Pavlova G., Kust N., Panteleev D., Mertsalov I., Rybalkina E., Savchenko E., Revishchin A.

Institute of Gene Biology Russian Academy of Sciences, Ltd. Apto-pharm Moscow, Vavilov Str., 34/5 119334 Moscow Russia

Abstract:

GDNF is a major factor for a survival of the dopamine neurons of the midbrain. It supports the axon growth as well as survival of the neurons of that type. For the different models of the Parkinson disease it has been shown that GDNF could prevent the neurotoxically provoked death of the dopamine neurons and supports recuperation of its functional activity. Though some by-side effects are also known, like loosing of weight and possibility of neoplastic transformation. We prepared a genetic construct caring human GDNF, introduced it into HEK293 cells, and then transplanted the cells into parenchyma of the mouse brain. It has been shown that transgenic cells which express GDNF essentially reduce the glial scar formation. Therefore GDNF could be applied during transplantation into the brain to improve the transplant survival.  Though in humans GDNF gene supply two versions of mRNA: pre-(α)pro-GDNF and truncated pre-(β) pro-GDNF. Pre-(α)pro-GDNF is secreted through Golgi apparatus and pre-(β) pro-GDNF is located in the secretory vesicles and moves by fast secretion pathway. 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. A secretion of the factor into medium has been shown by western blot analysis. All modified GDNF were introduced into HEK293 cells, and transgenic cell lines were maintained. The condition media after culturing the cells with modified GDNF was added into culture medium of rat embrional spinal ganglion explant and a growth of neural sprouts were analyzed. Deletion of 'pro' region essentially increases of GDNF effects as neural inductor. A study of culture of dissociated spinal ganglion and calculation of neural sprouts yielded the same results.

Research is supported by Grants from RFBR, Ministry of Science and Education.