ABSTRACT
OBJECTIVE: Large-scale synchronous firing of neurons during seizures is modulated by electrotonic coupling between neurons via gap junctions. To explore roles for connexin36 (Cx36) gap junctions in seizures, we examined the seizure threshold of connexin36 knockout (Cx36KO) mice using a pentylenetetrazol (PTZ) model. METHODS: Mice (2-3months old) with Cx36 wildtype (WT) or Cx36KO genotype were treated with vehicle or 10-40mg/kg of the convulsant PTZ by intraperitoneal injection. Seizure and seizure-like behaviors were scored by examination of video collected for 20min. Quantitative real-time PCR (QPCR) was performed to measure potential compensatory neuronal connexin (Cx30.2, Cx37, Cx43 and Cx45), pannexin (PANX1 and PANX2) and gamma-aminobutyric acid type A (GABA(A)) receptor α1 subunit gene expression. RESULTS: Cx36KO animals exhibited considerably more severe seizures; 40mg/kg of PTZ caused severe generalized (≥grade III) seizures in 78% of KO, but just 5% of WT mice. A lower dose of PTZ (20mg/kg) induced grade II seizure-like behaviors in 40% KO vs. 0% of WT animals. There was no significant difference in either connexin, pannexin or GABA(A) α1 gene expression between WT and KO animals. CONCLUSION: Increased sensitivity of Cx36KO animals to PTZ-induced seizure suggests that Cx36 gap junctional communication functions as a physiological anti-convulsant mechanism, and identifies the Cx36 gap junction as a potential therapeutic target in epilepsy.
Subject(s)
Behavior, Animal/drug effects , Connexins/physiology , Seizures/chemically induced , Seizures/psychology , Animals , Connexins/genetics , Connexins/metabolism , Convulsants , DNA, Complementary/biosynthesis , DNA, Complementary/genetics , Female , Gap Junctions/metabolism , Injections, Intraperitoneal , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Nerve Tissue Proteins/metabolism , Neural Pathways/physiology , Pentylenetetrazole , Receptors, GABA-A/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Synapses/physiology , Gap Junction delta-2 ProteinABSTRACT
Neural activity can be modulated by applying a polarizing low-frequency (<<100 Hz) electric field (PLEF). Unlike conventional pulsed stimulation, PLEF stimulation has a graded, modulatory effect on neuronal excitability, and permits the simultaneous recording of neuronal activity during stimulation suitable for continuous feedback control. We tested a prototype system that allows for simultaneous PLEF stimulation with minimal recording artifact in a chronic tetanus toxin animal model (rat) of hippocampal epilepsy with spontaneous seizures. Depth electrode local field potentials recorded during seizures revealed a characteristic pattern of field postsynaptic potentials (fPSPs). Sinusoidal voltage-controlled PLEF stimulation (0.5-25 Hz) was applied in open-loop cycles radially across the CA3 of ventral hippocampus. For stimulated seizures, fPSPs were transiently entrained with the PLEF waveform. Statistical significance of entrainment was assessed with Thomson's harmonic F-test, with 45/132 stimulated seizures in four animals individually demonstrating significant entrainment (p < 0.04). Significant entrainment for multiple presentations at the same frequency (p < 0.01) was observed in three of four animals in 42/64 stimulated seizures. This is the first demonstration in chronically implanted freely behaving animals of PLEF modulation of neural activity with simultaneous recording.
Subject(s)
Electromagnetic Fields , Epilepsy/physiopathology , Hippocampus/physiopathology , Hippocampus/radiation effects , Seizures/physiopathology , Animals , Artifacts , Disease Models, Animal , Electric Stimulation/methods , Electrodes, Implanted , Electroencephalography , Epilepsy/chemically induced , Male , Rats , Rats, Sprague-Dawley , Seizures/chemically induced , Signal Processing, Computer-Assisted , Synaptic Potentials , Tetanus ToxinABSTRACT
State of vigilance is determined by behavioral observations and electrophysiological activity. Here, we improve automatic state of vigilance discrimination by combining head acceleration with EEG measures. We incorporated biaxial dc-sensitive microelectromechanical system (MEMS) accelerometers into head-mounted preamplifiers in rodents. Epochs (15s) of behavioral video and EEG data formed training sets for the following states: Slow Wave Sleep, Rapid Eye Movement Sleep, Quiet Wakefulness, Feeding or Grooming, and Exploration. Multivariate linear discriminant analysis of EEG features with and without accelerometer features was used to classify behavioral state. A broad selection of EEG feature sets based on recent literature on state discrimination in rodents was tested. In all cases, inclusion of head acceleration significantly improved the discriminative capability. Our approach offers a novel methodology for determining the behavioral context of EEG in real time, and has potential application in automatic sleep-wake staging and in neural prosthetic applications for movement disorders and epileptic seizures.
