Growing evidence shows that physical microenvironments and mechanical stresses, independent of soluble factors, help influence mesenchymal-stem-cell fate. rMSCs (rat mesenchymal stem cells) present spread, spindle shape when cultured in normal gravity (NG) while in simulated microgravity (SMG) they become unspread, round shape. Here we demonstrate that simulated microgravity can enhance the differentiation of mesenchymal stem cells into neurons, which might be a new strategy for the treatment of central nervous system diseases. rMSCs were cultured respectively in normal gravity and in a clinostat to simulate microgravity, followed with neuronal differentiated medium. The neuronal cells derived from rMSCs in SMG express higher microtubule-associated protein-2 (MAP-2), tyrosine hydroxylase (TH) and choline acetyltransferase (CHAT). Furthermore, as rMSCs are subjected to SMG, they excrete more neurotrophins like nerve growth factor (NGF), brain derived neurophic factor (BDNF) and ciliary neurotrophic factor (CNTF). Neuronal cells from SMG group generated more mature action potentials and displayed repetitive action potentials by comparison to cells from NG group. We conclude that simulated microgravity can enhance the differentiation of mesenchymal stem cells into neurons.