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Effects of GDF5 overexpression on embryonic rat dopaminergic neurones in vitro and in vivo

  • Basic Neurosciences, Genetics and Immunology - Original Articles
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Abstract

Transplantation of embryonic dopaminergic neurones has shown promise for the treatment of Parkinson’s disease (PD), but this approach is limited by the poor survival of the transplanted cells. Exogenous dopaminergic neurotrophic factors such as growth/differentiation factor 5 (GDF5) have been found to enhance the survival of transplanted dopaminergic neurones. However, this approach is limited by the rapid degradation of such factors in vivo; thus, methods for long-term delivery of these factors are under investigation. The present study shows, using optimised lipid-mediated transfection procedures, that overexpression of GDF5 significantly improves the survival of dopaminergic neurones in cultures of embryonic day (E) 13 rat ventral mesencephalon (VM) and protects them against 6-hydroxydopamine (6-OHDA)-induced toxicity. In another experiment, E13 VM cells were transfected with GDF5 after 1 day in vitro (DIV), then transplanted into 6-OHDA-lesioned adult rat striata after 2 DIV. The survival of these E13 VM dopaminergic neurones after transfection and transplantation was as least as high as that of freshly dissected E14 VM dopaminergic neurones, demonstrating that transfection was not detrimental to these cells. Furthermore, GDF5-overexpressing E13 VM transplants significantly reduced amphetamine-induced rotational asymmetry in the lesioned rats. This study shows that lipid-mediated transfection in vitro prior to transplantation is a valid approach for the introduction of neurotrophic proteins such as GDF5, as well as lending further support to the potential use of GDF5 in neuroprotective therapy for PD.

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Abbreviations

6-OHDA:

6-Hydroxydopamine

DIV:

Day in vitro

E:

Embryonic day

GDF5:

Growth/differentiation factor 5

GDNF:

Glial cell line-derived neurotrophic factor

GFAP:

Glial fibrillary acidic protein

PD:

Parkinson’s disease

PI:

Propidium iodide

TGF-β:

Transforming growth factor-β

TH:

Tyrosine hydroxylase

VM:

Ventral mesencephalon

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Acknowledgments

This work was funded by the Health Research Board of Ireland. The authors wish to thank Dr Jens Pohl and Dr Michael Hanke of Biopharm GmbH, Germany, for the generous gifts of pSGDF5 plasmid and aMP5 antibody. We also wish to acknowledge Ms Bereniece Riedewald for her assistance with the preparation of the figures and Dr Gerard O’Keeffe for critical reading of the manuscript.

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Authors and Affiliations

  1. Department of Neuroscience/Anatomy, Biosciences Research Institute, University College Cork, College Road, Cork, Ireland

    David B. O’Sullivan & Aideen M. Sullivan

  2. Department of Physiology, Biosciences Research Institute, University College Cork, College Road, Cork, Ireland

    Patrick T. Harrison

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  1. David B. O’Sullivan
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Correspondence to Aideen M. Sullivan.

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O’Sullivan, D.B., Harrison, P.T. & Sullivan, A.M. Effects of GDF5 overexpression on embryonic rat dopaminergic neurones in vitro and in vivo. J Neural Transm 117, 559–572 (2010). https://doi.org/10.1007/s00702-010-0392-9

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