C9ORF72 expression and cellular localization over mouse development

Rachel A. K. Atkinson, Carmen M. Fernandez-Martos, Julie D. Atkin, James C. Vickers, Anna E. King

    Research output: Contribution to journalArticleResearchpeer-review

    Abstract

    INTRODUCTION: A majority of familial frontotemporal lobar dementia and amyotrophic lateral sclerosis cases are associated with a large repeat expansion in a non-coding region of the C9ORF72 gene. Currently, little is known about the normal function and the expression pattern of the C9ORF72 protein. The aims of this study were to characterize the expression pattern and cellular localization of the three reported mouse isoforms of C9orf72, over a developmental time-course in primary cultured cortical neurons and brain tissue from C57BL/6 mice. RESULTS: We demonstrated that the different isoforms of C9ORF72 at the mRNA and protein level undergo alterations in expression during development and into adulthood. Cellular fractionation and immunofluorescence demonstrated that levels of nuclear and cytoplasmic expression of isoforms changed significantly over the time course. Additionally, immunofluorescence studies showed C9ORF72 labeling as puncta throughout neurons, extending beyond the microtubule cytoskeleton into actin-rich structures such as filopodia and growth cones. Finally, synaptosome preparations demonstrated the presence of C9ORF72 isoform 1 in synaptic-rich fractions from adult mouse brain. CONCLUSION: In summary, the presence of C9ORF72 as puncta and within synaptic-rich fractions may indicate involvement at the synapse and differential expression of isoforms in nuclei and cytoplasm may suggest distinct roles for the isoforms. Determining the physiological role of C9ORF72 protein may help to determine the role it plays in disease.

    LanguageEnglish
    Article number59
    Pages1-11
    Number of pages11
    JournalActa neuropathologica communications
    Volume3
    DOIs
    Publication statusPublished - 25 Sep 2015

    Fingerprint

    Protein Isoforms
    Fluorescent Antibody Technique
    Neurons
    Growth Cones
    Proteins
    Pseudopodia
    Synaptosomes
    Brain
    Actin Cytoskeleton
    Inbred C57BL Mouse
    Microtubules
    Synapses
    Cytoplasm
    Messenger RNA
    Genes

    Bibliographical note

    Copyright the Author(s) 2015. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

    Keywords

    • C9ORF72
    • FTLD
    • ALS

    Cite this

    Atkinson, Rachel A. K. ; Fernandez-Martos, Carmen M. ; Atkin, Julie D. ; Vickers, James C. ; King, Anna E. / C9ORF72 expression and cellular localization over mouse development. In: Acta neuropathologica communications. 2015 ; Vol. 3. pp. 1-11.
    @article{273ca2c724fd43a7bb54a13d36b7e66f,
    title = "C9ORF72 expression and cellular localization over mouse development",
    abstract = "INTRODUCTION: A majority of familial frontotemporal lobar dementia and amyotrophic lateral sclerosis cases are associated with a large repeat expansion in a non-coding region of the C9ORF72 gene. Currently, little is known about the normal function and the expression pattern of the C9ORF72 protein. The aims of this study were to characterize the expression pattern and cellular localization of the three reported mouse isoforms of C9orf72, over a developmental time-course in primary cultured cortical neurons and brain tissue from C57BL/6 mice. RESULTS: We demonstrated that the different isoforms of C9ORF72 at the mRNA and protein level undergo alterations in expression during development and into adulthood. Cellular fractionation and immunofluorescence demonstrated that levels of nuclear and cytoplasmic expression of isoforms changed significantly over the time course. Additionally, immunofluorescence studies showed C9ORF72 labeling as puncta throughout neurons, extending beyond the microtubule cytoskeleton into actin-rich structures such as filopodia and growth cones. Finally, synaptosome preparations demonstrated the presence of C9ORF72 isoform 1 in synaptic-rich fractions from adult mouse brain. CONCLUSION: In summary, the presence of C9ORF72 as puncta and within synaptic-rich fractions may indicate involvement at the synapse and differential expression of isoforms in nuclei and cytoplasm may suggest distinct roles for the isoforms. Determining the physiological role of C9ORF72 protein may help to determine the role it plays in disease.",
    keywords = "C9ORF72, FTLD, ALS",
    author = "Atkinson, {Rachel A. K.} and Fernandez-Martos, {Carmen M.} and Atkin, {Julie D.} and Vickers, {James C.} and King, {Anna E.}",
    note = "Copyright the Author(s) 2015. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.",
    year = "2015",
    month = "9",
    day = "25",
    doi = "10.1186/s40478-015-0238-7",
    language = "English",
    volume = "3",
    pages = "1--11",
    journal = "Acta neuropathologica communications",
    issn = "2051-5960",
    publisher = "Springer, Springer Nature",

    }

    C9ORF72 expression and cellular localization over mouse development. / Atkinson, Rachel A. K.; Fernandez-Martos, Carmen M.; Atkin, Julie D.; Vickers, James C.; King, Anna E.

    In: Acta neuropathologica communications, Vol. 3, 59, 25.09.2015, p. 1-11.

    Research output: Contribution to journalArticleResearchpeer-review

    TY - JOUR

    T1 - C9ORF72 expression and cellular localization over mouse development

    AU - Atkinson, Rachel A. K.

    AU - Fernandez-Martos, Carmen M.

    AU - Atkin, Julie D.

    AU - Vickers, James C.

    AU - King, Anna E.

    N1 - Copyright the Author(s) 2015. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

    PY - 2015/9/25

    Y1 - 2015/9/25

    N2 - INTRODUCTION: A majority of familial frontotemporal lobar dementia and amyotrophic lateral sclerosis cases are associated with a large repeat expansion in a non-coding region of the C9ORF72 gene. Currently, little is known about the normal function and the expression pattern of the C9ORF72 protein. The aims of this study were to characterize the expression pattern and cellular localization of the three reported mouse isoforms of C9orf72, over a developmental time-course in primary cultured cortical neurons and brain tissue from C57BL/6 mice. RESULTS: We demonstrated that the different isoforms of C9ORF72 at the mRNA and protein level undergo alterations in expression during development and into adulthood. Cellular fractionation and immunofluorescence demonstrated that levels of nuclear and cytoplasmic expression of isoforms changed significantly over the time course. Additionally, immunofluorescence studies showed C9ORF72 labeling as puncta throughout neurons, extending beyond the microtubule cytoskeleton into actin-rich structures such as filopodia and growth cones. Finally, synaptosome preparations demonstrated the presence of C9ORF72 isoform 1 in synaptic-rich fractions from adult mouse brain. CONCLUSION: In summary, the presence of C9ORF72 as puncta and within synaptic-rich fractions may indicate involvement at the synapse and differential expression of isoforms in nuclei and cytoplasm may suggest distinct roles for the isoforms. Determining the physiological role of C9ORF72 protein may help to determine the role it plays in disease.

    AB - INTRODUCTION: A majority of familial frontotemporal lobar dementia and amyotrophic lateral sclerosis cases are associated with a large repeat expansion in a non-coding region of the C9ORF72 gene. Currently, little is known about the normal function and the expression pattern of the C9ORF72 protein. The aims of this study were to characterize the expression pattern and cellular localization of the three reported mouse isoforms of C9orf72, over a developmental time-course in primary cultured cortical neurons and brain tissue from C57BL/6 mice. RESULTS: We demonstrated that the different isoforms of C9ORF72 at the mRNA and protein level undergo alterations in expression during development and into adulthood. Cellular fractionation and immunofluorescence demonstrated that levels of nuclear and cytoplasmic expression of isoforms changed significantly over the time course. Additionally, immunofluorescence studies showed C9ORF72 labeling as puncta throughout neurons, extending beyond the microtubule cytoskeleton into actin-rich structures such as filopodia and growth cones. Finally, synaptosome preparations demonstrated the presence of C9ORF72 isoform 1 in synaptic-rich fractions from adult mouse brain. CONCLUSION: In summary, the presence of C9ORF72 as puncta and within synaptic-rich fractions may indicate involvement at the synapse and differential expression of isoforms in nuclei and cytoplasm may suggest distinct roles for the isoforms. Determining the physiological role of C9ORF72 protein may help to determine the role it plays in disease.

    KW - C9ORF72

    KW - FTLD

    KW - ALS

    UR - http://www.scopus.com/inward/record.url?scp=84983477356&partnerID=8YFLogxK

    U2 - 10.1186/s40478-015-0238-7

    DO - 10.1186/s40478-015-0238-7

    M3 - Article

    VL - 3

    SP - 1

    EP - 11

    JO - Acta neuropathologica communications

    T2 - Acta neuropathologica communications

    JF - Acta neuropathologica communications

    SN - 2051-5960

    M1 - 59

    ER -