The Palm-Sized Cryoprobe System Based on Refrigerant Expansion and Boiling and Its Application to an Animal Model of Epilepsy.

GOAL: The purpose of this study is to propose the palm-sized cryoprobe system based on a new concept and to suggest that the freezing technique could be used for treatment of epilepsy. METHODS: We propose herein a cryoprobe system based on the boiling effect that uses a specific refrigerants with a boiling point higher than that of liquid nitrogen yet low enough to result in cell necrosis. To evaluate and verify the effectiveness of the proposed system, cooling characteristics are investigated in agar. In addition, the system is applied to a Wistar rat brain-model, in which the epileptic activities are induced in advance by a potent epileptogenic substance. RESULTS: The design concept yielded the following benefits: 1) the selected refrigerant promotes sealing in the tank; 2) the tank can be made as compact as possible, limited only by the volume required for the refrigerant; 3) because the tank and probe units can be separated by a nonconducting, flexible, and high-pressure tube, the tank unit can be manipulated without disturbing the probe tip with mechanical vibrations and electrical noise. Although the agar experiments, we verified that the proposed system can uniquely and reproducibly create an ice ball. Moreover, in the rat experiments in vivo, it was confirmed that penicillin G-induced epileptic activities disappeared on freezing with the proposed system. CONCLUSIONS: The palm-sized system has desired characteristics and can apply for an animal model of epilepsy. SIGNIFICANCE: Results of in vivo experiments suggest that cryosurgery may be an effective treatment for epilepsy.

Penicillin-induced epileptiform activity elevates focal brain temperature in anesthetized rats.

To elucidate a relationship between changes in focal brain temperature and severity of abnormal brain activity, epileptiform discharges and behavioral seizures were induced by Penicillin G in anesthetized rats, and focal brain-temperature was measured. Penicillin G was injected into the right primary sensorimotor cortex (400IU/µl). After the injection, epileptiform discharges induced a temperature increase gradually by 0.65±0.24°C. Moreover, when behavioral seizures were induced by reducing the anesthesia level, the temperature was raised by 0.26±0.22°C. These results suggest that elevation of the focal brain temperature is associated with the severity of epileptic activity.

Taste responsiveness of fungiform taste cells with action potentials.

It is known that a subset of taste cells generate action potentials in response to taste stimuli. However, responsiveness of these cells to particular tastants remains unknown. In the present study, by using a newly developed extracellular recording technique, we recorded action potentials from the basolateral membrane of single receptor cells in response to taste stimuli applied apically to taste buds isolated from mouse fungiform papillae. By this method, we examined taste-cell responses to stimuli representing the four basic taste qualities (NaCl, Na saccharin, HCl, and quinine-HCl). Of 72 cells responding to taste stimuli, 48 (67%) responded to one, 22 (30%) to two, and 2 (3%) to three of four taste stimuli. The entropy value presenting the breadth of responsiveness was 0.158 +/- 0.234 (mean +/- SD), which was close to that for the nerve fibers (0.183 +/- 0.262). In addition, the proportion of taste cells predominantly sensitive to each of the four taste stimuli, and the grouping of taste cells based on hierarchical cluster analysis, were comparable with those of chorda tympani (CT) fibers. The occurrence of each class of taste cells with different taste responsiveness to the four taste stimuli was not significantly different from that of CT fibers except for classes with broad taste responsiveness. These results suggest that information derived from taste cells generating action potentials may provide the major component of taste information that is transmitted to gustatory nerve fibers.

Ionic currents underlying rhythmic bursting of ventral mossy cells in the developing mouse dentate gyrus.

The electrophysiological properties of mossy cells were examined in developing mouse hippocampal slices using whole-cell patch-clamp techniques, with particular reference to the dorsoventral difference. Dorsal mossy cells exhibited a higher spontaneous excitatory postsynaptic potential (EPSP) frequency and larger maximal EPSP amplitude than ventral mossy cells. On the other hand, the blockade of synaptic inputs with glutamatergic and GABAergic antagonists disclosed a remarkable dorsoventral difference in the intrinsic activity: none (0/27) of the dorsal mossy cells showed intrinsic bursting, whereas the majority (35/47) of the ventral mossy cells exhibited intrinsic rhythmic bursting. To characterize the ionic currents underlying the rhythmic bursting of mossy cells, we used somatic voltage-clamp recordings in the subthreshold voltage range. Ventral bursting cells possessed both hyperpolarization-activated current (Ih) and persistent sodium current (INaP), whereas dorsal and ventral nonbursting cells possessed Ih but no INaP. Blockade of Ih with cesium did not affect the intrinsic bursting of ventral mossy cells. In contrast, the blockade of INaP with tetrodotoxin or phenytoin established a stable subthreshold membrane potential in ventral bursting cells. The current-voltage curve of ventral bursting cells showed a region of tetrodotoxin-sensitive negative slope conductance between -55 mV and a spike threshold ( approximately -45 mV). On the other hand, no subthreshold calcium conductances played a significant role in the intrinsic bursting of ventral mossy cells. These observations demonstrate the heterogeneous electrophysiological properties of hilar mossy cells, and suggest that the subthreshold INaP plays a major role in the intrinsic rhythmic bursting of ventral mossy cells.

Stochastic resonance in the hippocampal CA3-CA1 model: a possible memory recall mechanism.

Stochastic resonance (SR) in a hippocampal network model was investigated. The hippocampal model consists of two layers, CA3 and CA1. Pyramidal cells in CA3 are connected to pyramidal cells in CA1 through Schaffer collateral synapses. The CA3 network causes spontaneous irregular activity (broadband spectrum peaking at around 3 Hz), while the CA1 network does not. The activity of CA3 causes membrane potential fluctuations in CA1 pyramidal cells. The CA1 network also receives a subthreshold signal (2.5 or 50 Hz) through the perforant path (PP). The subthreshold PP signals can fire CA1 pyramidal cells in cooperation with the membrane potential fluctuations that work as noise. The firing of the CA1 network shows typical features of SR. When the frequency of the PP signal is in the gamma range (50 Hz), SR that takes place in the present model shows distinctive features. 50 Hz firing of CA1 pyramidal cells is modulated by the membrane potential fluctuations, resulting in bursts. Such burst firing in the CA1 network, which resembles the firing patterns observed in the real hippocampal CA1, improves performance of subthreshold signal detection in CA1. Moreover, memory embedded at Schaffer collateral synapses can be recalled by means of SR. When Schaffer collateral synapses in subregions of CA1 are augmented three-fold as a memory pattern. pyramidal cells in the subregions respond to the subthreshold PP signal due to SR, while pyramidal cells in the rest of CA1 do not fire.

Complexity of spatiotemporal activity of a neural network model which depends on the degree of synchronization.

Spatiotemporal activity of a hippocampal CA3 model and its dynamic features were investigated. The CA3 model consists of 256 pyramidal cells and 25 inhibitory interneurons. Each pyramidal cell is a single-compartment model which was reduced from the 19-compartment cable model of the CA3 pyramidal cell developed by [Traub et al. (1991)]. Each interneuron is a model which causes tonic responses to constant depolarizing currents. The hippocampal model spontaneously causes four kinds of rhythms, A-D, which depend on the degree of synchronization of neuronal activity. The rhythm A (about 2Hz) which occurs in a range of strong mutual excitation is spatially coherent, though epileptiform bursts of pyramidal cells propagate from one end of the network to the other in a short period of time. The rhythm B (about 3Hz) occurs in an intermediate range of the strength of mutual excitation; synchronization of bursts is incomplete and the spatiotemporal pattern is complex. When the mutual excitation is relatively weak, the rhythm C (about 6Hz) occurs. Burst propagation is not uniform in direction, and the spatiotemporal activity is irregular. The rhythm D (10-35Hz) occurs in a range of weak mutual excitation when the recurrent inhibition is relatively strong. In this parameter region, pyramidal cells do not cause bursting discharges but irregular beating discharges. The hippocampal model causes phase-lockings and irregular responses to periodic synaptic stimulation depending on its own rhythmic activity and stimulus parameters. Bursting discharges of pyramidal cells are well synchronized in phase-locked responses. Several irregular responses of the rhythms A and B are evidently chaotic; each one-dimensional strobomap of chaotic responses is a non-invertible function with an unstable fixed point. Attractors reconstructed from chaotic responses demonstrate the stretching and folding mechanism.