Components of neuronal chloride transport in rat and human neocortex.

Considerable evidence indicates disturbances in the ionic gradient of GABAA receptor-mediated inhibition of neurones in human epileptogenic tissues. Two contending mechanisms have been proposed, reduced outward and increased inward Cl⁻ transporters. We investigated the properties of Cl⁻ transport in human and rat neocortical neurones (layer II/III) using intracellular recordings in slices of cortical tissue. We measured the alterations in reversal potential of the pharmacologically isolated inhibitory postsynaptic potential mediated by GABAA receptors (IPSPA) to estimate the ionic gradient and kinetics of Cl⁻ efflux after Cl⁻ injections before and during application of selected blockers of Cl⁻ routes (furosemide, bumetanide, 9-anthracene carboxylic acid and Cs+). Neurones from human epileptogenic cortex exhibited a fairly depolarized reversal potential of GABAA receptor-mediated inhibition (EIPSP-A) of -61.9 ± 8.3 mV. In about half of the neurones, the EIPSP-A averaged -55.2 ± 5.7 mV, in the other half, 68.6 ± 2.3 mV, similar to rat neurones (-68.9 ± 2.6 mV). After injections of Cl⁻, IPSPA recovered in human neurones with an average time constant (τ) of 19.0 ± 9.6 s (rat neurones: 7.2 ± 2.4 s). We calculated Cl⁻ extrusion rates (1/τ) via individual routes from the τ values obtained in different experimental conditions, revealing that, for example, the K+-coupled Cl⁻ transporter KCC2 comprises 45.3% of the total rate in rat neurones. In human neurones, the total rate of Cl⁻ extrusion was 63.9% smaller, and rates via KCC2, the Na+-K+-2Cl⁻ transporter NKCC1 and the voltage-gatedCl− channelClCwere smaller than in rat neurones by 80.0%, 61.7% and 79.9%, respectively. The rate via anion exchangers conversely was 14.4% larger in human than in rat neurones. We propose that (i) KCC2 is the major route of Cl⁻ extrusion in cortical neurones, (ii) reduced KCC2 is the initial step of disturbed Cl⁻ regulation and (iii) reductions in KCC2 contribute to depolarizing EIPSP-A of neurones in human epileptogenic neocortex.

Irregular RNA splicing curtails postsynaptic gephyrin in the cornu ammonis of patients with epilepsy.

Anomalous hippocampal inhibition is involved in temporal lobe epilepsy, and reduced gephyrin immunoreactivity in the temporal lobe epilepsy hippocampus has been reported recently. However, the mechanisms responsible for curtailing postsynaptic gephyrin scaffolds are poorly understood. Here, we have investigated gephyrin expression in the hippocampus of patients with intractable temporal lobe epilepsy. Immunohistochemical and western blot analyses revealed irregular gephyrin expression in the cornu ammonis of patients with temporal lobe epilepsy and four abnormally spliced gephyrins lacking several exons in their G-domains were isolated. Identified temporal lobe epilepsy gephyrins have oligomerization deficits and they curtail hippocampal postsynaptic gephyrin and GABA(A) receptor α2 while interacting with regularly spliced gephyrins. We found that cellular stress (alkalosis and hyperthermia) induces exon skipping in gephyrin messenger RNA, which is responsible for curtailed postsynaptic gephyrin and GABA(A) receptor α2 scaffolds. Accordingly, we did not obtain evidence for gephyrin gene mutations in patients with temporal lobe epilepsy. Cellular stress such as alkalosis, for example arising from seizure activity, could thus facilitate the development of temporal lobe epilepsy by reducing GABA(A) receptor α2-mediated hippocampal synaptic transmission selectively in the cornu ammonis.

Imaging of a thoracic spinal nerve haemangioblastoma by three-dimensional digital angiography.

Haemangioblastomas occur sporadically or in association with Von Hippel-Lindau (VHL) disease. Occasionally, they are associated with intrathecal cauda equina nerve roots and rarely with spinal nerve roots. The occurrence of a completely extrathecal, thoracic spinal nerve haemangioblastoma is exceptional, with only one case previously described in the English-language literature. We report an extradural thoracic spinal nerve haemangioblastoma in a patient with VHL disease causing a progressive thoracic radiculopathy. The advantages of three-dimensional digital angiography will be discussed. Three-dimensional reconstruction of the images acquired by rotational digital subtraction angiography is one of the most promising developments to improve surgical planning for complex vascular lesions.

CSF flow imaging in the management of third ventriculostomy with a reversed fast imaging with steady-state precession sequence.

The aim of this study was evaluation of a fast and slow-flow sensitive 2D steady-state free-precession sequence for its capability to prove the patency of endoscopic third ventriculostomy (TVS) in obstructive hydrocephalus, and to exclude communicating third ventricle prior to TVS. We compared gated and ungated variants of this sequence for this purpose. Twenty-three patients with obstructive hydrocephalus underwent 36 MR examinations with a 2D reversed fast imaging with steady-state precession (PSIF) sequence in a retrospectively cardiac gated (cine) and a faster but ungated version beside T1- and T2-weighted sequences in three planes. Thirteen patients were examined both before and after TVS, 4 patients solely before, and 6 patients solely after TVS. Imaging diagnoses were compared with intraoperative findings and clinical findings after TVS. Preoperative diagnosis of non-communicating third ventricle and cisterns was intraoperatively confirmed in 16 of 17 cases. Preoperative MRI was inconclusive in 1 case. Postoperative MRI revealed sufficient TVS in 16 of 19 cases and obstructed TVS in 3 of 19 cases due to several reasons. Findings at MRI were consistent in 19 of 19 cases with the clinical course and intraoperative results. The faster but ungated PSIF sequence was found to be diagnostically equivalent to the cardiac gated cine sequence. The CSF flow imaging with a 2D reversed fast imaging with steady-state precession sequence in conjunction with conventional T1- and T2-weighted images is a fast and reliable tool for pre- and postoperative functional evaluation in third ventriculostomy.

A novel mechanism underlying drug resistance in chronic epilepsy.

The development of resistance to pharmacological treatment is common to many human diseases. In chronic epilepsy, many patients develop resistance to anticonvulsant drug treatment during the course of their disease, with the underlying mechanisms remaining unclear. We have studied cellular mechanisms underlying drug resistance in resected hippocampal tissue from patients with temporal lobe epilepsy by comparing two groups of patients, the first displaying a clinical response to the anticonvulsant carbamazepine and a second group with therapy-resistant seizures. Using patch-clamp recordings, we show that the mechanism of action of carbamazepine, use-dependent block of voltage-dependent Na(+) channels, is completely lost in carbamazepine-resistant patients. Likewise, seizure activity elicited in human hippocampal slices is insensitive to carbamazepine. In marked contrast, carbamazepine-induced use-dependent block of Na(+) channels and blocked seizure activity in vitro in patients clinically responsive to this drug. Consistent with these results in human patients, we also show that use-dependent block of Na(+) channels by carbamazepine is absent in chronic experimental epilepsy. Taken together, these data suggest that a loss of Na(+) channel drug sensitivity may constitute a novel mechanism underlying the development of drug-resistant epilepsy.