Glutamate is the main excitatory neurotransmitter in the brain where, due to the activity of specific vesicular glutamate transporters, it accumulates in synaptic vesicles. The vesicular glutamate transporter 1 is found in the majority of axon terminals that form asymmetrical (excitatory) synapses in the rat neocortex. However, since there is no information available regarding the distribution of vesicular glutamate transporter 1 in the human neocortex, we have used correlative light and electron microscopy to define its expression in this tissue. We found that the distribution of vesicular glutamate transporter 1-immunoreactivity is virtually identical to that found in the rat neocortex, both at the light and electron microscope levels. Therefore, we assessed whether vesicular glutamate transporter 1 immunostaining might be a useful tool to study the pathological alterations of glutamatergic transmission in the epileptic cerebral cortex. We analyzed the distribution of vesicular glutamate transporter 1 in the peritumoral neocortex of patients with epilepsy secondary to low-grade tumors. In these regions, we found alterations in the pattern of vesicular glutamate transporter 1-immunoreactivity that perfectly matched the neuronal loss and gliosis, as well as the decrease in the number of asymmetrical synapses identified by electron microscopy in this tissue. Thus, vesicular glutamate transporter 1 immunostaining appears to be a reliable and simple tool to study glutamatergic synapses in the normal and epileptic human cerebral cortex.