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Cellular and Molecular Neurobiology

, Volume 38, Issue 8, pp 1557–1563 | Cite as

A Novel Microfluidic Device-Based Neurite Outgrowth Inhibition Assay Reveals the Neurite Outgrowth-Promoting Activity of Tropomyosin Tpm3.1 in Hippocampal Neurons

  • Holly Stefen
  • Amin Hassanzadeh-Barforoushi
  • Merryn Brettle
  • Sandra Fok
  • Alexandra K. Suchowerska
  • Nicodemus Tedla
  • Tracie Barber
  • Majid Ebrahimi Warkiani
  • Thomas Fath
Brief Communication

Abstract

Overcoming neurite inhibition is integral for restoring neuronal connectivity after CNS injury. Actin dynamics are critical for neurite growth cone formation and extension. The tropomyosin family of proteins is a regarded as master regulator of actin dynamics. This study investigates tropomyosin isoform 3.1 (Tpm3.1) as a potential candidate for overcoming an inhibitory substrate, as it is known to influence neurite branching and outgrowth. We designed a microfluidic device that enables neurons to be grown adjacent to an inhibitory substrate, Nogo-66. Results show that neurons, overexpressing hTpm3.1, have an increased propensity to overcome Nogo-66 inhibition. We propose Tpm3.1 as a potential target for promoting neurite growth in an inhibitory environment in the central nervous system.

Keywords

Tropomyosins Neurite outgrowth inhibition Microfluidic systems NogoA 

Notes

Acknowledgements

This work was supported by Project Grant APP1083209 from the Australian National Health and Medical Research Council (NHMRC) (T.F.) and Discovery Project Grant DP180101473 from the Australian Research Council (ARC) (T.F.). We thank Tamara Tomanić (UNSW Sydney) for her constructive feedback and critical reading of the manuscript. This work was performed (in part) at the NSW and South Australian node of the Australian National Fabrication Facility under the National Collaborative Research Infrastructure Strategy to provide nano- and microfabrication facilities for Australia’s researchers. M.E.W. would like to acknowledge the support of the Australian Research Council through Discovery Project Grants (DP170103704 and DP180103003) and the National Health and Medical Research Council through the Career Development Fellowship (APP1143377).

Author Contributions

TF, MEW and TB supervised the project. TF, HS, AH-B and SF designed the research. HS, AH-B, SF and MB performed the research and analysed the data. TF, HS, AH-B and SF wrote the paper. TF, HS, AH-B, AKS, MB and NT edited the paper. All authors read and approved the final manuscript.

Compliance with Ethical Standards

Conflict of interest

The authors declare that the research was conducted in the absence of any commercial, financial or non-financial relationships that could be construed as a potential conflict of interest.

Ethical Approval

All procedures were conducted in accordance with the Australian Code of Practice for the Care and Use of Animals for Scientific Purposes and were approved by the University of New South Wales Animal Care and Ethics Committee.

Supplementary material

10571_2018_620_MOESM1_ESM.docx (23 kb)
Supplementary material 1 (DOCX 22 KB)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Neurodegenerative and Repair Unit, School of Medical ScienceUNSW SydneySydneyAustralia
  2. 2.Neuron Culture Core Facility (NCCF)University of New South WalesSydneyAustralia
  3. 3.School of Mechanical and Manufacturing EngineeringUniversity of New South WalesSydneyAustralia
  4. 4.Cancer DivisionGarvan Institute of Medical Research/The Kinghorn Cancer CentreSydneyAustralia
  5. 5.Inflammation Research, School of Medical SciencesUniversity of New South WalesSydneyAustralia
  6. 6.School of Biomedical EngineeringUniversity of Technology SydneySydneyAustralia
  7. 7.Institute of Molecular MedicineSechenov First Moscow State UniversityMoscowRussia
  8. 8.Faculty of Medicine and Health Sciences, Dementia Research CentreMacquarie UniversitySydneyAustralia

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