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2 concentration were continuously monitored, and the latter three were maintained within the ranges of 140 to 180 beats per minute. 37.5° to 39.0°C, and 3.5 to 4.5%, respectively. Each animal was mounted in a stereotaxic frame, and in order to minimize pulsations arising from the heartbeat and respiration, the cisterna magna was cannulated, a bilateral pneumothorax was performed, and the animal was suspended from the stereotaxic frame. A craniotomy (3 to 4 mm) was made overlying the representation of the area centralis of area 17. After the surgery, the animals were paralyzed with pancuronium bromide (3 mg/kg), followed by a continuous infusion of 3 mg/kg per hour (i.V.). The eyes were focused on the screen of a computer monitor at a distance of 57 cm by means of an appropriate pair of gas-permeable contact lenses. The nictitating membranes were retracted and the pupils were dilated by local application of phenylephrine and atropine, respectively, A small opening was made in the dura, and a micropipette was positioned just above the cortical surface. A 4% mixture of agar in Ringer solution was applied to the cortical surface to reduce pulsations. Within 10 to 12 hours of recording and staining of the first cell, each animal was given a lethal injection of nembutal and perfused through the heart with phosphate-buffered saline (PBS), followed by PBS containing 2% paraformaldehyde and 1.25% glutaraldehyde. This protocol was approved by the University of California, Davis, and Yale University Institutional Animal Care and Use Committees and conforms to the guidelines recommended in Preparation and Maintenance of Higher Mammals During Neuroscience Experiments, NIH publication No. 91-3207 (National Institutes of Health, Bethesda, MD, 1991).
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+ -acetate and 2% biocytin, and beveled to a final resistance of 70 to 120 megohms. After each recording, the depth of the electrode tip was noted and the position of the recording site was marked on the skull. A new site was chosen at least 1 mm away from any previous recording site and the entire procedure was repeated. Visual stimuli were generated by a personal computer and displayed on a 19-inch color monitor (80-Hz noninterlaced refresh rate; 1024 × 768 resolution). The data were digitized at a rate of 20 kHz and processed offline. For each cell, we computed the peristimulus-time histogram (PSTH), the autocorrelation histogram (ACH), the power spectrum of the ACH, the interspike interval histogram (ISIH), and the power spectrum of the membrane potential fluctuations occurring spontaneously and in response to visual stimulation. For histology, a block of tissue containing the filled cells was sunk in 30% sucrose. Coronal sections 60 to 75 μm thick were made and were reacted for the presence of biocytin, with the use of standard techniques and diaminobenzidine visualization. The histological sections were cover-slipped and examined for the presence of labeled cells. Filled neurons were photographed and reconstructed with a camera lucida.
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Surprisingly, intrinsic firing patterns similar to those displayed by CH cells have rarely been observed in cortical in vitro slice preparations (8) [Y. Kang and F. Kayano, J. Neurophysiol. 72, 578 (1994)]. This raises the issue that cellular damage may account for the behavior of CH cells. We believe this interpretation to be highly unlikely for at least four reasons. First, using extracellular single-unit recordings in area 17 in both alert (7) and anesthetized (4, 6) cats and monkeys, we and others have observed a substantial number of cells having temporal firing characteristics virtually identical to those reported here for CH cells. Second, the consistency of the receptive field properties, morphology, and laminar positions of the CH cells indicate that they belong to a distinct category of cells. Third, we have observed CH cells in slices of the cat visual cortex in vitro, and the probability of finding these cells is greatly enhanced by the application of a muscarinic cholinergic agonist such as acetyl-β-methylcholine (26). This raises the possibility that neuromodulatory transmitters such as acetylcholine, which may be dramatically reduced in the slice, can modify the intrinsic properties of CH cells. This interpretation is also consistent with the recent observation that stimulation of the mesencephalic reticular formation enhances the occurrence and magnitude of synchronous cortical oscillations in the gamma frequency band (27). Fourth, injury, or the decay of recording quality, did not induce any of the "non-CH" cells to become CH neurons.
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We selected a sample of 28 visually responsive cells that we recorded long enough to collect at least 20 responses to the presentation of an optimally oriented drifting light bar. The spike trains were extracted from each trial and quantified with the use of two measures: the mean firing rate across trials for a 500-ms epoch centered on the peak of the visual response and the percentage of interspike intervals occurring during the visual responses that were <3 ms in duration. For the subthreshold membrane potential, we visually selected two 512-ms epochs from each trial, one during the period of spontaneous activity and the other centered on the peak of the visual response. For both epochs on each trial, we applied a seven-point median filter to remove the action potentials and stored the resulting signals at a resolution of 1 ms. The data were digitally filtered (10 to 100 Hz) to remove both low- and high-frequency components. The power spectrum was then computed for each epoch of data. The sum of all values in each spectrum between 25 and 65 Hz was computed and the distributions were compared, after normalization to the largest value in the data set, with the use of the Kolmogorov-Smirnov test.
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The firing rates (in spikes per second; means ± SD) of the four cell types were as follows: RS (n = 10), 26 ± 7; IB (n = 5), 18 ± 1; FS (n = 2), 27 ± 1; and CH (n = 11), 45.4 ± 22. RS > IB (P < 0.03); CH > RS (P < 0.009); and CH > IB (P < 0.002).
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The percentages of intervals <3 ms for the four cell types (means ± SD) were as follows: RS (n = 10), 0.6 ± 1.2; IB (n = 5), 1.4 ± 2.1; FS (n = 2), 3.8 ± 4.6; and CH (n = 11), 27.5 ± 10.8. CH > RS (P < 0.0003); CH > IB (P < 0.007).
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Synchronous gamma-band activity is prevalent in the hippocampus of behaving rats [A. Bragin et al., J. Neurosci. 15, 47 (1995)] and can also be recorded in the hippocampal and neocortical slice preparations [M. A. Whittington, R. D. Traub, J. G. R. Jefferys, Nature 373, 612 (1995)], In the hippocampal slice, gamma-band activity appears to be generated by networks of inhibitory interneurons [R. D. Traub, M. A. Whittington, S. B. Colling, G. Buzsaki, J. G. R. Jefferys, J Physiol. 493, 471 (1996)]. These data suggest that the cellular mechanisms generating gamma-band activity may be different for the hippocampus and visual cortex.
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M. Steriade, F. Amzica, D. Contreras, J. Neurosci. 16, 392 (1996); M. H. J. Munk, P. R. Roelfsema, P. König, A. K. Engel, W. Singer, Science 272, 271 (1996).
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M. Steriade, F. Amzica, D. Contreras, J. Neurosci. 16, 392 (1996); M. H. J. Munk, P. R. Roelfsema, P. König, A. K. Engel, W. Singer, Science 272, 271 (1996).
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Munk, M.H.J.1
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Singer, W.5
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note
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We thank L. Nowak and R. Azouz for their valuable contributions to this work. Supported by a grant from the National Eye Institute and a contract from the Office of Naval Research to C.M.G., and by grants from NIH and NSF to D.M.
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