Experiments have been performed on unanesthetized rats using a chronic restraint system. The animal's head was held in a stereotaxic apparatus by means of two metallic tubes fixed on the skull with dental cement. Electrodes consisted of a recording micropipette (filled with 1 M NaCl and 2% Pontamine Blue) attached to a multibarreled micropipette for iontophoresis. Electrode penetrations were reconstructed on camera lucida drawings of frontal brain sections. The overall percentage of spontaneously active somatosensory neurons was 77% with a mean spontaneous activity of 5.9 impulse/s (n = 405). Yet differences were observed in the proportions of active cells as well as in the mean spontaneous activity between cortical layers (both parameters being significantly higher in layers V and VI). Comparison with results obtained under urethane anesthesia [Dykes R. W. and Lamour Y. (1988) J. Neurophysiol. 60, 703-724] shows that the percentage of the spontaneously active neurons and the mean spontaneous activity were both significantly higher in unanesthetized rats (77 vs 36%; 5.9 vs 2.6 impulse/s). Nevertheless, the laminar distribution of the most active cells was similar under both conditions. In the present study, 52.3% of the neurons (n = 380) were excited by acetylcholine and 46% (n = 198) by carbachol. Significantly larger percentages of neurons excited by acetylcholine were found in layers Vb and VIb. These effects of cholinergic agonists--observed for the first time in unanesthetized rats--differed significantly from those previously obtained under anesthesia (33 and 34% of neurons excited by acetylcholine and carbachol, respectively) [Lamour Y. et al. (1982) Neuroscience 7, 1483-1494].(ABSTRACT TRUNCATED AT 250 WORDS)