Temporal lobe epilepsy (TLE) is the most common form of focal epilepsy. Though available anticonvulsants achieve a full seizure control for two-thirds of the TLE patients, one third does not respond sufficiently to these drugs and resection of the seizure area is the best possible treatment for them. One way to improve this is to identify key regulatory ion channels in excitatory synaptic function. Here, we investigated the ultrastructural localization of I) P/Q-type voltage-gated calcium channels (Cav2.1) at the presynaptic site triggering vesicle release and II) AMPA receptors mediating the post-synaptic response in glutamatergic synapses of the human temporal cortex by applying a modified freeze-fracture replica immunolabeling technique. Surgical resections were performed in strict accordance with institutional and European guidelines, and were provided by the Medical University Vienna (these authors), and the Clinics of Neurosurgery at Kepler-University Hospital at JKU in Linz (Prof. Andreas Gruber). For Cav2.1 channels, we found a non-homogenous distribution forming distinct clusters in the presynaptic active zone of dendritic spine and shaft synapses. The density of channels and size of clusters significantly increased in spine but not shaft synapses in TLE patients compared to non-epileptic controls. In contrast, AMPA receptors distributed evenly in the post-synaptic site, and with their numbers significantly upregulated in the dendritic spine and shaft synapses of TLE patients. These findings indicate an overall increase in the postsynaptic excitation and a more distinct regulation of presynaptic release specific for excitatory cortical synapses in TLE.