Erwin-Josef Speckmann

Spreading depression in human neocortical slices

Cortical spreading depression (CSD) occurrence has been suggested to be associated with seizures, migraine aura, head injury and brain ischemia-infarction. Only few studies identified CSD in human neocortical slices and no comprehensive study so far evaluated this phenomenon in human. Using the neocortical tissue excised for treatment of intractable epilepsy, we aimed to investigate CSD in human. CSD was induced by KCl injection and by modulating T-type Ca(2+) currents in incubated human neocortical tissues in an interphase mode. The DC-fluctuations were recorded by inserting microelectrodes into different cortical layers. Local injection of KCl triggered single CSD that propagated at 3.1+/-0.1 mm/min. Repetitive CSD also occurred spontaneously during long lasting application (5 h) of the T-type Ca(2+) channel blockers amiloride (50 microM) or NiCl(2) (10 microM) which was concomitant with a reversible extracellular potassium increase up to 50 mM. CSD could be blocked by the N-methyl-D-aspartate receptor antagonist 2-amino-5-phosphonovaleric acid in all cases. The results demonstrate that modulation of the Ca(2+) dynamics conditioned human neocortical slices and increased their susceptibility to generate CSD. Furthermore, these data indicate that glutamatergic pathway plays a role in CSD phenomenon in human.

Spreading convulsions, spreading depolarization and epileptogenesis in human cerebral cortex

Spreading depolarization of cells in cerebral grey matter is characterized by massive ion translocation, neuronal swelling and large changes in direct current-coupled voltage recording. The near-complete sustained depolarization above the inactivation threshold for action potential generating channels initiates spreading depression of brain activity. In contrast, epileptic seizures show modest ion translocation and sustained depolarization below the inactivation threshold for action potential generating channels. Such modest sustained depolarization allows synchronous, highly frequent neuronal firing; ictal epileptic field potentials being its electrocorticographic and epileptic seizure its clinical correlate. Nevertheless, Leão in 1944 and Van Harreveld and Stamm in 1953 described in animals that silencing of brain activity induced by spreading depolarization changed during minimal electrical stimulations. Eventually, epileptic field potentials were recorded during the period that had originally seen spreading depression of activity. Such spreading convulsions are characterized by epileptic field potentials on the final shoulder of the large slow potential change of spreading depolarization. We here report on such spreading convulsions in monopolar subdural recordings in 2 of 25 consecutive aneurismal subarachnoid haemorrhage patients in vivo and neocortical slices from 12 patients with intractable temporal lobe epilepsy in vitro. The in vitro results suggest that γ-aminobutyric acid-mediated inhibition protects from spreading convulsions. Moreover, we describe arterial pulse artefacts mimicking epileptic field potentials in three patients with subarachnoid haemorrhage that ride on the slow potential peak. Twenty-one of the 25 subarachnoid haemorrhage patients (84%) had 656 spreading depolarizations in contrast to only three patients (12%) with 55 ictal epileptic events isolated from spreading depolarizations. Spreading depolarization frequency and depression periods per 24 h recording episodes showed an early and a delayed peak on Day 7. Patients surviving subarachnoid haemorrhage with poor outcome at 6 months showed significantly higher total and peak numbers of spreading depolarizations and significantly longer total and peak depression periods during the electrocorticographic monitoring than patients with good outcome. In a semi-structured telephone interview 3 years after the initial haemorrhage, 44% of the subarachnoid haemorrhage survivors had developed late post-haemorrhagic seizures requiring anti-convulsant medication. In those patients, peak spreading depolarization number had been significantly higher [15.1 (11.4–30.8) versus 7.0 (0.8–11.2) events per day, P = 0.045]. In summary, monopolar recordings here provided unequivocal evidence of spreading convulsions in patients. Hence, practically all major pathological cortical network events in animals have now been observed in people. Early spreading depolarizations may indicate a risk for late post-haemorrhagic seizures.

Comparison of Brain Extracellular Fluid, Brain Tissue, Cerebrospinal Fluid, and Serum Concentrations of Antiepileptic Drugs Measured Intraoperatively in Patients with Intractable Epilepsy

Summary:  Purpose: The mechanisms of drug resistance in epilepsy are only incompletely understood. According to a current concept, overexpression of drug efflux transporters at the blood–brain barrier may reduce levels of antiepileptic drugs (AEDs) in epileptogenic brain tissue. Increased expression of drug efflux transporters such as P‐glycoprotein has been found in brain tissue surgically resected from patients with medically intractable epilepsy, but it is not known whether this leads to decreased extracellular (interstitial) AED concentrations in affected brain regions. This prompted us to measure concentrations of AEDs in the extracellular space of human neocortical tissue by using intraoperative microdialysis (IOMD) in those parts of the brain that had to be removed for therapeutic reasons. For comparison, AED levels were determined in brain tissue, subarachnoid CSF, and serum.

Reduction of voltage-operated potassium currents by levetiracetam : a novel antiepileptic mechanism of action?

Levetiracetam (ucb L059; Keppra) is a novel antiepileptic drug. Its effects on action potential generation and voltage-operated potassium currents were studied in acutely isolated hippocampal CA1 neurones from rat and guinea pig, using the patch-clamp technique in the whole-cell configuration. (i) Levetiracetam reduced repetitive action potential generation and affected the single action potential. Levetiracetam, 100 microM, decreased the total number of action potentials and reduced the total depolarisation area of repetitive action potentials by 21%. Furthermore, levetiracetam increased the duration of the first action potential slightly, prolonged that of the second action potential by 13% and decreased the slope of rise by 23%. (ii) Levetiracetam decreased the voltage-operated potassium current. Without effect on sodium and A-type potassium currents, levetiracetam, 100 microM, reduced the delayed rectifier current by 26%. The concentration of half-maximal block was 47 microM for guinea pig and 6 microM for rat neurones. Thus, the reduction of repetitive action potential generation by levetiracetam can be attributed, unexpectedly, to a moderate reduction of the delayed rectifier potassium current, as supported by a simulation of action potential generation. This suggests that a reduction of potassium currents may contribute to the antiepileptic effect(s) of levetiracetam.

Electrogenesis of Cortical DC Potentials

Publisher Summary This chapter discusses electrogenesis of cortical DC potentials. In accordance with earlier suggestions the term “DC potentials” is used to characterize the whole frequency range of extracellular field potentials that are reliably recorded by means of DC registration techniques. With this definition DC potentials comprise sustained shifts and slow fluctuations as well as faster waves superimposed on deviations of the baseline. The various forms of event-related potentials that are dealt with in subsequent presentations and discussions can therefore be classified as special subtypes of DC potentials. The question is where and how DC field potentials recorded, for instance, from the scalp are generated. Compared to an analysis of the conventional EEG, studies on generator structures and mechanisms of DC potentials meet with additional difficulties. A severe problem is that sustained or slowly changing DC potentials can originate, in principle, from a variety of generators. The role played by neurons as DC potential generators seems of particular interest.

Cortico-spinal connections in the rat. I. Monosynaptic and polysynaptic responses of cervical motoneurons to epicortical stimulation

SummaryIn rats the fast cortico-spinal pathway and its linkage to motoneurons of the distal portions of the forelimbs were examined by electrophysiological techniques:1.Unilateral epicortical stimulation with a single pulse led to “flick”-like movements of corresponding muscle groups in the ipsi- and contralateral limbs. The effective stimulation points of the forelimbs were localized in fronto-central areas, those of the hindlimbs in centroparietal regions of the cortex.2.After a single epicortical stimulus a cord dorsum potential appeared in all cervical, thoracal and lumbar segments, the early part of which could be shown to reflect the activity of fast cortico-spinal fibres conducting with a velocity of 60 m/s. The sites of origin of the cortical-spinal fibres mapped by antidromic stimulation exceeded the cortical regions determined as motor areas by orthodromic stimulation in parietal and occipital direction.3.Single stimuli applied to the ipsi- and contralateral cortical surface caused a sequence of EPSP in all studied cervical motoneurons of the distal forearm. In nearly half of the impaled neurons the first EPSP was monosynaptic. Postsynaptic reactions following ipsi- and contralateral epicortical stimulation did not show significant differences both in flexor and extensor motoneurons. A minimum increase of the stimulus intensity beyond threshold level of the first EPSP led to action potentials in the early part of the synaptic response. The results demonstrate that in non-primates, showing skilled movements of the hand and digits, a powerful direct cortico-motoneuronal connection exists which is of predominant functional significance.

Spreading Depression Enhances Human Neocortical Excitability in vitro

Cortical spreading depression (CSD) plays a role in migraine with aura. However, studies of the neuronal effects of CSD in human cortex are scarce. Therefore, in the present study, the effects of CSD on the field excitatory postsynaptic potentials (fEPSP) and the induction of long-term potentiation (LTP) were investigated in human neocortical slices obtained during epilepsy surgery. CSD significantly enhanced the amplitude of fEPSP following a transient suppressive period and increased the induction of LTP in the third layer of neocortical tissues. These results indicate that CSD facilitates synaptic excitability and efficacy in human neocortical tissues, which can be assumed to contribute to hyperexcitability of neocortical tissues in patients suffering from migraine.

Cerebral localization and regulation of the cell volume-sensitive serum- and glucocorticoid-dependent kinase SGK1

The serum- and glucocorticoid-dependent kinase SGK1 is regulated by alterations of cell volume, whereby cell shrinkage increases and cell swelling decreases the transcription, expression and activity of SGK1. The kinase is expressed in all human tissues studied including the brain. The present study was performed to localize the sites of SGK1 transcription in the brain, to elucidate the influence of the hydration status on SGK1 transcription and to explore the functional significance of altered SGK1 expression. Northern blot analysis of human brain showed SGK1 to be expressed in all cerebral structures examined: amygdala, caudate nucleus, corpus callosum, hippocampus, substantia nigra, subthalamic nucleus and thalamus. In situ hybridization and immunohistochemistry in the rat revealed increased expression of SGK1 in neurons of the hippocampal area CA3 after dehydration, compared with similar slices from brains of euvolaemic rats. Additionally, several oligodendrocytes, a few microglial cells, but no astrocytes, were positive for SGK1. The abundance of SGK1 mRNA in the temporal lobe, including hippocampus, was increased by dehydration and SGK1 transcription in neuroblastoma cells was stimulated by an increase of extracellular osmolarity. Co-expression studies in Xenopus laevis oocytes revealed that SGK1 markedly increased the activity of the neuronal K+ channel Kv1.3. As activation of K+ channels modifies excitation of neuronal cells, SGK1 may participate in the regulation of neuronal excitability.

Topography and bioelectrical properties of identifiable neurons in the buccal ganglion of Helix pomatia.

Abstract In the lateral part of the buccal ganglion of Helix pomatia three large neurons are found to exist. The present investigations aimed at identifying these cells both from the topographical and electrophysiological point of view. The results demonstrate that these neurons fulfill the requirements of a model structure. Because of their different activity patterns they can, moreover, be used for comparative studies.

Spreading depression enhances the spontaneous epileptiform activity in human neocortical tissues.

Spreading depression (SD) is a well‐known phenomenon in animal models of experimental epilepsy. However, the interaction of SD with epileptiform activity in human neuronal tissues is not clear. The aim of the present study was to investigate the effect of SD on synchronous rhythmic sharp field potentials in human neocortical slices. Spreading depression was elicited in human neocortical slices that exhibited sharp potentials. Extracellular field potentials were recorded from the third and fifth layers. SD significantly enhanced the repetition rate and amplitude of spontaneous rhythmic potentials in all tested slices. The results indicate that SD may facilitate the synchronization of different foci of rhythmic sharp field potentials and increase the excitability in human brain tissue.