An altered gravitational environment represents a unique challenge for biological systems that have evolved against gravitational background. Ground-based and space research indicates that the developing nervous system is potentially affected by exposure to hyper/microgravity. With the construction of the orbiting International Space Station long-term research on the nervous system will be possible. With this perspective, we started ground-based studies to characterize mouse behavioral responses to rotation-induced 2 g hypergravity, using a custom-made centrifuge device. Brain levels of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) as well as NGF and BDNF expression and mast cell distribution in heart and lung, were evaluated and correlated with the changes in mouse behavior upon hypergravity exposure. Hypergravity strongly affected the spontaneous activity of the animals, selectively modifying mouse behavioral repertoire. Such changes were mainly related to variations in brain levels of NGF, while BDNF was slightly affected, thus confirming a role for these neurotrophins in neuronal plasticity underlying experience-induced neurobehavioral changes. Moreover, gender differences were observed in both behavioral and neurobiological responses to hypergravity. These results indicate that changes in the gravitational environment might represent a useful tool to investigate the neurobiological and behavioral responses to stressors and may provide insights into the mechanisms underlying development and plasticity of nervous system in brain, heart, and lung.