Expression and knockdown of zebrafish folliculin suggests requirement for embryonic brain morphogenesis

Emma J Kenyon, Monique N H Luijten, Harmeet Gill, Nan Li, Matthew Rawlings, James C Bull, Yavor Hadzhiev, Maurice A M van Steensel, Eamonn Maher, Ferenc Mueller

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    Abstract

    Background: Birt-Hogg-Dubé syndrome (BHD) is a dominantly inherited familial cancer syndrome characterised by the development of benign skin fibrofolliculomas, multiple lung and kidney cysts, spontaneous pneumothorax and susceptibility to renal cell carcinoma. BHD is caused by mutations in the gene encoding Folliculin (FLCN). Little is known about what FLCN does in a healthy individual and how best to treat those with BHD. As a first approach to developing a vertebrate model for BHD we aimed to identify the temporal and spatial expression of flcn transcripts in the developing zebrafish embryo. To gain insights into the function of flcn in a whole organism system we generated a loss of function model of flcn by the use of morpholino knockdown in zebrafish.

    Results: flcn is expressed broadly and upregulated in the fin bud, somites, eye and proliferative regions of the brain of the Long-pec stage zebrafish embryos. Together with knockdown phenotypes, expression analysis suggest involvement of flcn in zebrafish embryonic brain development. We have utilised the zFucci system, an in vivo, whole organism cell cycle assay to study the potential role of flcn in brain development. We found that at the 18 somite stage there was a significant drop in cells in the S-M phase of the cell cycle in flcn morpholino injected embryos with a corresponding increase of cells in the G1 phase. This was particularly evident in the brain, retina and somites of the embryo. Timelapse analysis of the head region of flcn morpholino injected and mismatch control embryos shows the temporal dynamics of cell cycle misregulation during development.

    Conclusions: In conclusion we show that zebrafish flcn is expressed in a non-uniform manner and is likely required for the maintenance of correct cell cycle regulation during embryonic development. We demonstrate the utilisation of the zFucci system in testing the role of flcn in cell proliferation and suggest a function for flcn in regulating cell proliferation in vertebrate embryonic brain development.

    Original languageEnglish
    Article number23
    Pages (from-to)1-14
    Number of pages14
    JournalBMC Developmental Biology
    Volume16
    DOIs
    Publication statusPublished - 8 Jul 2016

    Fingerprint

    Estrone
    Zebrafish
    Morphogenesis
    Embryonic Structures
    Morpholinos
    Somites
    Cell Cycle
    Brain
    Embryonic Development
    Vertebrates
    Cell Proliferation
    G1 Phase
    Pneumothorax
    S Phase
    Renal Cell Carcinoma
    Cell Division
    Retina
    Cysts
    Head
    Maintenance

    Keywords

    • Birt-Hogg-Dube syndrome (BHD)
    • Zebrafish

    Cite this

    Kenyon, E. J., Luijten, M. N. H., Gill, H., Li, N., Rawlings, M., Bull, J. C., ... Mueller, F. (2016). Expression and knockdown of zebrafish folliculin suggests requirement for embryonic brain morphogenesis. BMC Developmental Biology, 16, 1-14. [23]. https://doi.org/10.1186/s12861-016-0119-8
    Kenyon, Emma J ; Luijten, Monique N H ; Gill, Harmeet ; Li, Nan ; Rawlings, Matthew ; Bull, James C ; Hadzhiev, Yavor ; van Steensel, Maurice A M ; Maher, Eamonn ; Mueller, Ferenc. / Expression and knockdown of zebrafish folliculin suggests requirement for embryonic brain morphogenesis. In: BMC Developmental Biology. 2016 ; Vol. 16. pp. 1-14.
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    title = "Expression and knockdown of zebrafish folliculin suggests requirement for embryonic brain morphogenesis",
    abstract = "Background: Birt-Hogg-Dub{\'e} syndrome (BHD) is a dominantly inherited familial cancer syndrome characterised by the development of benign skin fibrofolliculomas, multiple lung and kidney cysts, spontaneous pneumothorax and susceptibility to renal cell carcinoma. BHD is caused by mutations in the gene encoding Folliculin (FLCN). Little is known about what FLCN does in a healthy individual and how best to treat those with BHD. As a first approach to developing a vertebrate model for BHD we aimed to identify the temporal and spatial expression of flcn transcripts in the developing zebrafish embryo. To gain insights into the function of flcn in a whole organism system we generated a loss of function model of flcn by the use of morpholino knockdown in zebrafish.Results: flcn is expressed broadly and upregulated in the fin bud, somites, eye and proliferative regions of the brain of the Long-pec stage zebrafish embryos. Together with knockdown phenotypes, expression analysis suggest involvement of flcn in zebrafish embryonic brain development. We have utilised the zFucci system, an in vivo, whole organism cell cycle assay to study the potential role of flcn in brain development. We found that at the 18 somite stage there was a significant drop in cells in the S-M phase of the cell cycle in flcn morpholino injected embryos with a corresponding increase of cells in the G1 phase. This was particularly evident in the brain, retina and somites of the embryo. Timelapse analysis of the head region of flcn morpholino injected and mismatch control embryos shows the temporal dynamics of cell cycle misregulation during development.Conclusions: In conclusion we show that zebrafish flcn is expressed in a non-uniform manner and is likely required for the maintenance of correct cell cycle regulation during embryonic development. We demonstrate the utilisation of the zFucci system in testing the role of flcn in cell proliferation and suggest a function for flcn in regulating cell proliferation in vertebrate embryonic brain development.",
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    Kenyon, EJ, Luijten, MNH, Gill, H, Li, N, Rawlings, M, Bull, JC, Hadzhiev, Y, van Steensel, MAM, Maher, E & Mueller, F 2016, 'Expression and knockdown of zebrafish folliculin suggests requirement for embryonic brain morphogenesis', BMC Developmental Biology, vol. 16, 23, pp. 1-14. https://doi.org/10.1186/s12861-016-0119-8

    Expression and knockdown of zebrafish folliculin suggests requirement for embryonic brain morphogenesis. / Kenyon, Emma J; Luijten, Monique N H; Gill, Harmeet; Li, Nan; Rawlings, Matthew; Bull, James C; Hadzhiev, Yavor; van Steensel, Maurice A M; Maher, Eamonn; Mueller, Ferenc.

    In: BMC Developmental Biology, Vol. 16, 23, 08.07.2016, p. 1-14.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Expression and knockdown of zebrafish folliculin suggests requirement for embryonic brain morphogenesis

    AU - Kenyon, Emma J

    AU - Luijten, Monique N H

    AU - Gill, Harmeet

    AU - Li, Nan

    AU - Rawlings, Matthew

    AU - Bull, James C

    AU - Hadzhiev, Yavor

    AU - van Steensel, Maurice A M

    AU - Maher, Eamonn

    AU - Mueller, Ferenc

    PY - 2016/7/8

    Y1 - 2016/7/8

    N2 - Background: Birt-Hogg-Dubé syndrome (BHD) is a dominantly inherited familial cancer syndrome characterised by the development of benign skin fibrofolliculomas, multiple lung and kidney cysts, spontaneous pneumothorax and susceptibility to renal cell carcinoma. BHD is caused by mutations in the gene encoding Folliculin (FLCN). Little is known about what FLCN does in a healthy individual and how best to treat those with BHD. As a first approach to developing a vertebrate model for BHD we aimed to identify the temporal and spatial expression of flcn transcripts in the developing zebrafish embryo. To gain insights into the function of flcn in a whole organism system we generated a loss of function model of flcn by the use of morpholino knockdown in zebrafish.Results: flcn is expressed broadly and upregulated in the fin bud, somites, eye and proliferative regions of the brain of the Long-pec stage zebrafish embryos. Together with knockdown phenotypes, expression analysis suggest involvement of flcn in zebrafish embryonic brain development. We have utilised the zFucci system, an in vivo, whole organism cell cycle assay to study the potential role of flcn in brain development. We found that at the 18 somite stage there was a significant drop in cells in the S-M phase of the cell cycle in flcn morpholino injected embryos with a corresponding increase of cells in the G1 phase. This was particularly evident in the brain, retina and somites of the embryo. Timelapse analysis of the head region of flcn morpholino injected and mismatch control embryos shows the temporal dynamics of cell cycle misregulation during development.Conclusions: In conclusion we show that zebrafish flcn is expressed in a non-uniform manner and is likely required for the maintenance of correct cell cycle regulation during embryonic development. We demonstrate the utilisation of the zFucci system in testing the role of flcn in cell proliferation and suggest a function for flcn in regulating cell proliferation in vertebrate embryonic brain development.

    AB - Background: Birt-Hogg-Dubé syndrome (BHD) is a dominantly inherited familial cancer syndrome characterised by the development of benign skin fibrofolliculomas, multiple lung and kidney cysts, spontaneous pneumothorax and susceptibility to renal cell carcinoma. BHD is caused by mutations in the gene encoding Folliculin (FLCN). Little is known about what FLCN does in a healthy individual and how best to treat those with BHD. As a first approach to developing a vertebrate model for BHD we aimed to identify the temporal and spatial expression of flcn transcripts in the developing zebrafish embryo. To gain insights into the function of flcn in a whole organism system we generated a loss of function model of flcn by the use of morpholino knockdown in zebrafish.Results: flcn is expressed broadly and upregulated in the fin bud, somites, eye and proliferative regions of the brain of the Long-pec stage zebrafish embryos. Together with knockdown phenotypes, expression analysis suggest involvement of flcn in zebrafish embryonic brain development. We have utilised the zFucci system, an in vivo, whole organism cell cycle assay to study the potential role of flcn in brain development. We found that at the 18 somite stage there was a significant drop in cells in the S-M phase of the cell cycle in flcn morpholino injected embryos with a corresponding increase of cells in the G1 phase. This was particularly evident in the brain, retina and somites of the embryo. Timelapse analysis of the head region of flcn morpholino injected and mismatch control embryos shows the temporal dynamics of cell cycle misregulation during development.Conclusions: In conclusion we show that zebrafish flcn is expressed in a non-uniform manner and is likely required for the maintenance of correct cell cycle regulation during embryonic development. We demonstrate the utilisation of the zFucci system in testing the role of flcn in cell proliferation and suggest a function for flcn in regulating cell proliferation in vertebrate embryonic brain development.

    KW - Birt-Hogg-Dube syndrome (BHD)

    KW - Zebrafish

    U2 - 10.1186/s12861-016-0119-8

    DO - 10.1186/s12861-016-0119-8

    M3 - Article

    VL - 16

    SP - 1

    EP - 14

    JO - BMC Developmental Biology

    JF - BMC Developmental Biology

    SN - 1471-213X

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