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Availability of Pre- and Pro-regions of Transgenic GDNF Affects the Ability to Induce Axonal Sprout Growth

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Abstract

Plasmids containing four GFP-tagged isoforms of the human GDNF gene, with both pre- and pro-regions (pre–pro- GDNF), with the pre- (pre-GDNF) or the pro-region (pro-GDNF) alone, and without the pre- and pro-regions (mGDNF), were used to transfect HEK293 cells (human embryonic kidney cell line). The effect of the transgenic products on the growth of processes was studied in the spinal ganglia of 14-day rat embryos. Media conditioned by the transgenic cells were used to culture explants and dissociated cells of embryonic dorsal root ganglia attached to the bottom of the plate. Medium conditioned by gfp-transgenic HEK293 cells was used as the control. Spinal ganglia explants and dissociated cells cultured in a medium supplemented with recombinant GDNF (recGDNF) as well as in conditioned media containing the pre-GDNF and mGDNF products demonstrated active growth of processes immunopositive for neuronal marker beta-3-tubulin as early as on culture day 4. The ganglia and cells cultured in control medium and media conditioned by cells transgenic for pro-GDNF had no or very few processes even after 10 days of culture.

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Acknowledgments

This work was supported by State Contract no. 8150, Program “Molecular and Cell Biology” and grant Russian Science Foundation (14-15-00942). Experiments were performed using the equipment of the IGB RAS facilities supported by the Ministry of Science and Education of the Russian Federation (grants 16.552.11.7067).

Conflict of Interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Institute of Gene Biology, Russian Academy of Sciences, 34 Vavilov St., 119334, Moscow, Russia

    Nadezda Kust, Dmitri Panteleev, Ilya Mertsalov, Alexander Revishchin & Galina Pavlova

  2. Institute of Medicine and Transplantation of Cells, Moscow, Russia

    Ekaterina Savchenko, Alexander Revishchin & Galina Pavlova

  3. Ltd Apto-pharm, Moscow, Russia

    Nadezda Kust, Ekaterina Savchenko, Ekaterina Rybalkina, Alexander Revishchin & Galina Pavlova

  4. Department of Basic and Applied Neurobiology, V. P. Serbskii Center of Social and Forensic Psychiatry, Moscow, Russia

    Ekaterina Savchenko

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  1. Nadezda Kust
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Correspondence to Galina Pavlova.

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Kust, N., Panteleev, D., Mertsalov, I. et al. Availability of Pre- and Pro-regions of Transgenic GDNF Affects the Ability to Induce Axonal Sprout Growth. Mol Neurobiol 51, 1195–1205 (2015). https://doi.org/10.1007/s12035-014-8792-8

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  • DOI: https://doi.org/10.1007/s12035-014-8792-8

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