Effects of striatal transplantation of cells transfected with GDNF gene without pre- and pro-regions in mouse model of Parkinson's disease.
Previously, we have shown that transgenic cells bearing the GDNF gene with deleted pre- and pro-regions (mGDNF) can release transgenic GDNF. The medium conditioned by transgenic cells with mGDNF induced axonal growth in rat embryonic spinal ganglion in vitro. Here we demonstrate a neurotrophic effect of mGDNF on 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.



HEK293 cells were transfected with a vector encoding an isoform of the human GDNF gene with deleted pre- and pro-regions (mGDNF). This factor in the medium conditioned by the transfected cells was shown to induce axonal growth in PC12 cells. The early Parkinson's disease model was established by injection of the dopaminergic pro-neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) into C57Bl/6 mice. Transgenic HEK293/mGDNF/GFP cells were transplanted into the striatum (caudate-putamen) of experimental mice. The sleep-wakefulness cycle was studied by continuous EEG and motor activity monitoring 1 and 2 weeks after MPTP injection. After the experiment, the motor coordination of experimental animals was evaluated in the rotarod test, and dopaminergic neurons in the substantia nigra pars compacta were counted in cross-sections of the midbrain. MPTP administration lowered the number of tyrosine hydroxylase immunopositive cells in the substantia nigra pars compacta, decreased motor coordination, and increased the total wake time during the dark period. The transplantation of HEK293/mGDNF cells into the caudate-putamen 3 days prior to MPTP injection smoothed these effects, while the control transplantation of HEK293 cells showed no notable impact.



Transplantation of transgenic cells with the GDNF gene lacking the pre- and pro-sequences can protect dopaminergic neurons in the mouse midbrain from the subsequent administration of the pro-neurotoxin MPTP, which is confirmed by polysomnographic, behavioral and histochemical data. Hence it is released from transfected cells and preserves the differentiation activity and neuroprotective properties.
Laboratory of Neurogenetic and Developmental Genetic, Institute of Gene Biology, Russian Academy of Sciences, Vavilova Str., 34/5, Moscow, Russia, 119334.|Ltd Apto-pharm, Kolomensky Road, 13A, Moscow, Russia, 115446.
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Revishchin
Department of Higher Nervous Activity, Faculty of Biology, M.V. Lomonosov Moscow State University, Lenin Hills d. 1, pp. 12, Moscow, Russia, 119234.|A.N. Severtsov Institute of Ecology and Evolution, Russian Academy of Sciences, Moscow, Russia.
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Moiseenko
Laboratory of Neurogenetic and Developmental Genetic, Institute of Gene Biology, Russian Academy of Sciences, Vavilova Str., 34/5, Moscow, Russia, 119334.|Ltd Apto-pharm, Kolomensky Road, 13A, Moscow, Russia, 115446.
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Kust
Department of Higher Nervous Activity, Faculty of Biology, M.V. Lomonosov Moscow State University, Lenin Hills d. 1, pp. 12, Moscow, Russia, 119234.|Research Institute of General Pathology and Pathophysiology, Russian Academy of Sciences, Moscow, Russia, 125315.
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Bazhenova
Laboratory of Neurogenetic and Developmental Genetic, Institute of Gene Biology, Russian Academy of Sciences, Vavilova Str., 34/5, Moscow, Russia, 119334.
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Teslia
Laboratory of Neurogenetic and Developmental Genetic, Institute of Gene Biology, Russian Academy of Sciences, Vavilova Str., 34/5, Moscow, Russia, 119334.
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Panteleev
A.N. Severtsov Institute of Ecology and Evolution, Russian Academy of Sciences, 33 Leninskij Prosp., Moscow, Russia, 119071.
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Kovalzon
Laboratory of Neurogenetic and Developmental Genetic, Institute of Gene Biology, Russian Academy of Sciences, Vavilova Str., 34/5, Moscow, Russia, 119334. lkorochkin@mail.ru.|Ltd Apto-pharm, Kolomensky Road, 13A, Moscow, Russia, 115446. lkorochkin@mail.ru.
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Pavlova
D000818:Animals; D017690:Cell Transplantation; D017077:Culture Media, Conditioned; D059290:Dopaminergic Neurons / Q000473:pathology / Q000502:physiology; D015316:Genetic Therapy; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008810:Mice, Inbred C57BL; D009043:Motor Activity / Q000502:physiology; D017072:Neostriatum / Q000378:metabolism / Q000473:pathology; D000071446:Neuronal Outgrowth / Q000502:physiology; D016716:PC12 Cells; D020734:Parkinsonian Disorders / Q000473:pathology / Q000503:physiopathology* / Q000628:therapy*; D065842:Pars Compacta / Q000473:pathology / Q000503:physiopathology*; D051381:Rats; D012890:Sleep / Q000502:physiology; D014162:Transfection; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism; D014851:Wakefulness / Q000502:physiology
D017077:Culture Media, Conditioned; C495039:GDNF protein, human; D051100:Glial Cell Line-Derived Neurotrophic Factor; D049452:Green Fluorescent Proteins; D014446:Tyrosine 3-Monooxygenase
GDNF; Neurotrophic factor; Parkinson’s disease; Sleep-wakefulness cycle; Substantia nigra