Under free field and closed-system stimulation conditions, we studied the frequency threshold curves, auditory spatial sensitivity and binaurality of neurons in the primary auditory cortex (AC) of the big brown bat, Eptesicus fuscus. All 298 recorded AC neurons discharged phasically. They were recorded at depths less than 1,000 microns with response latencies of 7-25 ms, best frequencies (BFs) of 28-97 kHz and minimum thresholds (MTs) of 8-90 dB SPL. They received excitatory inputs from the contralateral ear and either excitatory (EE) or inhibitory (EI) inputs from the ipsilateral ear. These cortical neurons were tonotopically organized along the anteroposterior axis of the AC. High best frequency neurons were located anteriorly and low best frequency neurons posteriorly. They were most sensitive to sounds delivered from a restricted region of the contralateral frontal auditory space (0 degree-50 degrees in azimuth and 2 degrees up, 15 degrees down in elevation). Frontal auditory space representation appears to be systematically arranged according to the tonotopic axis such that the lateral space is represented posteriorly and the middle space anteriorly. Cortical neurons sequentially isolated from an orthogonally penetrated electrode had similar frequency threshold curves, BFs, MTs, points of maximal auditory spatial sensitivity and binaurality. The EE and EI columns are organized concentrically such that the small number of centrally located EE columns were surrounded by an overwhelming number of EI columns. Using Nissl and Golgi stains as well as c-fos immunocytochemistry, we studied the cytoarchitecture, cell types and sound elicited Fos-like immunoreactivity in the primary AC of this bat species. The primary AC of this bat species can be described into molecular (137 microns), external granular (55 microns), external pyramidal (95 microns), internal granular (102 microns), internal pyramidal (191 microns) and multiform (120 microns) layers. The main type of cells distributed among these six layers are the small, medium and large pyramidal cells. Others include the stellate, horizontal, granular, fusiform, basket, and Martinotii cells. When stimulated with 30 kHz and 79 dB SPL sounds under natural conditions, bilaterally and symmetrically distributed Fos-like immunoreactive neurons were observed in about 20% of neurons in each AC. When stimulated under monaurally plugged conditions, 39-48% more of Fos-like immunoreactive neurons were observed in the ipsilateral AC. This finding supports the fact that the primary AC receives auditory inputs mainly from the contralateral ear.