Rena Orman

Autonomic consequences of kainic acid–induced limbic cortical seizures in rats: Peripheral autonomic nerve activity, acute cardiovascular changes, and death

Purpose: Autonomic consequences of seizures are common, but can be severe. We sought to define changes in autonomic activity from limbic cortical seizures and their impact on the heart.

Vagus nerve stimulation-induced bradyarrhythmias in rats

The autonomic consequences of seizures can be severe. Death can follow from autonomic overactivity that causes a parasympathetically mediated bradyarrhythmia. We studied the cardiovascular consequences of unilateral and bilateral stimulation of the distal segments of transected vagus nerve in rats anesthetized with urethane. The range of stimulation rates tested is comparable to the firing rates observed in vagus nerve during seizures. There was a consistent inverse relation between stimulus rate and heart rate with nodal block appearing at 5-10 Hz and minimum HR levels (cardiac standstill) occurring at 50 Hz. Cardiac standstill could last many seconds. Blood pressure during VNS was maintained during lower frequency VNS, but collapsed at frequencies > or =20 Hz to dramatically impair ventricular filling. Recovery of heart rate and blood pressure after VNS was rapid. In the presence of sympathetic co-activation (pharmacological or hypercapnia and/or hypoxia), mean arterial pressure was better maintained and there was much better ventricular filling, but cardiac performance was worse (e.g. ejection fraction derived from echocardiography). The combination of sympathetic and parasympathetic overactivity was sometimes associated with prolonged (> or =20 s) apneic periods during VNS. We conclude that an abrupt increase in parasympathetic activity on the order of 5 times the background of parasympathetic tone can produce transient bradyarrhythmias, and increases on the order of 20 times can produce cardiac standstill, sometimes accompanied by apnea. Our findings suggest that parasympathetically mediated bradyarrhythmia must be accompanied by airway obstruction to sustain parasympathetic overactivity and produce hypoxia to ultimately cause death.

Relation of autonomic and cardiac abnormalities to ventricular fibrillation in a rat model of epilepsy.

Cardiac autonomic, conduction, and structural changes may occur in epilepsy and may contribute to sudden unexpected death in epilepsy (SUDEP), e.g. by increasing the risk for ventricular fibrillation (VF). In a model of chronic seizures in rats, we sought to study (1) cardiac and autonomic derangements that accompany the epileptic state, (2) whether chronically seizing rats experienced more significant cardiac effects after severe acute seizures, and (3) the susceptibility of chronically seizing rats to VF arising from autonomic and hypoxemic changes, which commonly occur during seizures. Sprague-Dawely rats were injected with saline or kainic acid to induce chronic seizures. At 2-3 months or 7-11 months after injection, these rats were studied with both 12-lead electrocardiography (to assess heart rate variability and QT dispersion) and echocardiography under ketamine/xylazine or urethane anesthesia. Hearts were subsequently excised, weighed, and examined histologically. Epileptic rats exhibited decreased vagal tone, increased QT dispersion, and eccentric cardiac hypertrophy without significant cardiac fibrosis, especially at 7-11 months post-injection. Of these three findings, vagal tone was inversely correlated with heart weights. Epileptic rats exhibited diminished systolic function compared to controls after severe acute seizures. However, animals with long-standing chronic seizures were less susceptible to autonomic/hypoxemia-driven VF, and their susceptibility inversely correlated with mean left ventricular wall thickness on histology. On the basis of this model, we conclude that cardiac changes accompany epilepsy and these can lead to significant seizure-associated cardiac performance decreases, but these cardiac changes actually lower the probability of VF.

Hemispheric differences in protein kinase C βII levels in the rat amygdala: Baseline asymmetry and lateralized changes associated with cue and context in a classical fear conditioning paradigm

The amygdala is critically important for fear learning, and specific kinases have been implicated as contributors to the mechanisms that underlie learning. We examined levels of protein kinase C betaII (PKC betaII) in the left and right lateral and basolateral nuclei (LA/BLA) of the amygdala from animals that were classically fear conditioned with tones as cues and footshocks. Groups consisted of animals that received neither tones nor shocks, paired tones and shocks, or unpaired tones and shocks. At 1 h after conditioning, some animals from each group were used for biochemical measurements of PKC betaII levels and other animals were given probe trials to assess freezing behavior to cue and context. The levels of PKC betaII were greater in the left hemisphere in animals receiving neither tones nor shocks and animals receiving paired tones and shocks. PKC betaII levels were greater in the right hemisphere of animals receiving randomly presented tones and shocks. Freezing times to cue were long (>80% of probe trial time) in both the paired tone/shock and randomly unpaired tone/shock groups. Freezing times to context were long in the unpaired tone/shock group, but not the paired tone/shock group. Correlational analyses showed that freezing times to context, but not cue, precisely predicted the right/left relation of PKC betaII levels in the LA/BLA: the greater the time spent freezing to context, the greater the increase in right hemisphere PKC betaII levels. We conclude that fear conditioning causes hemisphere and input specific increases in PKC betaII in the rat LA/BLA.

Local axon collaterals of area CA1 support spread of epileptiform discharges within CA1, but propagation is unidirectional

CA3 and subiculum are hippocampal formation regions that can initiate seizure activity because each has a substantial intrinsic excitatory connectivity. We studied the intrinsic connectivity of area CA1 by exploring the spread of synchronous population discharges in ventral hippocampal slices from rats using a recording chamber that permitted multiple simultaneous extracellular recordings along all laminae of CA1. Brief single stimulus pulses were applied to stratum oriens (SO) or stratum radiatum (SR) on the CA3 side or the subicular side of CA1. In disinhibited slices, events triggered with SO or SR stimulation on the CA3‐side propagated over the proximo‐distal extent of CA1 with a maximal conduction velocity of 0.4 m/s, comparable with antidromic conduction velocities within CA1. By contrast, SO or SR stimuli applied on the subicular side of CA1 triggered events that did not spread “backward” toward CA3. These events are rapidly decremented in amplitude and duration. Whereas antidromic responses were largest when stimuli were applied on the subicular side of CA1, such responses were not sufficient to trigger epileptiform discharges when excitatory transmission was intact. We conclude that the unidirectional spread of epileptiform activity in area CA1 is the result of an intrinsic axon collateral system where each pyramidal cell has a proportionally larger projection toward subiculum. Although this collateral system is sparse compared with other hippocampal formation regions, its unidirectionality protects against re‐entrant activation of CA3 and may be physiologically significant as a relay from proximal CA1 to distal CA1.

Computer simulation of epilepsy: Implications for seizure spread and behavioral dysfunction

Hippocampal area CA3 has been one of the most intensively studied brain regions for computer models of epileptiform activity. As physiological studies begin to extend outward to other hippocampal and parahippocampal areas, we must extend these models to understand more complex circuitry containing diverse elements. Study of subiculum is of particular interest in this context, as it is a structure of intermediate complexity, with an inchoate columnar and laminar organization. In addition to helping us understand seizures, modeling of these structures will also help us understand the genesis of physiological activity patterns that are below threshold for seizure generation. Such modeling can also serve as a basis for speculation regarding the nonictal behavioral consequences of epilepsy.

Laryngospasm, central and obstructive apnea during seizures: Defining pathophysiology for sudden death in a rat model

Seizure spread into the autonomic nervous system can result in life-threatening cardiovascular and respiratory dysfunction. Here we report on a less-studied consequence of such autonomic derangements-the possibility of laryngospasm and upper-airway occlusion. We used parenteral kainic acid to induce recurring seizures in urethane-anesthetized Sprague Dawley rats. EEG recordings and combinations of cardiopulmonary monitoring, including video laryngoscopy, were performed during multi-unit recordings of recurrent laryngeal nerve (RLN) activity or head-out plethysmography with or without endotracheal intubation. Controlled occlusions of a tracheal tube were used to study the kinetics of cardiac and respiratory changes after sudden obstruction. Seizure activity caused significant firing increases in the RLN that were associated with abnormal, high-frequency movements of the vocal folds. Partial airway obstruction from laryngospasm was evident in plethysmograms and was prevented by intubation. Complete glottic closure (confirmed by laryngoscopy) occurred in a subset of non-intubated animals in association with the largest increases in RLN activity, and cessation of airflow was followed in all obstructed animals within tens of seconds by ST-segment elevation, bradycardia, and death. Periods of central apnea occurred in both intubated and non-intubated rats during seizures for periods up to 33s and were associated with modestly increased RLN activity, minimal cardiac derangements, and an open airway on laryngoscopy. In controlled complete airway occlusions, respiratory effort to inspire progressively increased, then ceased, usually in less than 1min. Respiratory arrest was associated with left ventricular dilatation and eventual asystole, an elevation of systemic blood pressure, and complete glottic closure. Severe laryngospasm contributed to the seizure- and hypoxemia-induced conditions that resulted in sudden death in our rat model, and we suggest that this mechanism could contribute to sudden death in epilepsy.

Efferent and afferent vagal actions on cortical blood flow and kainic acid-induced seizure activity in urethane anesthetized rats

Autonomic dysfunction during seizures can induce bradyarrhythmia via efferent vagal overactivity. We studied cardiovascular, brain blood flow, and electroencephalographic consequences of vagal stimulation during seizures in rats. Efferent vagal stimulation reduced seizure activity, completely suppressing it at high frequencies, by reducing heart rate, arterial pressure, and cortical blood flow. Afferent vagal activation was more variable, and the highest stimulation frequencies also appeared to reduce cortical blood flow. We conclude that efferent vagal activity can arrest ongoing seizure activity by ultimately decreasing hippocampal blood flow. Afferent vagal activity (which does not occur during seizures) may have a similar action.

Broadening of Activity with Flow across Neural Structures

Synfire chains have long been suggested as a substrate for perception and information processing in the nervous system. However, embedding activation chains in a densely connected nervous matrix risks spread of signal that will obscure or obliterate the message. We used computer modeling and physiological measurements in rat hippocampus to assess this problem of activity broadening. We simulated a series of neural modules with feedforward propagation and random connectivity within each module and from one module to the next. We found that activity broadened as it propagated from one module to the next. This occurred over a wide array of parameters with greater broadening seen with increasing excitatory – excitatory synaptic strength. Activity broadening correlated positively with propagation velocity. Multi-electrode measurements of activity propagation in disinhibited CA1 slice demonstrated broadening of about 50% over 1 mm. Such broadening is a problem for information transfer that must be dealt with in a fully functioning nervous system.

Obstructive apnea due to laryngospasm links ictal to postictal events in SUDEP cases and offers practical biomarkers for review of past cases and prevention of new ones

Seizure spread into autonomic and respiratory brainstem regions is thought to play an important role in sudden unexpected death in epilepsy (SUDEP). As the clinical dataset of cases of definite SUDEP available for study grows, evidence points to a sequence of events that includes postictal apnea, bradycardia, and asystole as critical events that can lead to death. One possible link between the precipitating seizure and the critical postictal sequence is seizure‐driven laryngospasm sufficient to completely obstruct the airway for an extended period, but ictal laryngospasm is difficult to fully assess. Herein, we demonstrate in a rat model how the electrical artifacts of attempts to inspire during airway obstruction and features of the cardiac rhythm establish this link between ictal and postictal activity and can be used as practical biomarkers of obstructive apnea due to laryngospasm or other causes of airway obstruction.