---
_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
  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
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:
  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'
...