[Show abstract][Hide abstract] ABSTRACT: Introduction:
Familial hemiplegic migraine (FHM) is a rare autosomal dominant subtype of migraine with aura. The FHM3 subtype is caused by mutations in SCN1A, which is also the most frequent epilepsy gene encoding the voltage-gated Na(+) channel NaV1.1. The aim of this study was to explore the clinical, genetic and pathogenetic features of a pure FHM3 family.
A three-generation family was enrolled in this study for genetic testing and assessment of clinical features. Whole cell patch-clamp was performed to determine the functions of identified mutant NaV1.1 channels, which were transiently expressed in human tsA201 cells together with β1 and β2 subunits.
Results and conclusions:
We identified a novel SCN1A (p.Leu1624Pro) mutation in a pure FHM family with notably early-onset attacks at mean age of 7. L1624P locates in S3 of domain IV, the same domain as two of four known pure FHM3 mutations. Compared to WT channels, L1624P displayed an increased threshold-near persistent current in addition to other gain-of-function features such as: a slowing of fast inactivation, a positive shift in steady-state inactivation, a faster recovery and higher channel availability during repetitive stimulation. Similar to the known FHM3 mutations, this novel mutation predicts hyperexcitability of GABAergic inhibitory neurons.
[Show abstract][Hide abstract] ABSTRACT: Essential tremor (ET) is the most prevalent movement disorder, affecting millions of people in the United States. Although
a positive family history is one of the most important risk factors for ET, the genetic causes of ET remain unknown. In an
attempt to identify genetic causes for ET, we performed whole-exome sequencing analyses in a large Spanish family with ET,
in which two patients also developed epilepsy. To further assess pathogenicity, site-directed mutagenesis, mouse and human
brain expression analyses, and patch clamp techniques were performed. A disease-segregating mutation (p.Gly1537Ser) in the
SCN4A gene was identified. Posterior functional analyses demonstrated that more rapid kinetics at near-threshold potentials altered
ion selectivity and facilitated the conductance of both potassium and ammonium ions, which could contribute to tremor and
increase susceptibility to epilepsy, respectively. In this report, for the first time, we associated the genetic variability
of SCN4A with the development of essential tremor, which adds ET to the growing list of neurological channelopathies.
No preview · Article · Oct 2015 · Human Molecular Genetics
[Show abstract][Hide abstract] ABSTRACT: To examine rare KCNJ18 variations recently reported to cause sporadic and thyrotoxic hypokalaemic periodic paralysis (TPP).
We sequenced KCNJ18 in 474 controls (400 Caucasians, 74 male Asians) and 263 unrelated patients with periodic paralysis (PP), including 30 patients with TPP without mutations in established PP genes.
In 10 patients without TPP, we identified 9 heterozygous, novel variations (c.-3G>A, L15S, R81C, E273X, T309I, I340T, N365S, G394R, R401W) and a questionable heterozygous causative R399X stop variant. Studies on 40 relatives of these 10 patients showed that none of the variants were de novo in the patients and that R399X occurred in 3 non-affected relatives. Most affected amino acids lacked conservation and several clinically affected relatives did not carry the patient's variant. T309I, however, could be pathogenic under the pre-requisite of strongly reduced penetrance in females. Of the controls, 17 revealed 12 novel rare variants including the heterozygous E273X stop variant in three individuals.
Our study shows many different, rare KCNJ18 alterations in patients as well as controls. Only perhaps one meets the requirements of a disease-causing mutation. Therefore, KCNJ18 alterations are seldom pathogenic. Additional studies are required before patients with PP can be genetically diagnosed on the basis of a KCNJ18 variant alone.
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Full-text · Article · Apr 2015 · Journal of neurology, neurosurgery, and psychiatry
[Show abstract][Hide abstract] ABSTRACT: Febrile seizures affect 2–4% of all children and have a strong genetic component. Recurrent mutations in three main genes (SCN1A, SCN1B and GABRG2) have been identified that cause febrile seizures with or without epilepsy. Here we report the identification of mutations in STX1B, encoding syntaxin-1B, that are associated with both febrile seizures and epilepsy. Whole-exome sequencing in independent large pedigrees identified cosegregating STX1B mutations predicted to cause an early truncation or an in-frame insertion or deletion. Three additional nonsense or missense mutations and a de novo microdeletion encompassing STX1B were then identified in 449 familial or sporadic cases. Video and local field potential analyses of zebrafish larvae with antisense knockdown of stx1b showed seizure-like behavior and epileptiform discharges that were highly sensitive to increased temperature. Wild-type human syntaxin-1B but not a mutated protein rescued the effects of stx1b knockdown in zebrafish. Our results thus implicate STX1B and the presynaptic release machinery in fever-associated epilepsy syndromes.
[Show abstract][Hide abstract] ABSTRACT: Heterologous expression of sodium channel mutations in hypokalemic periodic paralysis reveals 2 variants on channel dysfunction. Charge-reducing mutations of voltage sensing S4 arginine residues alter channel gating as typically studied with expression in mammalian cells. These mutations also produce leak currents through the voltage sensor module, as typically studied with expression in Xenopus oocytes. DIIIS4 mutations at R3 in the skeletal muscle sodium channel produce gating defects and omega current consistent with the phenotype of reduced excitability. Here, we confirm DIIIS4 R3C gating defects in the oocyte expression system for fast inactivation and its recovery. We provide novel data for the effects of the cysteine mutation on voltage sensor movement, to further our understanding of sodium channel defects in hypokalemic periodic paralysis. Gating charge movement and its remobilization are selectively altered by the mutation at hyperpolarized membrane potential, as expected with reduced serum potassium.
[Show abstract][Hide abstract] ABSTRACT: For patients suffering from rare diseases it is often hard to find an expert clinician. Existing registries rely on manual registration procedures and cannot easily be kept up to date. A prototype data collection system for discovering experts on rare diseases using MEDLINE has been successfully deployed. Initial manual analyses demonstrate proof of concept and deliver promising results. Examining the associations between authors, diseases and MeSH-Terms is expected to open up a variety of possibilities beyond expert discovery.
Full-text · Article · May 2014 · Studies in health technology and informatics
[Show abstract][Hide abstract] ABSTRACT: We studied the consequences of the Nav1.4 mutation R1448H that is situated in the fourth voltage sensor of the channel and causes paramyotonia, a cold-induced myotonia followed by weakness. Previous work showed that the mutation uncouples inactivation from activation. We measured whole-cell Na(+) currents at 10, 15, 20, and 25°C using HEK293 cells stably transfected with wildtype (WT) and R1448H Na(+) channels. A Markov model was developed the parameters of which reproduced the data measured on WT and R1448H channels in the whole voltage and temperature range. It required an additional transient inactivated state and an additional closed-state inactivation transition not previously described. The model was used to predict single-channel properties, free energy barriers and temperature dependence of rates. It allowed us to draw the following conclusions: i) open-state inactivation results from a two-step process; ii) the channel re-openings that cause paramyotonia originate from enhanced deactivation/reactivation and not from destabilized inactivation; iii) the closed-state inactivation of R1448H is strikingly enhanced. We assume that latter explains the episodic weakness following cold-induced myotonia.
Full-text · Article · May 2014 · Acta myologica: myopathies and cardiomyopathies: official journal of the Mediterranean Society of Myology / edited by the Gaetano Conte Academy for the study of striated muscle diseases
[Show abstract][Hide abstract] ABSTRACT: Hypokalaemic periodic paralysis is typically associated with mutations of voltage sensor residues in calcium or sodium channels of skeletal muscle. To date, causative sodium channel mutations have been studied only for the two outermost arginine residues in S4 voltage sensor segments of domains I to III. These mutations produce depolarization of skeletal muscle fibres in response to reduced extracellular potassium, owing to an inward cation-selective gating pore current activated by hyperpolarization. Here, we describe mutations of the third arginine, R3, in the domain III voltage sensor i.e. an R1135H mutation which was found in two patients in separate families and a novel R1135C mutation identified in a third patient in another family. Muscle fibres from a patient harbouring the R1135H mutation showed increased depolarization tendency at normal and reduced extracellular potassium compatible with the diagnosis. Additionally, amplitude and rise time of action potentials were reduced compared with controls, even for holding potentials at which all NaV1.4 are fully recovered from inactivation. These findings may be because of an outward omega current activated at positive potentials. Expression of R1135H/C in mammalian cells indicates further gating defects that include significantly enhanced entry into inactivation and prolonged recovery that may additionally contribute to action potential inhibition at the physiological resting potential. After S4 immobilization in the outward position, mutant channels produce an inward omega current that most likely depolarizes the resting potential and produces the hypokalaemia-induced weakness. Gating current recordings reveal that mutations at R3 inhibit S4 deactivation before recovery, and molecular dynamics simulations suggest that this defect is caused by disrupted interactions of domain III S2 countercharges with S4 arginines R2 to R4 during repolarization of the membrane. This work reveals a novel mechanism of disrupted S4 translocation for hypokalaemic periodic paralysis mutations at arginine residues located below the gating pore constriction of the voltage sensor module.
[Show abstract][Hide abstract] ABSTRACT: Purpose:
To implement chlorine 35 ((35)Cl) magnetic resonance (MR) at a 7-T whole-body MR system and evaluate its feasibility for imaging humans.
Materials and methods:
All examinations were performed with ethical review board approval; written informed consent was obtained from all volunteers. Seven examinations each of brain and muscle in healthy volunteers and four examinations of patients were performed. Two patients with histologically confirmed glioblastoma multiforme underwent brain imaging. (35)Cl MR and (35)Cl inversion-recovery (IR) MR were performed. Two patients with genetically confirmed hypokalemic periodic paralysis underwent calf muscle imaging. Seven multiecho sequences (acquisition time, 5 minutes; voxel dimension, 11 mm(3)) were applied to determine transverse relaxation time as affected by magnetic field heterogeneity (T2*) and chlorine concentration. (35)Cl and sodium 23 ((23)Na) MR were conducted with a 7-T whole-body MR system. (35)Cl longitudinal relaxation time (T1) and T2* of healthy human brain and muscle were determined with a three-dimensional density-adapted-projection reconstruction technique to achieve short echo times and high signal-to-noise ratio (SNR) efficiency. A nonlinear least squares routine and mono- (T1) and biexponential (T2*) models were used for curve fitting.
Phantom imaging revealed 15-fold lower SNR and much shorter relaxation times for (35)Cl than (23)Na. In vivo T2* was biexponential and extremely short. Monoexponential fits of T1 revealed 9.2 and 4.0 milliseconds ± 0.7 (standard deviation) for brain and muscle, respectively. In glioblastoma tissue, increased Cl(-) concentrations and increased Cl(-) IR signal intensities were detected. Voxel dimension and acquisition time, respectively, were 6 mm(3) and 9 minutes 45 seconds ((35)Cl MR) and 10 mm(3) and 10 minutes ((35)Cl IR MR). In patients with hypokalemic periodic paralysis versus healthy volunteers, Cl(-) and Na(+) concentrations were increased. Cl(-) concentration of muscle could be determined (voxel size, 11 mm(3); total acquisition time, 35 minutes).
MR at 7 T enables in vivo imaging of (35)Cl in human brain and muscle in clinically feasible acquisition times (10-35 minutes) and voxel volumes (0.2-1.3 cm(3)). Pathophysiological changes of Cl(-) homeostasis due to cancer or muscular ion channel disease can be visualized.
[Show abstract][Hide abstract] ABSTRACT: Malignant hyperthermia (MH) is a rare pharmacogenetic disorder which is characterized by life-threatening metabolic crises during general anesthesia. Classical triggering substances are volatile anesthetics and succinylcholine (SCh). The molecular basis of MH is excessive release of Ca2+ in skeletal muscle principally by a mutated ryanodine receptor type 1 (RyR1). To identify factors explaining the variable phenotypic presentation and complex pathomechanism, we analyzed proven MH events in terms of clinical course, muscle contracture, genetic factors and pharmocological triggers.
In a multi-centre study including seven European MH units, patients with a history of a clinical MH episode confirmed by susceptible (MHS) or equivocal (MHE) in vitro contracture tests (IVCT) were investigated. A test result is considered to be MHE if the muscle specimens develop pathological contractures in response to only one of the two test substances, halothane or caffeine. Crises were evaluated using a clinical grading scale (CGS), results of IVCT and genetic screening. The effects of SCh and volatile anesthetics on Ca2+ release from sarcoplasmic reticulum (SR) were studied in vitro.
A total of 200 patients met the inclusion criteria. Two MH crises (1%) were triggered by SCh (1 MHS, 1 MHE), 18% by volatile anesthetics and 81% by a combination of both. Patients were 70% male and 50% were younger than 12 years old. Overall, CGS was in accord with IVCT results. Crises triggered by enflurane had a significantly higher CGS compared to halothane, isoflurane and sevoflurane. Of the 200 patients, 103 carried RyR1 variants, of which 14 were novel. CGS varied depending on the location of the mutation within the RyR1 gene. In contrast to volatile anesthetics, SCh did not evoke Ca2+ release from isolated rat SR vesicles.
An MH event could depend on patient-related risk factors such as male gender, young age and causative RyR1 mutations as well as on the use of drugs lowering the threshold of myoplasmic Ca2+ release. SCh might act as an accelerant by promoting unspecific Ca2+ influx via the sarcolemma and indirect RyR1 activation. Most MH crises develop in response to the combined administration of SCh and volatile anesthetics.
Full-text · Article · Jan 2014 · Orphanet Journal of Rare Diseases
[Show abstract][Hide abstract] ABSTRACT: A combination of electrophysiological and molecular genetic studies has resulted in the discovery of certain skeletal muscle disorders caused by pathologically functioning ion channels. The group of thus defined hereditary “muscle channelopathies” comprises congenital myasthenic syndromes, non-dystrophic myotonias, dyskalemic periodic paralyses, central-core myopathy and multi-minicore myopathy, as well as malignant hyperthermia. Many muscle channelopathies are benign disorders, but muscle hypermetabolism resulting in muscle stiffness and hyperthermia as in an event of malignant hyperthermia can be life-threatening. Also, forms of familial periodic paralysis can be severe when they produce serious dyskalemia that disturbs cardiac excitation conduction. The hypokalemia is most pronounced in thyrotoxic periodic paralysis. Some of the periodic paralyses are associated with a progressive permanent weakness. The weakness is explained by strongly depolarized, inexcitable muscle fibers that accumulate sodium and water. Drugs that repolarize the fiber membrane can restore muscle strength and may prevent progression. Expression studies of putative mutations have become standard in supporting the disease-causing nature of mutations. Not all variants detected by a genetic analysis may be causative for a clinical dysfunction; they may be just a functional polymorphism. These problems are addressed and a more critical evaluation of the underlying genetic data is proposed.
[Show abstract][Hide abstract] ABSTRACT: Centronuclear myopathy (CNM) is a rare hereditary myopathy characterized by centrally located muscle fiber nuclei. Mutations in the dynamin 2 (DNM2) gene are estimated to account for about 50 % of CNM cases. Electromyographic recordings in CNM may show myopathic motor unit potentials without spontaneous activity at rest. Myotonic discharges, a distinctive electrical activity caused by membrane hyperexcitability, are characteristic of certain neuromuscular disorders. Such activity has been reported in only one CNM case without a known genetic cause. We sequenced the DNM2 gene and the genes associated with myotonia (CLCN1, SCN4A, DMPK and ZNF9) in a sporadic adult patient with CNM and myotonic discharges. Sequencing the entire coding region and exon-intron boundaries revealed a heterozygous c.1106g-a substitution in exon 8, resulting in a R369Q change in the DNM2. Sequencing the CLCN1, SCN4A, DMPK and ZNF9 genes ruled out mutations in these genes. This is the first report of DNM2-related CNM presenting with myotonia. The diagnosis of CNM should be considered in patients with myotonic discharges of an unknown cause.
Full-text · Article · Dec 2013 · Journal of Neural Transmission