The aim of the investigation was to assess antihypoxic and neuroprotective properties of the glial cell line-derived neurotrophic factor (GDNF) in hypoxia models in vitro and in vivo. In vitro experiments were carried out on primary hippocampal cultures. Hypoxia modeling was performed on day 14 of culture development in vitro (DIV) by replacing the normoxic cultural medium with a medium containing low oxygen for 10 minutes. Registration of extracellular action potentials was conducted by MEA systems (Multichannel Systems, Germany) application. Study the effect of GDNF on synaptic plasticity was performed using SmartFlare RNA Detection Probes (Merck Millipore, France) and fluorescent microscopy. In vivo experiments were carried out on C57BL/6j male mice. For acute hypobaric hypoxia a vacuum flow-through chamber was used at the ambient temperature of 20–22°C. We have investigated the resistance of animals to hypoxia and their spatial memory retention in Morris water maze test 24 hours after hypoxia. In vitro and in vivo data demonstrated that GDNF has strong antihypoxic and neuroprotective properties. Preventive GDNF application before hypoxia contributed to the animal survival and spatial memory retention as well as the maintenance of cells viability in primary hippocampal cultures.