., Pauler, F., Koppensteiner, P., & Hippenmeyer, S. (2024).
Protocol for mapping cell lineage and cell-type identity of clonally-related cells
in situ using MADM-CloneSeq. STAR Protocols. Elsevier. https://doi.org/10.1016/j.xpro.2024.103168
chicago: Cheung, Giselle T, Florian Pauler, Peter Koppensteiner, and Simon Hippenmeyer.
“Protocol for Mapping Cell Lineage and Cell-Type Identity of Clonally-Related
Cells in Situ Using MADM-CloneSeq.” STAR Protocols. Elsevier, 2024. https://doi.org/10.1016/j.xpro.2024.103168.
ieee: G. T. Cheung, F. Pauler, P. Koppensteiner, and S. Hippenmeyer, “Protocol for
mapping cell lineage and cell-type identity of clonally-related cells in situ
using MADM-CloneSeq,” STAR Protocols, vol. 5, no. 3. Elsevier, 2024.
ista: Cheung GT, Pauler F, Koppensteiner P, Hippenmeyer S. 2024. Protocol for mapping
cell lineage and cell-type identity of clonally-related cells in situ using MADM-CloneSeq.
STAR Protocols. 5(3), 103168.
mla: Cheung, Giselle T., et al. “Protocol for Mapping Cell Lineage and Cell-Type
Identity of Clonally-Related Cells in Situ Using MADM-CloneSeq.” STAR Protocols,
vol. 5, no. 3, 103168, Elsevier, 2024, doi:10.1016/j.xpro.2024.103168.
short: G.T. Cheung, F. Pauler, P. Koppensteiner, S. Hippenmeyer, STAR Protocols
5 (2024).
corr_author: '1'
das_tickbox: '1'
dataavailabilitystatement: No dataset or new code was generated while composing this
protocol.
date_created: 2024-07-14T22:01:10Z
date_published: 2024-09-20T00:00:00Z
date_updated: 2026-10-21T09:15:51Z
day: '20'
ddc:
- '570'
department:
- _id: SiHi
- _id: PreCl
doi: 10.1016/j.xpro.2024.103168
external_id:
pmid:
- '38968076'
file:
- access_level: open_access
checksum: 464f52ecc6ec92f509552823bb82bf79
content_type: application/pdf
creator: dernst
date_created: 2025-01-09T12:16:53Z
date_updated: 2025-01-09T12:16:53Z
file_id: '18810'
file_name: 2024_STARProtoc_Cheung2.pdf
file_size: 6445556
relation: main_file
success: 1
file_date_updated: 2025-01-09T12:16:53Z
fulldoi: https://doi.org/10.1016/j.xpro.2024.103168
has_accepted_license: '1'
intvolume: ' 5'
issue: '3'
language:
- iso: eng
month: '09'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: 059F6AB4-7A3F-11EA-A408-12923DDC885E
grant_number: F7805
name: Stem Cell Modulation in Neural Development and Regeneration/ P05-Molecular
Mechanisms of Neural Stem Cell Lineage Progression
publication: STAR Protocols
publication_identifier:
eissn:
- 2666-1667
publication_status: published
publisher: Elsevier
quality_controlled: '1'
researchdata_availability: not applicable
scopus_import: '1'
status: public
supplementarymaterial: no
title: Protocol for mapping cell lineage and cell-type identity of clonally-related
cells in situ using MADM-CloneSeq
tmp:
image: /images/cc_by_nc_nd.png
legal_code_url: https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode
name: Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
(CC BY-NC-ND 4.0)
short: CC BY-NC-ND (4.0)
type: journal_article
user_id: 317138e5-6ab7-11ef-aa6d-ffef3953e345
volume: 5
year: '2024'
...
---
_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: 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: 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
citation:
ama: Cheung GT, Pauler F, Koppensteiner P, et al. Multipotent progenitors instruct
ontogeny of the superior colliculus. Neuron. 2024;112(2):230-246.e11. doi:10.1016/j.neuron.2023.11.009
apa: Cheung, G. T., Pauler, F., Koppensteiner, P., Krausgruber, T., Streicher, C.,
Schrammel, M., … Hippenmeyer, S. (2024). Multipotent progenitors instruct ontogeny
of the superior colliculus. Neuron. Elsevier. https://doi.org/10.1016/j.neuron.2023.11.009
chicago: Cheung, Giselle T, Florian Pauler, Peter Koppensteiner, Thomas Krausgruber,
Carmen Streicher, Martin Schrammel, Natalie Y Özgen, et al. “Multipotent Progenitors
Instruct Ontogeny of the Superior Colliculus.” Neuron. Elsevier, 2024.
https://doi.org/10.1016/j.neuron.2023.11.009.
ieee: G. T. Cheung et al., “Multipotent progenitors instruct ontogeny of
the superior colliculus,” Neuron, vol. 112, no. 2. Elsevier, p. 230–246.e11,
2024.
ista: Cheung GT, Pauler F, Koppensteiner P, Krausgruber T, Streicher C, Schrammel
M, Özgen NY, Ivec A, Bock C, Shigemoto R, Hippenmeyer S. 2024. Multipotent progenitors
instruct ontogeny of the superior colliculus. Neuron. 112(2), 230–246.e11.
mla: Cheung, Giselle T., et al. “Multipotent Progenitors Instruct Ontogeny of the
Superior Colliculus.” Neuron, vol. 112, no. 2, Elsevier, 2024, p. 230–246.e11,
doi:10.1016/j.neuron.2023.11.009.
short: G.T. Cheung, F. Pauler, P. Koppensteiner, T. Krausgruber, C. Streicher, M.
Schrammel, N.Y. Özgen, A. Ivec, C. Bock, R. Shigemoto, S. Hippenmeyer, Neuron
112 (2024) 230–246.e11.
corr_author: '1'
das_tickbox: '1'
dataavailabilitystatement: All data generated and analyzed in this study are included
in the paper and/or supplementary materials. Raw sequencing data has been deposited
at Gene Expression Omnibus (GEO). All scripts that were used to prepare data and
figures for this manuscript are available via GitHub at https://github.com/fpauler.
Accession numbers and github DOI are listed in the key resources table. Files for
3D printing of acute slice recovery chamber (3D model 361319) are deposited to https://www.printables.com.
date_created: 2023-04-27T09:41:48Z
date_published: 2024-01-17T00:00:00Z
date_updated: 2026-10-21T09:51:58Z
day: '17'
ddc:
- '570'
department:
- _id: SiHi
- _id: RySh
doi: 10.1016/j.neuron.2023.11.009
external_id:
isi:
- '001163937900001'
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
fulldoi: https://doi.org/10.1016/j.neuron.2023.11.009
has_accepted_license: '1'
intvolume: ' 112'
isi: 1
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: F7805
name: Stem Cell Modulation in Neural Development and Regeneration/ P05-Molecular
Mechanisms of Neural Stem Cell Lineage Progression
publication: Neuron
publication_identifier:
issn:
- 0896-6273
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/
researchdata_availability: yes
scopus_import: '1'
status: public
supplementarymaterial: yes
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: 317138e5-6ab7-11ef-aa6d-ffef3953e345
volume: 112
year: '2024'
...
---
_id: '17425'
abstract:
- lang: eng
text: Mosaic Analysis with Double Markers (MADM) is a powerful genetic method typically
used for lineage tracing and to disentangle cell autonomous and tissue-wide roles
of candidate genes with single cell resolution. Given the relatively sparse labeling,
depending on which of the 19 MADM chromosomes one chooses, the MADM approach represents
the perfect opportunity for cell morphology analysis. Various MADM studies include
reports of morphological anomalies and phenotypes in the central nervous system
(CNS). MADM for any candidate gene can easily incorporate morphological analysis
within the experimental workflow. Here, we describe the methods of morphological
cell analysis which we developed in the course of diverse recent MADM studies.
This chapter will specifically focus on methods to quantify aspects of the morphology
of neurons and astrocytes within the CNS, but these methods can broadly be applied
to any MADM-labeled cells throughout the entire organism. We will cover two analyses—soma
volume and dendrite characterization—of physical characteristics of pyramidal
neurons in the somatosensory cortex, and two analyses—volume and Sholl analysis—of
astrocyte morphology.
acknowledged_ssus:
- _id: Bio
acknowledgement: We thank all Hippenmeyer lab members for support and discussions.
This work was supported by the Scientific Service Units (SSU) at ISTA through resources
provided by the Imaging & Optics Facility (IOF). O.A.M was a recipient of a DOC
Fellowship (26253) of the Austrian Academy of Sciences. This work was supported
by ISTA institutional funds, and The Austrian Science Fund Special Research Programmes
(FWF SFB F78 Neuro Stem Modulation) to S.H.
alternative_title:
- Methods in Molecular Biology
article_processing_charge: No
author:
- first_name: Osvaldo
full_name: Miranda, Osvaldo
id: 862A3C56-A8BF-11E9-B4FA-D9E3E5697425
last_name: Miranda
orcid: 0000-0001-6618-6889
- 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: 'Miranda O, Cheung GT, Hippenmeyer S. Morphological Analysis of Neurons and
Glia Using Mosaic Analysis with Double Markers. In: Toyooka K, ed. Neuronal
Morphogenesis. Vol 2831. 1st ed. MIMB. New York, NY: Springer Nature; 2024:283-299.
doi:10.1007/978-1-0716-3969-6_19'
apa: 'Miranda, O., Cheung, G. T., & Hippenmeyer, S. (2024). Morphological Analysis
of Neurons and Glia Using Mosaic Analysis with Double Markers. In K. Toyooka (Ed.),
Neuronal Morphogenesis (1st ed., Vol. 2831, pp. 283–299). New York, NY:
Springer Nature. https://doi.org/10.1007/978-1-0716-3969-6_19'
chicago: 'Miranda, Osvaldo, Giselle T Cheung, and Simon Hippenmeyer. “Morphological
Analysis of Neurons and Glia Using Mosaic Analysis with Double Markers.” In Neuronal
Morphogenesis, edited by Kazuhito Toyooka, 1st ed., 2831:283–99. MIMB. New
York, NY: Springer Nature, 2024. https://doi.org/10.1007/978-1-0716-3969-6_19.'
ieee: 'O. Miranda, G. T. Cheung, and S. Hippenmeyer, “Morphological Analysis of
Neurons and Glia Using Mosaic Analysis with Double Markers,” in Neuronal Morphogenesis,
1st ed., vol. 2831, K. Toyooka, Ed. New York, NY: Springer Nature, 2024, pp. 283–299.'
ista: 'Miranda O, Cheung GT, Hippenmeyer S. 2024.Morphological Analysis of Neurons
and Glia Using Mosaic Analysis with Double Markers. In: Neuronal Morphogenesis.
Methods in Molecular Biology, vol. 2831, 283–299.'
mla: Miranda, Osvaldo, et al. “Morphological Analysis of Neurons and Glia Using
Mosaic Analysis with Double Markers.” Neuronal Morphogenesis, edited by
Kazuhito Toyooka, 1st ed., vol. 2831, Springer Nature, 2024, pp. 283–99, doi:10.1007/978-1-0716-3969-6_19.
short: O. Miranda, G.T. Cheung, S. Hippenmeyer, in:, K. Toyooka (Ed.), Neuronal
Morphogenesis, 1st ed., Springer Nature, New York, NY, 2024, pp. 283–299.
corr_author: '1'
das_tickbox: '0'
date_created: 2024-08-13T12:16:41Z
date_published: 2024-08-13T00:00:00Z
date_updated: 2026-10-22T22:31:24Z
day: '13'
department:
- _id: GradSch
- _id: SiHi
doi: 10.1007/978-1-0716-3969-6_19
edition: '1'
editor:
- first_name: Kazuhito
full_name: Toyooka, Kazuhito
last_name: Toyooka
external_id:
pmid:
- '39134857'
fulldoi: https://doi.org/10.1007/978-1-0716-3969-6_19
intvolume: ' 2831'
language:
- iso: eng
month: '08'
oa_version: None
page: 283-299
place: New York, NY
pmid: 1
project:
- _id: 34c9fbcb-11ca-11ed-8bc3-98fa5658610d
grant_number: '26253'
name: Molecular Mechanisms Regulating Cortical Neural Stem Cell Lineage Progression
and Astrocyte Development
- _id: 059F6AB4-7A3F-11EA-A408-12923DDC885E
grant_number: F7805
name: Stem Cell Modulation in Neural Development and Regeneration/ P05-Molecular
Mechanisms of Neural Stem Cell Lineage Progression
publication: Neuronal Morphogenesis
publication_identifier:
eisbn:
- '9781071639696'
eissn:
- 1940-6029
isbn:
- '9781071639689'
issn:
- 1064-3745
publication_status: published
publisher: Springer Nature
quality_controlled: '1'
related_material:
record:
- id: '20212'
relation: dissertation_contains
status: public
researchdata_availability: no
scopus_import: '1'
series_title: MIMB
status: public
supplementarymaterial: yes
title: Morphological Analysis of Neurons and Glia Using Mosaic Analysis with Double
Markers
type: book_chapter
user_id: 317138e5-6ab7-11ef-aa6d-ffef3953e345
volume: 2831
year: '2024'
...
---
_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. Cells. 2023;12(8). doi:10.3390/cells12081133
apa: Cheung, G. T., Chever, O., Rollenhagen, A., Quenech’du, N., Ezan, P., Lübke,
J. H. R., & Rouach, N. (2023). Astroglial connexin 43 regulates synaptic vesicle
release at hippocampal synapses. Cells. MDPI. https://doi.org/10.3390/cells12081133
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.” Cells. MDPI,
2023. https://doi.org/10.3390/cells12081133.
ieee: G. T. Cheung et al., “Astroglial connexin 43 regulates synaptic vesicle
release at hippocampal synapses,” Cells, 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.” Cells, vol. 12, no. 8, 1133, MDPI, 2023,
doi:10.3390/cells12081133.
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
fulldoi: https://doi.org/10.3390/cells12081133
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
orcid: 0000-0001-8457-2572
- 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. Nature Communications.
2022;13. doi:10.1038/s41467-022-28331-7
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. Nature Communications. Springer Nature. https://doi.org/10.1038/s41467-022-28331-7
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.” Nature Communications.
Springer Nature, 2022. https://doi.org/10.1038/s41467-022-28331-7.
ieee: G. T. Cheung et al., “Physiological synaptic activity and recognition
memory require astroglial glutamine,” Nature Communications, 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.” Nature Communications, vol. 13, 753,
Springer Nature, 2022, doi:10.1038/s41467-022-28331-7.
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: 2026-04-02T12:22:38Z
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
file_id: '10777'
file_name: 2022_NatureCommunications_Cheung.pdf
file_size: 7910519
relation: main_file
success: 1
file_date_updated: 2022-02-21T07:51:33Z
fulldoi: https://doi.org/10.1038/s41467-022-28331-7
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:
- 2041-1723
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: ba8df636-2132-11f1-aed0-ed93e2281fdd
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).
Journal of Visual Experiments. 2020;(159). doi:10.3791/61147
apa: Beattie, R. J., Streicher, C., Amberg, N., Cheung, G. T., Contreras, X., Hansen,
A. H., & Hippenmeyer, S. (2020). Lineage tracing and clonal analysis in developing
cerebral cortex using mosaic analysis with double markers (MADM). Journal of
Visual Experiments. MyJove Corporation. https://doi.org/10.3791/61147
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).”
Journal of Visual Experiments. MyJove Corporation, 2020. https://doi.org/10.3791/61147.
ieee: R. J. Beattie et al., “Lineage tracing and clonal analysis in developing
cerebral cortex using mosaic analysis with double markers (MADM),” Journal
of Visual Experiments, 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).” Journal
of Visual Experiments, no. 159, e61147, MyJove Corporation, 2020, doi:10.3791/61147.
short: R.J. Beattie, C. Streicher, N. Amberg, G.T. Cheung, X. Contreras, A.H. Hansen,
S. Hippenmeyer, Journal of Visual Experiments (2020).
corr_author: '1'
date_created: 2020-05-11T08:31:20Z
date_published: 2020-05-08T00:00:00Z
date_updated: 2026-10-22T22:30:21Z
day: '08'
ddc:
- '570'
department:
- _id: SiHi
doi: 10.3791/61147
ec_funded: 1
external_id:
isi:
- '000546406600043'
pmid:
- '32449730'
file:
- access_level: open_access
checksum: 3154ea7f90b9fb45e084cd1c2770597d
content_type: application/pdf
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
fulldoi: https://doi.org/10.3791/61147
has_accepted_license: '1'
isi: 1
issue: '159'
language:
- iso: eng
month: '05'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: 264E56E2-B435-11E9-9278-68D0E5697425
call_identifier: FWF
grant_number: M02416
name: Molecular Mechanisms Regulating Gliogenesis in the Neocortex
- _id: 268F8446-B435-11E9-9278-68D0E5697425
call_identifier: FWF
grant_number: T01031
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:
record:
- id: '7902'
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.
Journal of Neurochemistry. 2019;151(5):570-583. doi:10.1111/jnc.14862
apa: Cheung, G. T., & Cousin, M. A. (2019). Synaptic vesicle generation from
activity‐dependent bulk endosomes requires a dephosphorylation‐dependent dynamin–syndapin
interaction. Journal of Neurochemistry. Wiley. https://doi.org/10.1111/jnc.14862
chicago: Cheung, Giselle T, and Michael A. Cousin. “Synaptic Vesicle Generation
from Activity‐dependent Bulk Endosomes Requires a Dephosphorylation‐dependent
Dynamin–Syndapin Interaction.” Journal of Neurochemistry. Wiley, 2019.
https://doi.org/10.1111/jnc.14862.
ieee: G. T. Cheung and M. A. Cousin, “Synaptic vesicle generation from activity‐dependent
bulk endosomes requires a dephosphorylation‐dependent dynamin–syndapin interaction,”
Journal of Neurochemistry, 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.” Journal of Neurochemistry, vol. 151, no. 5, Wiley, 2019,
pp. 570–83, doi:10.1111/jnc.14862.
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:
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title: Synaptic vesicle generation from activity‐dependent bulk endosomes requires
a dephosphorylation‐dependent dynamin–syndapin interaction
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