Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Accepted manuscripts
    • Issue in progress
    • Latest complete issue
    • Issue archive
    • Archive by article type
    • Subject collections
    • Interviews
    • Sign up for alerts
  • About us
    • About DMM
    • 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
    • Outstanding paper prize
    • Biology Open transfer
  • Journal info
    • Journal policies
    • Rights and permissions
    • Media policies
    • Reviewer guide
    • Sign up for alerts
  • Contact
    • Contact DMM
    • Advertising
    • Feedback
  • 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
Disease Models & Mechanisms
  • COB
    • About The Company of Biologists
    • Development
    • Journal of Cell Science
    • Journal of Experimental Biology
    • Disease Models & Mechanisms
    • Biology Open

supporting biologistsinspiring biology

Disease Models & Mechanisms

Advanced search

RSS   Twitter   Facebook   YouTube

  • Home
  • Articles
    • Accepted manuscripts
    • Issue in progress
    • Latest complete issue
    • Issue archive
    • Archive by article type
    • Subject collections
    • Interviews
    • Sign up for alerts
  • About us
    • About DMM
    • 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
    • Outstanding paper prize
    • Biology Open transfer
  • Journal info
    • Journal policies
    • Rights and permissions
    • Media policies
    • Reviewer guide
    • Sign up for alerts
  • Contact
    • Contact DMM
    • Advertising
    • Feedback
Research Article
Regulation of PDGFC signalling and extracellular matrix composition by FREM1 in mice
Fenny Wiradjaja, Denny L. Cottle, Lynelle Jones, Ian Smyth
Disease Models & Mechanisms 2013 6: 1426-1433; doi: 10.1242/dmm.013748
Fenny Wiradjaja
1Department of Biochemistry and Molecular Biology, Monash University, Wellington Road, Clayton, Victoria 3800, Australia
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Denny L. Cottle
1Department of Biochemistry and Molecular Biology, Monash University, Wellington Road, Clayton, Victoria 3800, Australia
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Lynelle Jones
1Department of Biochemistry and Molecular Biology, Monash University, Wellington Road, Clayton, Victoria 3800, Australia
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Ian Smyth
1Department of Biochemistry and Molecular Biology, Monash University, Wellington Road, Clayton, Victoria 3800, Australia
2Department of Anatomy and Developmental Biology, Monash University, Wellington Road, Clayton, Victoria 3800, Australia
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: ian.smyth@monash.edu
  • Article
  • Figures & tables
  • Info & metrics
  • PDF
Loading

Article Figures & Tables

Figures

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

    Interaction between FREM1 and PDGFC. (A) Structural representations of the full-length FREM1, FLAG-tagged construct and truncated constructs: NV alone, NV-CSPG, CSPG and CalXβ-C-lectin domains are shown with the tags indicated. The MYC tag shown refers to a 3×MYC tag + Igκ secretion signal. (ab), location of immunising sequence for anti-FREM1 antibodies upstream of mutation. (B) E13.5 WT and bat embryo head skin sections stained for FREM1 (green), PDGFC (red) and nuclear dye DAPI (blue). (C) NIH3T3 fibroblasts expressing FREM1-FLAG and PDGFC-V5 and immunostained as indicated. (D) Coimmunoprecipitation of FREM1-FLAG and PDGFC-V5 in transfected HEK293 cells. (E,F) Co-immunoprecipitation of PDGFC-V5 with MYC-tagged FREM1 subdomains. (G) Co-immunoprecipitation of endogenous FREM1 and PDGFC from embryo extracts at E12.5. A rabbit pre-immune serum was included as a control. * IgG heavy chain; IP, immunoprecipitation antibody; WB, western blotting antibody.

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

    FREM1 regulates activation of AKT and MAPK upon PDGFC stimulation. (A) Representative western blotting of phosphorylation of AKT and MAPK ERK1/2 in MEFs from WT and bat mouse embryos stimulated with PDGFCC for the indicated time periods. (B) Relative quantification of AKT phosphorylation levels. WT cells 10 minutes after stimulation were assigned a value of 1 and all other samples are standardised against this value. Graph represents average of up to nine WT and 16 bat samples, performed across four independent experiments from at least three different cell lines for each genotype. Black bars: WT; white bars, bat mutant. (C) FREM1 mutation in bat mutants reduces phosphorylation of PDGFRα in response to the addition of exogenous PDGFCC. IP, immunoprecipitation antibody; WB, western blotting antibody. (D) E13.5 WT embryo head skin sections stained for PDGFC (green), PDGFRα (red) and nuclear dye DAPI (blue). Error bars represent standard error of the mean (s.e.m.); *P<0.05, **P<0.01, ***P<0.005.

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

    FREM1 regulates Timp1 transcription downstream of PDGFCC, through a mechanism that is dependent on PDGFRα, PI3K and MAPK activation. (A) qRT-PCR analysis of Timp1 mRNA levels in WT (black bars) and bat mutant (white bars) MEFs following PDGFCC stimulation for the indicated time points. Data was obtained from four independent experiments from at least three different cell lines for each genotype, and presented as fold increase relative to unstimulated WT cells. (B) Alterations in phosphorylation of AKT and ERK1/2 precede changes in Timp1 expression. (C) TIMP1 protein secretion is reduced in stimulated FREM1 bat mutant MEF cultures. (D) WT cells were pre-treated with either the PI3K inhibitor LY294002, AKT1/2 inhibitor, MAPK inhibitor UO126 or DMSO vehicle control prior to PDGFCC stimulation and analysed by immunoblotting with anti-phospho-AKT, anti-phospho-ERK1/2, total AKT or total ERK1/2. (E) qRT-PCR for Timp1 mRNA was performed on the same cells. Levels of Timp1 mRNA were presented as a fold increase relative to unstimulated cells. Experiments were performed three times using three different cell lines. (F) E13.5 WT and bat embryo head skin sections stained for COL1 (green), keratin 14 (K14; red) and nuclear dye DAPI (blue). Error bars represent s.e.m.; *P<0.05, **P<0.01.

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

    Summary model of FREM1 bat mutant basement membrane fragility. (Left) In WT mice, FREM1 forms a stabilising complex (1) with FRAS/FREM proteins to cross-link the lamina densa with the underlying dermis. Additionally, FREM1 binds keratinocyte-secreted PDGFC, which is presented to PDGFRα in the adjacent dermal fibroblasts, potentiating PDGFRα signalling and promoting ECM modelling events, including TIMP1 upregulation and COL1 deposition (2). (Right) In bat mice, FREM1 mutation removes the structural cross-link of the FRAS/FREM complex (1), but also reduces PDGFRα signalling, leading to lowered TIMP1 and diminished COL1 deposition (2), thereby further weakening part of the foundation of the basement membrane.

Previous ArticleNext Article
Back to top
Previous ArticleNext Article

This Issue

RSSRSS

 Download PDF

Email

Thank you for your interest in spreading the word on Disease Models & Mechanisms.

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.
Regulation of PDGFC signalling and extracellular matrix composition by FREM1 in mice
(Your Name) has sent you a message from Disease Models & Mechanisms
(Your Name) thought you would like to see the Disease Models & Mechanisms 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
Regulation of PDGFC signalling and extracellular matrix composition by FREM1 in mice
Fenny Wiradjaja, Denny L. Cottle, Lynelle Jones, Ian Smyth
Disease Models & Mechanisms 2013 6: 1426-1433; doi: 10.1242/dmm.013748
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Research Article
Regulation of PDGFC signalling and extracellular matrix composition by FREM1 in mice
Fenny Wiradjaja, Denny L. Cottle, Lynelle Jones, Ian Smyth
Disease Models & Mechanisms 2013 6: 1426-1433; doi: 10.1242/dmm.013748

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
    • SUMMARY
    • INTRODUCTION
    • RESULTS
    • DISCUSSION
    • MATERIALS AND METHODS
    • Acknowledgements
    • Footnotes
    • REFERENCES
  • Figures & tables
  • Info & metrics
  • PDF

Related articles

Cited by...

More in this TOC section

  • Altered cytoskeletal arrangement in induced pluripotent stem cells (iPSCs) and motor neurons from patients with riboflavin transporter deficiency
  • Genetic background modifies vulnerability to glaucoma related phenotypes in Lmx1b mutant mice
  • Heterogeneity in clone dynamics within and adjacent to intestinal tumours identified by Dre-mediated lineage tracing
Show more RESEARCH ARTICLE

Similar articles

Other journals from The Company of Biologists

Development

Journal of Cell Science

Journal of Experimental Biology

Biology Open

Advertisement

DMM and COVID-19

We are aware that the COVID-19 pandemic is having an unprecedented impact on researchers worldwide. The Editors of all The Company of Biologists’ journals have been considering ways in which we can alleviate concerns that members of our community may have around publishing activities during this time. Read about the actions we are taking at this time.

Please don’t hesitate to contact the Editorial Office if you have any questions or concerns.


Professor Elizabeth Patton appointed as DMM’s next Editor-in-Chief

We are pleased to announce that The Company of Biologists directors have appointed Professor Elizabeth Patton as DMM's new Editor-in-Chief. As Paresh Vyas writes in his Editorial, Liz ‘brings vitality and a passion for the remit of DMM, and is deeply embedded in the community.’


Did you know DMM Conference Travel Grants can be used for online meetings?

With travel restrictions still in place, we want to continue supporting early-career researchers in their careers. DMM’s Conference Travel Grants can now be used to attend virtual and online scientific meetings, workshops, conferences and training courses.

The current application round closes on 8 February 2021 – find out more.


Identification of MYOM2 as a candidate gene in hypertrophic cardiomyopathy and Tetralogy of Fallot, and its functional evaluation in the Drosophila heart

Research from Silke Sperling and colleagues uses Drosophila to identify MYOM2 as a candidate gene in congenital heart malformations in this issue’s Editor’s choice.


C. elegans as a disease model

A new Research article from Doyle et al., models spinal muscular atrophy in C. elegans to show that that targeting therapies to muscle cells is more effective than neuronal delivery. Find more research using C. elegans as a disease model in our latest subject collection.


Call for papers – The RAS Pathway: Diseases, Therapeutics and Beyond

Our upcoming special issue is now welcoming submissions until 1 April 2021. Guest-edited by Donita Brady (Perelman School of Medicine at the University of Pennsylvania, USA) and Arvin Dar (Icahn School of Medicine at Mount Sinai, USA), the issue will focus on the targeting the RAS pathway. Find out more about the issue and how to submit your manuscript.


Interview – Kim Landry-Truchon and Nicolas Houde

In an interview, first authors Kim Landry-Truchon and Nicolas Houde discuss their mouse model of the early stages of pleuropulmonary blastoma, reflecting on the implications of their work and the future of their field.

Articles

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

About us

  • About DMM
  • 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 DMM
  • Advertising
  • Feedback

Twitter   YouTube   LinkedIn

© 2021   The Company of Biologists Ltd   Registered Charity 277992