Estimating extracellular spike waveforms from CA1 pyramidal cells with multichannel electrodes.

Extracellular (EC) recordings of action potentials from the intact brain are embedded in background voltage fluctuations known as the "local field potential" (LFP). In order to use EC spike recordings for studying biophysical properties of neurons, the spike waveforms must be separated from the LFP. Linear low-pass and high-pass filters are usually insufficient to separate spike waveforms from LFP, because they have overlapping frequency bands. Broad-band recordings of LFP and spikes were obtained with a 16-channel laminar electrode array (silicone probe). We developed an algorithm whereby local LFP signals from spike-containing channel were modeled using locally weighted polynomial regression analysis of adjoining channels without spikes. The modeled LFP signal was subtracted from the recording to estimate the embedded spike waveforms. We tested the method both on defined spike waveforms added to LFP recordings, and on in vivo-recorded extracellular spikes from hippocampal CA1 pyramidal cells in anaesthetized mice. We show that the algorithm can correctly extract the spike waveforms embedded in the LFP. In contrast, traditional high-pass filters failed to recover correct spike shapes, albeit produceing smaller standard errors. We found that high-pass RC or 2-pole Butterworth filters with cut-off frequencies below 12.5 Hz, are required to retrieve waveforms comparable to our method. The method was also compared to spike-triggered averages of the broad-band signal, and yielded waveforms with smaller standard errors and less distortion before and after the spike.

Tracheotomy improves experiment success rate in mice during urethane anesthesia and stereotaxic surgery.

Urethane anesthesia is frequently used for acute experiments on small rodents in physiology and neuroscience. Severe respiratory distress is a common side-effect of urethane anesthesia in many strains of mice. Associated complications interfere with completion of experiments, and as a consequence more animals must be sacrificed. During experiments with stereotaxic brain surgery, we found that intubation by means of tracheotomy is an efficient way to maintain patent airways in these animals. Artificial ventilation of the animals is not required. In this paper we describe a simple, fast and reliable method for intubation of mice in experiments that involve a stereotaxic instrument. The method proved considerably easier to learn and apply than conventional intubation through the oral route. The incidence of breathing problems decreased from 77% in untreated mice to 9% in those that underwent tracheotomy. In addition, the success rate for our acute electrophysiological experiments increased from 24 to 77%. We conclude that tracheotomy reduces the number of sacrificed animals, and saves time and labor.

Microstructure of a spatial map in the entorhinal cortex.

The ability to find one's way depends on neural algorithms that integrate information about place, distance and direction, but the implementation of these operations in cortical microcircuits is poorly understood. Here we show that the dorsocaudal medial entorhinal cortex (dMEC) contains a directionally oriented, topographically organized neural map of the spatial environment. Its key unit is the 'grid cell', which is activated whenever the animal's position coincides with any vertex of a regular grid of equilateral triangles spanning the surface of the environment. Grids of neighbouring cells share a common orientation and spacing, but their vertex locations (their phases) differ. The spacing and size of individual fields increase from dorsal to ventral dMEC. The map is anchored to external landmarks, but persists in their absence, suggesting that grid cells may be part of a generalized, path-integration-based map of the spatial environment.

Spatial representation in the entorhinal cortex.

As the interface between hippocampus and neocortex, the entorhinal cortex is likely to play a pivotal role in memory. To determine how information is represented in this area, we measured spatial modulation of neural activity in layers of medial entorhinal cortex projecting to the hippocampus. Close to the postrhinal-entorhinal border, entorhinal neurons had stable and discrete multipeaked place fields, predicting the rat's location as accurately as place cells in the hippocampus. Precise positional modulation was not observed more ventromedially in the entorhinal cortex or upstream in the postrhinal cortex, suggesting that sensory input is transformed into durable allocentric spatial representations internally in the dorsocaudal medial entorhinal cortex.

Hippocampal neurons responding to first-time dislocation of a target object.

To examine how hippocampal neurons respond to a mismatch between retrieved and actual experience, we trained rats to find a hidden platform at a particular location in an annular watermaze and then moved the platform. Several cells that were silent at the new platform location before the move fired vigorously when the rat found the goal. The new activity was paralleled by reduced discharge in a subset of simultaneously recorded interneurons. The pattern of activity returned toward its original configuration as the rat learned the new location. The activation of specific hippocampal neurons following dislocation of a target object may be essential for synaptic plasticity and adaptive modification of the animal's representation of the environment.

Place cells and place recognition maintained by direct entorhinal-hippocampal circuitry.

Place cells in hippocampal area CA1 may receive positional information from the intrahippocampal associative network in area CA3 or directly from the entorhinal cortex. To determine whether direct entorhinal connections support spatial firing and spatial memory, we removed all input from areas CA3 to CA1, thus isolating the CA1 area. Pyramidal cells in the isolated CA1 area developed sharp and stable place fields. Rats with an isolated CA1 area showed normal acquisition of an associative hippocampal-dependent spatial recognition task. Spatial recall was impaired. These results suggest that the hippocampus contains two functionally separable memory circuits: The direct entorhinal-CA1 system is sufficient for recollection-based recognition memory, but recall depends on intact CA3-CA1 connectivity.