Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Accepted manuscripts
    • Issue in progress
    • Latest complete issue
    • Issue archive
    • Archive by article type
    • Special issues
    • Subject collections
    • Sign up for alerts
  • About us
    • About Development
    • About the Node
    • Editors and Board
    • Editor biographies
    • Travelling Fellowships
    • Grants and funding
    • Journal Meetings
    • Workshops
    • The Company of Biologists
    • Journal news
  • For authors
    • Submit a manuscript
    • Aims and scope
    • Presubmission enquiries
    • Article types
    • Manuscript preparation
    • Cover suggestions
    • Editorial process
    • Promoting your paper
    • Open Access
    • Biology Open transfer
  • Journal info
    • Journal policies
    • Rights and permissions
    • Media policies
    • Reviewer guide
    • Sign up for alerts
  • Contacts
    • Contacts
    • Subscriptions
    • Advertising
    • Feedback
    • Institutional usage stats (logged-in users only)
  • COB
    • About The Company of Biologists
    • Development
    • Journal of Cell Science
    • Journal of Experimental Biology
    • Disease Models & Mechanisms
    • Biology Open

User menu

  • Log in

Search

  • Advanced search
Development
  • COB
    • About The Company of Biologists
    • Development
    • Journal of Cell Science
    • Journal of Experimental Biology
    • Disease Models & Mechanisms
    • Biology Open

supporting biologistsinspiring biology

Development

  • Log in
Advanced search

RSS  Twitter  Facebook  YouTube 

  • Home
  • Articles
    • Accepted manuscripts
    • Issue in progress
    • Latest complete issue
    • Issue archive
    • Archive by article type
    • Special issues
    • Subject collections
    • Sign up for alerts
  • About us
    • About Development
    • About the Node
    • Editors and Board
    • Editor biographies
    • Travelling Fellowships
    • Grants and funding
    • Journal Meetings
    • Workshops
    • The Company of Biologists
    • Journal news
  • For authors
    • Submit a manuscript
    • Aims and scope
    • Presubmission enquiries
    • Article types
    • Manuscript preparation
    • Cover suggestions
    • Editorial process
    • Promoting your paper
    • Open Access
    • Biology Open transfer
  • Journal info
    • Journal policies
    • Rights and permissions
    • Media policies
    • Reviewer guide
    • Sign up for alerts
  • Contacts
    • Contacts
    • Subscriptions
    • Advertising
    • Feedback
    • Institutional usage stats (logged-in users only)
RESEARCH ARTICLE
Fat1 interacts with Fat4 to regulate neural tube closure, neural progenitor proliferation and apical constriction during mouse brain development
Caroline Badouel, Mark A. Zander, Nicole Liscio, Mazdak Bagherie-Lachidan, Richelle Sopko, Etienne Coyaud, Brian Raught, Freda D. Miller, Helen McNeill
Development 2015 142: 2781-2791; doi: 10.1242/dev.123539
Caroline Badouel
1Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, M5G 1X5, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Mark A. Zander
2Neuroscience and Mental Health Program, Hospital for Sick Children, University of Toronto, Toronto, Ontario, Canada
3Institute of Medical Sciences, University of Toronto, Toronto, Ontario, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Nicole Liscio
1Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, M5G 1X5, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Mazdak Bagherie-Lachidan
1Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, M5G 1X5, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Richelle Sopko
4Department of Genetics, Harvard Medical School, Boston, MA 02115, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Etienne Coyaud
5Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, M5G 2M9, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Brian Raught
5Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, M5G 2M9, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Freda D. Miller
2Neuroscience and Mental Health Program, Hospital for Sick Children, University of Toronto, Toronto, Ontario, Canada
3Institute of Medical Sciences, University of Toronto, Toronto, Ontario, Canada
6Department of Molecular Genetics, University of Toronto, Toronto, Ontario, M5S 1A8, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Helen McNeill
1Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, M5G 1X5, Canada
6Department of Molecular Genetics, University of Toronto, Toronto, Ontario, M5S 1A8, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: mcneill@lunenfeld.ca
  • Article
  • Figures & tables
  • Supp info
  • Info & metrics
  • PDF + SI
  • PDF
Loading

Article Figures & Tables

Figures

  • Fig. 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 1.

    Loss of Fat1 leads to cranial neural tube defects. (A) Schematic of mouse cortex development. Radial precursors are bipolar cells that can divide symmetrically to self-renew, or asymmetrically, producing either an intermediate neuronal progenitor, or a neuron that can migrate along radial precursor basal processes to form the new neuronal layers of the cortex. (B) Fat1 expression in a coronal section from E13.5 brain revealed by in situ hybridization. The left panel is a magnification of the cortex. Note that Fat1 is strongly expressed in the VZ of the cortex containing radial precursors. (C) Coronal section through E14.5 Fat1+/− brain stained for β-galactosidase to assess Fat1-lacZ expression. (D) Coronal section through E14.5 brain stained for β-galactosidase (red) and the cortical precursor marker Pax6 (green) and with Hoechst (blue). (E) Dorso-anterior view of an E11.5 Fat1−/− embryo and a control sibling. Note that anterior neural folds are open in Fat1−/− (arrow). (F) E13.5 Fat1−/− embryo showing exencephaly and control sibling. VZ, ventricular zone; SVZ, subventricular zone; IZ, intermediate zone; CP, cortical plate; V, ventricle. Scale bars: 100 µm in B; 500 µm in C; 40 µm in D.

  • Fig. 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 2.

    Fat1 exencephalic cortex exhibits elongated ventricles and increased proliferation. (A) Coronal section through E14.5 Fat1−/− exencephalic brain and control sibling stained for the dorsal cortical precursor marker Pax6 (red, top), the pan-neuronal marker βIII-tubulin (green), the intermediate neuronal progenitor marker Tbr2 (red, bottom) and with Hoechst (blue). Note that the third ventricle (yellow dotted line) is open in Fat1−/− exencephalic brains in the thalamic region (diencephalon) and is expanded dorsally above and over the cortex (telencephalon); the lateral ventricles of the cortex are delineated with white dotted lines. (B,C) Dorsal ventricular length (B) and Pax6+ area (C) measured from sections as in A. ***P<0.001; n=3 Fat1−/− exencephalic mutants and n=4 controls. (D) Coronal sections through E14.5 Fat1−/− exencephalic cortex and a control sibling stained for the mitosis marker phospho-histone H3 (pHH3, red) and with Hoechst (blue). V, ventricle (delineated with white dotted line). (E) Percentage of pHH3+ cells in the cortex, calculated per unit ventricle length, using images similar to D. *P<0.05; n=3 embryos each. Data are represented as mean±s.e.m. Scale bars: 500 µm in A; 20 µm in D.

  • Fig. 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 3.

    Fat1 knockdown promotes radial glial precursor proliferation. (A) Schematic of the in utero electroporation technique. (B-R) Murine cortices were electroporated at E13.5 with a nuclear EGFP expression plasmid and Fat1 shRNA (shFat1) or a scrambled shRNA (Ctrl) plasmid, and analyzed 3 days later at E16/17. (B) Fluorescence confocal micrographs of E16.5 cortex stained for EGFP (green). (C) Quantification of sections as in B for the percentage localization of EGFP+ cells (n=3 embryos each). (D) Confocal fluorescence micrographs of VZ/SVZ/IZ from coronal sections through E16.5 cortex immunostained for Pax6 (red) and EGFP (green); bottom panels show merge. (E,F) Quantification of sections as in D for the proportion of Pax6+, EGFP+ cells per section (E; n=3 embryos each) and relative localization in the VZ, SVZ or IZ (F; n=3 embryos each). (G) Confocal fluorescence micrographs of VZ/SVZ/IZ from coronal E16.5 cortical sections immunostained for Ki67 (red) and EGFP (green); bottom panels show merge. (H) Quantification of sections as in G for proportion of Ki67+, EGFP+ cells per section (n=3 embryos each). (I,J) Quantification of sections as in supplementary material Fig. S3B for the proportion of Pax6+, Ki67+, EGFP+ triple-labeled cells per section (I; n=3 embryos each) and relative localization in the VZ, SVZ or IZ (J; n=3 embryos each). (K) Fluorescence confocal micrographs of VZ/SVZ/IZ from coronal E16.5 cortical sections stained for Tbr2 (red) and EGFP (green); bottom panels show merge. (L) Quantification of sections as in K for the proportion of Tbr2+, EGFP+ cells per section (n=3 embryos each). (M) Quantification of sections as in supplementary material Fig. S3C for the proportion of Pax6+, Tbr2+, EGFP+ triple-labeled cells per section (n=3 embryos each). (N) Fluorescence confocal micrographs of CP from coronal E16.5 cortical sections immunostained for Satb2 (red) and EGFP (green); lower panels show merge. (O,P) Quantification of sections as in N and supplementary material Fig. S3D for the proportion of Satb2+, EGFP+ cells per section (O; n=3 embryos each) and relative localization in the VZ/SVZ/IZ versus CP (P; n=3 embryos each). (Q,R) Quantification of sections as in supplementary material Fig. S3E for the proportion of βIII-tubulin+, EGFP+ cells per section (Q; n=3 embryos each) and the relative localization in the VZ/SVZ/IZ versus CP (R; n=3 embryos each). Arrowheads indicate EGFP+ cells negative for the respective marker and arrows indicate double-labeled cells. *P<0.05, **P<0.01, ***P<0.001; ns, not significant; error bars denote s.e.m. Scale bars: 20 μm.

  • Fig. 4.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 4.

    Fat1-Fat4 mutant cortices display enhanced radial precursor proliferation and delayed cell cycle exit. E13.5 Fat1-Fat4 mutant embryos and control siblings were labeled with BrdU and harvested after 24 h. (A,C,E) Coronal sections through E14.5 dorsal cortex stained for BrdU (red) and Ki67 (A), Sox2 (C) and Tbr2 (E) (green); right panel shows merge with Hoechst (blue). Arrowheads point to examples of double-labeled cells (BrdU+ and Ki67+, Sox2+ or Tbr2+) and arrows to single-labeled BrdU+ cells. Scale bars: 50 μm. (B,D,F) Quantification of the percentage of Ki67–, BrdU+ (B), Sox2+, BrdU+ (D) and Tbr2+, BrdU+ (F) over total BrdU+ cells shows reduced cell cycle exit in Fat1-Fat4 mutants. *P<0.05, ***P<0.001; n=3 embryos each; data are mean±s.e.m.

  • Fig. 5.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 5.

    Fat1 and Fat4 have additive effects on radial glial precursor proliferation. (A-J) Murine cortices were electroporated with a nuclear EGFP expression plasmid and a control shRNA (Ctrl) or combined Fat1 shRNA and Fat4 shRNA at E13.5, and analyzed 3 days later at E16/17. (A) Confocal fluorescence micrographs of E16.5 coronal cortical sections stained for EGFP (green). (B) Quantification of sections as in A for the relative localization of EGFP+ cells (n=3 embryos each). (C) Confocal fluorescence micrographs of the VZ/SVZ/IZ of coronal cortical sections immunostained for EGFP (green) and Pax6 (red); right panels show merge. (D,E) Quantification of sections as in C for the proportion of Pax6+, EGFP+ cells per section (D; n=3 embryos each) and the relative localization in the VZ, SVZ or IZ (E; n=3 embryos each). (F) Confocal fluorescence micrographs of the VZ/SVZ/IZ of sections stained for EGFP (green) and Ki67 (red); right panels show merge. (G) Quantification of sections as in F for the proportion of EGFP+ cells that are Ki67+ (n=3 embryos each). (H) Confocal micrographs of the CP stained for EGFP (green) and Satb2 (red); right panels show merge. (I,J) Quantification of sections as in H for the proportion of EGFP+ cells that are Satb2+ (I; n=3 embryos each) and relative localization in VZ/SVZ/IZ versus CP (J; n=3 embryos each). Dotted lines demarcate cortical regions. Arrowheads indicate EGFP+ cells negative for the respective marker and arrows indicate double-labeled cells. *P<0.01, **P<0.01, ***P<0.001; error bars denote s.e.m. Scale bars: 20 μm.

  • Fig. 6.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 6.

    Fat1-Fat4 mutant radial glial precursors have apical constriction defects. (A) ZO-1 staining of thick coronal sections (10 µm). Beneath is a reconstruction of the apical surface area from z-stack confocal images. (B) Confocal fluorescence micrographs of the ventricular domain of cortical sections from E14.5 Fat1-Fat4 mutants and control sibling. Rhodamine-phalloidin was used to visualize F-actin. (C) Quantification of F-actin intensity from images as in B, showing a decrease in F-actin accumulation in Fat1-Fat4 mutants. **P<0.01; n=4 controls and n=3 Fat1-Fat4 mutants, at least 100 junctions were measured for each. (D) Reconstruction of radial precursor apical surface in Fat1−/−; Fat4−/− E14.5 cortex and control sibling stained for ZO-1 (green), Par3 (red) and Hoechst (blue). (E) Quantification of apical surface area. *P<0.05; n=7 controls and n=4 Fat1-Fat4 mutants, at least 100 cells were measured for each on at least two different stacks images. Scale bars: 10 µm in A,D; 20 µm in B. Data are mean±s.e.m.

  • Fig. 7.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 7.

    Fat1 and Fat4 form heterodimers and have different binding partners. (A) HEK293 cells stably expressing Fat1-ICD-Flag and Fat4-ICD-Flag constructs, or a control (GFP-Flag), were subjected to Flag immunoprecipitation followed by western blot with antibodies against Mena and Flag, as indicated. (B) HEK293 cells were co-transfected with an HA-tagged Fat1 construct missing most of its extracellular domain (Fat1-ΔECD-HA), together with Flag-tagged Fat4 constructs (Fat4-ΔECD-Flag or Fat4-ICD-Flag) or BMPRII as control. Flag immunoprecipitates were subject to western blot with antibodies against HA and Flag, as indicated.

Previous ArticleNext Article
Back to top
Previous ArticleNext Article

This Issue

Keywords

  • Fat cadherins
  • Mammalian cortex development
  • Apical constriction
  • Brain
  • Neural tube defects
  • Radial glial precursor

 Download PDF

Email

Thank you for your interest in spreading the word on Development.

NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.

Enter multiple addresses on separate lines or separate them with commas.
Fat1 interacts with Fat4 to regulate neural tube closure, neural progenitor proliferation and apical constriction during mouse brain development
(Your Name) has sent you a message from Development
(Your Name) thought you would like to see the Development web site.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
RESEARCH ARTICLE
Fat1 interacts with Fat4 to regulate neural tube closure, neural progenitor proliferation and apical constriction during mouse brain development
Caroline Badouel, Mark A. Zander, Nicole Liscio, Mazdak Bagherie-Lachidan, Richelle Sopko, Etienne Coyaud, Brian Raught, Freda D. Miller, Helen McNeill
Development 2015 142: 2781-2791; doi: 10.1242/dev.123539
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
RESEARCH ARTICLE
Fat1 interacts with Fat4 to regulate neural tube closure, neural progenitor proliferation and apical constriction during mouse brain development
Caroline Badouel, Mark A. Zander, Nicole Liscio, Mazdak Bagherie-Lachidan, Richelle Sopko, Etienne Coyaud, Brian Raught, Freda D. Miller, Helen McNeill
Development 2015 142: 2781-2791; doi: 10.1242/dev.123539

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Alerts

Please log in to add an alert for this article.

Sign in to email alerts with your email address

Article navigation

  • Top
  • Article
    • Abstract
    • INTRODUCTION
    • RESULTS
    • DISCUSSION
    • MATERIALS AND METHODS
    • Acknowledgements
    • Footnotes
    • References
  • Figures & tables
  • Supp info
  • Info & metrics
  • PDF + SI
  • PDF

Related articles

Cited by...

More in this TOC section

  • Precision of tissue patterning is controlled by dynamical properties of gene regulatory networks
  • Roles of developmentally regulated KIF2A alternative isoforms in cortical neuron migration and differentiation
  • Vasohibin 1 selectively regulates secondary sprouting and lymphangiogenesis in the zebrafish trunk
Show more RESEARCH ARTICLE

Similar articles

Other journals from The Company of Biologists

Journal of Cell Science

Journal of Experimental Biology

Disease Models & Mechanisms

Biology Open

Advertisement

An interview with Swathi Arur

Swathi Arur joined the team at Development as an Academic Editor in 2020. Her lab uses multidisciplinary approaches to understand female germline development and fertility. We met with her over Zoom to hear more about her life, her career and her love for C. elegans.


Jim Wells and Hanna Mikkola join our team of Editors

We are pleased to welcome James (Jim) Wells and Hanna Mikkola to our team of Editors. Jim joins us a new Academic Editor, taking over from Gordan Keller, and Hanna joins our team of Associate Editors. Find out more about their research interests and areas of expertise.


New funding scheme supports sustainable events

As part of our Sustainable Conferencing Initiative, we are pleased to announce funding for organisers that seek to reduce the environmental footprint of their event. The next deadline to apply for a Scientific Meeting grant is 26 March 2021.


Read & Publish participation continues to grow

“I’d heard of Read & Publish deals and knew that many universities, including mine, had signed up to them but I had not previously understood the benefits that these deals bring to authors who work at those universities.”

Professor Sally Lowell (University of Edinburgh) shares her experience of publishing Open Access as part of our growing Read & Publish initiative. We now have over 150 institutions in 15 countries and four library consortia taking part – find out more and view our full list of participating institutions.


Upcoming special issues

Imaging Development, Stem Cells and Regeneration
Submission deadline: 30 March 2021
Publication: mid-2021

The Immune System in Development and Regeneration
Guest editors: Florent Ginhoux and Paul Martin
Submission deadline: 1 September 2021
Publication: Spring 2022

Both special issues welcome Review articles as well as Research articles, and will be widely promoted online and at key global conferences.


Development presents...

Our successful webinar series continues into 2021, with early-career researchers presenting their papers and a chance to virtually network with the developmental biology community afterwards. Here, Brandon Carpenter talks about how inherited histone methylation defines the germline versus soma decision in C. elegans. 

Sign up to join our next session:

10 March
Time: TBC
Chaired by: Thomas Lecuit

Articles

  • Accepted manuscripts
  • Issue in progress
  • Latest complete issue
  • Issue archive
  • Archive by article type
  • Special issues
  • Subject collections
  • Sign up for alerts

About us

  • About Development
  • About the Node
  • Editors and board
  • Editor biographies
  • Travelling Fellowships
  • Grants and funding
  • Journal Meetings
  • Workshops
  • The Company of Biologists

For authors

  • Submit a manuscript
  • Aims and scope
  • Presubmission enquiries
  • Article types
  • Manuscript preparation
  • Cover suggestions
  • Editorial process
  • Promoting your paper
  • Open Access
  • Biology Open transfer

Journal info

  • Journal policies
  • Rights and permissions
  • Media policies
  • Reviewer guide
  • Sign up for alerts

Contact

  • Contact Development
  • Subscriptions
  • Advertising
  • Feedback
  • Institutional usage stats (logged-in users only)

 Twitter   YouTube   LinkedIn

© 2021   The Company of Biologists Ltd   Registered Charity 277992