Galina Pavlova, Shamadykova DZ., Kust N., Panteleev D., Rybalkina E., Savchenko E., Kovalzon V. and Revishchin A.
Department of Neurobiology, Institute of Gene Biologe, Apto-Pharm, Russia
GDNF therapy is effective for treating degeneration of dopaminergic neurons in Parkinson''s disease. It is possible that pre-(α)pro-GDNF is needed for conventional neuron survival, and pre-(β) pro-GDNF serves as SOS system during traumatic injury of neurons or neurodegenerative diseases. To study ''pro'' region function during fast transport and factor induction properties several versions of modified GDNF were made. Secretion of GDNF into medium has been shown. Then modified GDNF were introduced into HEK293 cells. Condition media after transgenic cell culturing was added into culture medium of rat embrional spinal ganglion explant. Deletion of ''pro'' region essentially increases GDNF effects as neural inductor. Study of culture of dissociated spinal ganglion and calculation of neural sprouts yielded the same results. Deletion of both pre- and pro-regions enhances trophic activity of GDNF (mGDNF). Spinal ganglia cultured in the presence of medium conditioned by cells transfected with mGDNF exhibited active growth of β-3-tubulin-positive axons by day 4. Then we demonstrate neurotrophic effect of mGDNF for PC12 cells in vitro as well as its neuroprotective effect on dopaminergic neurons in the substantia nigra pars compacta in vivo as indicated by improved motor coordination and sleep-wakefulness cycle in the MPTP mouse model of Parkinson's disease. Research is supported by Grant from RNF No. 14-15-00942.
Keywords: GDNF, ''pro'' region, neurodegenerative diseases, transgenic cells.