Molecular Neurobiology

, Volume 55, Issue 5, pp 4051–4067 | Cite as

A BRCA1-Dependent DNA Damage Response in the Regenerating Adult Peripheral Nerve Milieu

  • Anand Krishnan
  • Kaylynn Purdy
  • Ambika Chandrasekhar
  • Jose Martinez
  • Chu Cheng
  • Douglas W. Zochodne
Article
  • 248 Downloads

Abstract

It is not generally appreciated that DNA repair machinery has a critical role in the remodeling of neurons that adopt a regenerative phenotype. We identified that breast cancer 1 (BRCA1)-dependent DNA activity, previously well known to repair cancer cells, is active in adult peripheral neurons and Schwann cells during their injury and regeneration response. Temporary or partial loss of BRCA1 or blockade of its intraneuronal nuclear entry impaired outgrowth in neurons in vitro and impacted nerve regeneration and functional recovery in vivo. We found that distal axonal injury triggered a BRCA1-dependent DNA damage response (DDR) signal in neuronal soma. BRCA1 also supported an enabling transcriptional program of injured neurons and supporting Schwann cells. Our findings indicate that BRCA1 offers prominent functional roles in neurons and glial cells including key support for their physical and molecular integrity. Since BRCA1 mutations are common in humans, this function of BRCA1 in peripheral neurons and their glial partners warrants attention.

Keywords

BRCA1 Neuron DRG Schwann cells Peripheral nerve Regeneration 

Notes

Acknowledgements

The work was supported by operating grants from the Canadian Institutes of Health Research (CIHR; FRN15686 and 184584) and the Canadian Diabetes Association (CDA; OG-3-12-3669) as well as support from the University of Alberta Hospital Foundation, the Department of Medicine and the Division of Neurology, Faculty of Medicine and Dentistry, University of Alberta. AK was supported by an Alberta Innovates Health Solutions (AI-HS) postdoctoral fellowship. The authors thank Dr. Jan Van Minnen for the gift of SCs used in this work. BRCA1-floxed mice breeder pairs were obtained from the National Cancer Institute (NCI) mouse repository.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no competing interests.

Supplementary material

12035_2017_574_Fig12_ESM.gif (52 kb)
Fig. S1

BRCA1 expression in satellite glial cells and primary sensory neurons. A&B) wider field view of BRCA1 immunostaining (green) in injury-conditioned DRGs shows its expression in perineuronal satellite cells (yellow arrows) and neuronal nuclei (white arrows) (A; scale bar, 50 μm, B; scale bar, 200 μm). C) Nuclear accumulation of BRCA1 in primary sensory neurons coincides with enhanced neurite outgrowth. The neuron enlarged in a’ has comparatively less nuclear BRCA1 content (yellow arrow) compared to the neuron in b’ while the latter has comparatively higher outgrowth (scale bar 50 μm). The corresponding original neurons are shown using white arrows. (GIF 51 kb)

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High resolution image (TIFF 2512 kb)
12035_2017_574_Fig13_ESM.gif (80 kb)
Fig. S2

Validation of the impact of BRCA1NLS-TAT on nuclear clustering of BRCA1. Immunostaining of BRCA1 in primary sensory neurons treated with cisplatin shows nuclear clustering of BRCA1 (white arrows in upper panel). 24 h pretreatment with 10 μM BRCA1NLS-TAT reduces the nuclear clustering (white arrows in lower panel) (scale bar 50 μm). Quantification of the data from corresponding single culture wells is given in the graph. (GIF 79 kb)

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High resolution image (TIFF 12786 kb)
12035_2017_574_Fig14_ESM.gif (17 kb)
Fig. S3

Neurite outgrowth in floxed-BRCA1 sensory neurons treated with either Ad-GFP or Ad-GFP-2A-iCre. The figures show a merged view of NF200 (red), GFP (green) and DAPI (blue). Expression of GFP in Ad-GFP-2A-iCre treated neurons indicates successful viral particle infection and BRCA1 knockdown efficiency. (GIF 17 kb)

12035_2017_574_MOESM3_ESM.tif (2 mb)
High resolution image (TIFF 2006 kb)
12035_2017_574_Fig15_ESM.jpg (42 kb)
Fig. S4

BRCA1 deficiency alters transcription in sensory neurons and SCs. Pie charts generated from altered list of mRNAs after BRCA1 knockdown in sensory neurons (A) and SCs (B) show classification of altered molecules based on their participating functions. The results indicate that BRCA1 deficiency mainly altered the expression of molecules involved with DNA repair, neurite outgrowth and apoptosis in sensory neurons and molecules involved with cell cycle, DNA repair and apoptosis in SCs [see Table S2 for fold change and ‘p’ values]. (JPG 41.7 kb)

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High resolution image (TIFF 1.78 mb)
12035_2017_574_MOESM5_ESM.docx (16 kb)
Table S1 (DOCX 16 kb)
12035_2017_574_MOESM6_ESM.doc (192 kb)
Table S2 (DOC 192 kb)

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Anand Krishnan
    • 1
    • 2
    • 3
    • 4
  • Kaylynn Purdy
    • 1
    • 2
    • 3
    • 4
  • Ambika Chandrasekhar
    • 1
    • 2
    • 3
  • Jose Martinez
    • 4
  • Chu Cheng
    • 4
  • Douglas W. Zochodne
    • 1
    • 2
    • 3
    • 4
  1. 1.Division of Neurology, Department of MedicineUniversity of AlbertaEdmontonCanada
  2. 2.Neuroscience and Mental Health InstituteUniversity of AlbertaEdmontonCanada
  3. 3.Alberta Diabetes InstituteUniversity of AlbertaEdmontonCanada
  4. 4.Hotchkiss Brain InstituteUniversity of CalgaryCalgaryCanada

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