The state of neuronal microtubule polymerization is influenced by microtubule-associated proteins such as MAP2, which is specifically localized within neuronal dendrites and cell bodies. We have demonstrated that stimulation of spinal cord or cortical neurons in vitro with excitatory amino acids results in a dramatic modification of the neuronal cytoskeleton as monitored with antibodies against MAP2 and tubulin. Stimulation of cultures with glutamate receptor agonists induced a reorganization of MAP2 immunoreactivity into a distinctive network of bundles within certain neuronal cell bodies and their proximal neurites. The effect was not abolished by depolymerizing drugs such as nocodazole, or protein synthesis inhibitors. The effect was dependent upon the entry of sodium following depolarization and was not associated with neuronal damage. We suggest that in neurons the state of the neuronal cytoskeleton can be modulated by glutamate receptor activation acting through MAP2.