Journal of Molecular Neuroscience

, Volume 46, Issue 3, pp 654–665

Optimized Quantities of GDNF Overexpressed by Engineered Astrocytes Are Critical for Protection of Neuroblastoma Cells Against 6-OHDA Toxicity

Authors

  • Roya Safi
    • Molecular Genetics GroupNational Institute of Genetic Engineering and Biotechnology (NIGEB)
    • Molecular Genetics GroupNational Institute of Genetic Engineering and Biotechnology (NIGEB)
    • Neuroscience Research Center (NRC)Shahid Beheshti University of Medical Sciences
  • Yasin Panahi
    • Molecular Genetics GroupNational Institute of Genetic Engineering and Biotechnology (NIGEB)
  • Nader Maghsoudi
    • Neuroscience Research Center (NRC)Shahid Beheshti University of Medical Sciences
  • Mohammad Zaefizadeh
    • Molecular Genetics GroupNational Institute of Genetic Engineering and Biotechnology (NIGEB)
  • Ehsan Gharib
    • Molecular Genetics GroupNational Institute of Genetic Engineering and Biotechnology (NIGEB)
Article

DOI: 10.1007/s12031-011-9654-8

Cite this article as:
Safi, R., Gardaneh, M., Panahi, Y. et al. J Mol Neurosci (2012) 46: 654. doi:10.1007/s12031-011-9654-8

Abstract

Optimized levels of glial cell line-derived neurotrophic factor glial cell line-derived neurotrophic factor (GDNF) are critical for protection of dopaminergic neurons against parkinsonian cell death. Recombinant lentiviruses harboring GDNF coding sequence were constructed and used to infect astrocytoma cell line 1321N1. The infected astrocytes overexpressed GDNF mRNA and secreted an average of 2.2 ng/mL recombinant protein as tested in both 2 and 16 weeks post-infection. Serial dilutions of GDNF-enriched conditioned medium from infected astrocytes added to growing neuroblastoma cell line SK-N-MC resulted in commensurate resistance against 6-OHDA toxicity. SK-N-MC cell survival rate rose from 51% in control group to 84% in the cells grown with astro-CM containing 453 pg secreted GDNF, an increase that was highly significant (P < 0.0001). However, larger volumes of the GDNF-enriched conditioned medium failed to improve cell survival and addition of volumes that contained 1,600 pg or more GDNF further reduced survival rate to below 70%. Changes in cell survival paralleled to changes in the percent of apoptotic cell morphologies. These data demonstrate the feasibility of using astrocytes as minipumps to stably oversecrete neurotrophic factors and further indicate that GDNF can be applied to neuroprotection studies in PD pending the optimization of its concentrations.

Keywords

GDNF Dopaminergic Astrocytes Neuroprotection Lentivirus Lentiviral vectors Parkinson's disease

Abbreviations

Astro-CM

Astrocytic conditioned medium

GDNF

Glial cell line-derived neurotrophic factor

6-OHDA

6-Hydroxy dopamine

TH

Tyrosine hydroxylase

SNpc

Substantia nigra pars compacta

SN-ST

Nigrostriatal

Copyright information

© Springer Science+Business Media, LLC 2011