RESUMO
The influence of muscle cells on the development of voltage-dependent Ca2+ currents was investigated in Xenopus spinal neurons grown in neuron muscle co-cultures or in muscle-free cultures. Whole-cell currents were separated into low- and high-voltage-activated currents. Developmental changes were assessed by comparing the results obtained at two different periods after plating: 5-10 h (young neurons) and 20-30 h (mature neurons). Our results show a drop in the incidence of low-voltage-activated Ca2+ current with time in both environments: the fraction of young versus mature neurons expressing this current was 67% and 36% in neuron-muscle co-cultures, and 69% and 23% in muscle-free cultures. In both neuron muscle and muscle-free cultures, the density of low-voltage-activated Ca2+ current (when expressed) did not change during the development. In contrast, the density of high-voltage-activated Ca2+ currents increased more than two-fold during the first 30 h in neuron muscle co-cultures, but remained unchanged in muscle-free cultures. This difference was not related to neuronal growth since the increase in neuronal membrane capacitance with time was similar in the two environments. In addition, direct cell-cell interaction through the establishment of functional neuron-muscle synaptic contacts did not further modify the overall expression of high-voltage-activated Ca2+ currents. In conclusion, these results suggest the presence of diffusible factors in neuron muscle co-cultures which up-regulate the expression of high-voltage-activated Ca2+ currents during neuronal development, but do not have any effect on low-voltage-activated Ca2+ currents.
Assuntos
Canais de Cálcio/biossíntese , Regulação da Expressão Gênica no Desenvolvimento , Músculo Esquelético/fisiologia , Neurônios/fisiologia , Medula Espinal/embriologia , Animais , Canais de Cálcio/fisiologia , Células Cultivadas , Técnicas de Cocultura , Embrião não Mamífero , Potenciais da Membrana , Músculo Esquelético/citologia , Neurônios/citologia , Técnicas de Patch-Clamp , Medula Espinal/citologia , Xenopus laevisRESUMO
Functional properties of terminal sprouts elicited by an in vivo injection of Clostridium botulinum type A toxin were studied in endplates of the Levator auris longus muscle of the mouse poisoned from a few days to 28 days beforehand. For this purpose, morphological observations of the extent of terminal sprouts and localization of acetylcholine receptors was performed in whole mount preparations. Sprouts appeared as thin unmyelinated filaments that run usually parallel to the longitudinal axis of the muscle fibres; labelling acetylcholine receptors revealed their line-shaped accumulation co-localized with the sprouts. In addition, presynaptic membrane currents elicited by nerve stimulation were recorded by external electrodes applied under visual control onto the membrane of pre-existing motor endings and newly formed sprouts. These recordings showed the presence of widespread triphasic waveforms which indicated active impulse propagation of the action potential over most of the length of the poisoned endings. Ca2+ influx and Ca2(+)-dependent K+ currents in the sprout membrane were found to be similar to those described in unpoisoned endings. The presence of normal Ca2+ influx, upon active depolarization, in the terminal sprout membranes together with the localization of acetylcholine receptors in front of these membranes, indicates that the terminal sprouts may play a role in the recovery of neuromuscular transmission after Clostridium botulinum poisoning.
Assuntos
Toxinas Botulínicas/toxicidade , Regeneração Nervosa/fisiologia , Junção Neuromuscular/efeitos dos fármacos , 4-Aminopiridina/análogos & derivados , 4-Aminopiridina/farmacologia , Potenciais de Ação/efeitos dos fármacos , Amifampridina , Animais , Canais de Cálcio/efeitos dos fármacos , Canais de Cálcio/metabolismo , Eletrofisiologia , Feminino , Canais Iônicos/efeitos dos fármacos , Camundongos , Terminações Nervosas/efeitos dos fármacos , Terminações Nervosas/metabolismo , Junção Neuromuscular/fisiologia , Junção Neuromuscular/ultraestrutura , Canais de Potássio/efeitos dos fármacos , Receptores Colinérgicos/efeitos dos fármacos , Tetraetilamônio , Compostos de Tetraetilamônio/farmacologiaRESUMO
During synaptogenesis, spontaneous acetylcholine release is rapidly induced by the contact of the growing neurite with the myocyte. Using Xenopus nerve-muscle co-cultures, we investigated the role of Ca2+ in the presynaptic mechanisms mediating this induction process. We found that Ca2+ influx is not necessary to induce spontaneous release and that an increase in cytosolic Ca2+ is not sufficient to trigger this release in the absence of the target cell.
Assuntos
Junção Neuromuscular/metabolismo , Neurotransmissores/metabolismo , Acetilcolina/metabolismo , Compostos de Anilina , Animais , Cádmio/farmacologia , Carbonil Cianeto m-Clorofenil Hidrazona/farmacologia , Células Cultivadas , Citosol/metabolismo , Corantes Fluorescentes , Músculos/citologia , Músculos/efeitos dos fármacos , Músculos/metabolismo , Neuritos/efeitos dos fármacos , Neuritos/metabolismo , Neuritos/fisiologia , Junção Neuromuscular/efeitos dos fármacos , Medula Espinal/citologia , Medula Espinal/efeitos dos fármacos , Sinapses/metabolismo , Sinapses/fisiologia , Xantenos , XenopusRESUMO
This review assembles available information concerning drugs and toxins which block the different types of presynaptic K+ currents and discusses the relative importance of these currents in controlling phasic quantal transmitter release. Drugs and toxins which block the fast voltage-dependent potassium current (IKf), enhance phasic acetylcholine release evoked by nerve impulses. This effect is due to increased Ca2+ influx during prolonged presynaptic membrane depolarization. Selective blockade of the Ca(2+)-dependent K+ current (IK(Ca)) does not induce any change in phasic transmitter release indicating that, under physiological conditions, IK(Ca) has no significant role in presynaptic membrane repolarization. The contribution of the slow voltage-dependent K+ current (IKs) to the regulation of phasic acetylcholine release remains to be clarified. In conclusion, IKf, IK(Ca) and IKs can modulate the entry of Ca2+ into motor nerve terminals. However, under physiological conditions only IKf plays a key role in controlling the transient Ca2+ influx which is responsible for the phasic transmitter release.
Assuntos
Cálcio/metabolismo , Terminações Nervosas/fisiologia , Canais de Potássio/efeitos dos fármacos , Potenciais de Ação/efeitos dos fármacos , Animais , Potenciais da Membrana/efeitos dos fármacosRESUMO
Botulinum toxin type A (Botx) is a potent neurotoxin which inhibits specifically cholinergic synaptic transmission by an unknown mechanism. In order to gain further insight into the mode of action of this toxin, the effect of conditioning nerve stimuli on neuromuscular transmission was studied at endplates of Botx-poisoned and unpoisoned control cutaneous pectoris muscles in the frog. Effects of single conditioning stimuli (facilitation) and multiple high-frequency stimuli (augmentation and potentiation) on epp amplitude and mepp frequency were studied. The main results were that initial facilitation was significantly increased and its decay time constant significantly decreased in Botx-poisoned muscles, while augmentation was unchanged and potentiation was abolished. These changes could be detected before the muscle became completely paralysed, suggesting that they reflect a primary disturbance in the Ca2+-dependent release process.
Assuntos
Toxinas Botulínicas/intoxicação , Cálcio/metabolismo , Junção Neuromuscular/efeitos dos fármacos , Neurotransmissores/metabolismo , Animais , Técnicas In Vitro , Potenciais da Membrana/efeitos dos fármacos , Placa Motora/efeitos dos fármacos , Junção Neuromuscular/metabolismo , Rana temporariaRESUMO
A study of the K conductance of the presynaptic membrane has been performed in the triangularis sterni muscle of the mouse. External currents generated in the presynaptic terminals upon invasion by action potentials have been recorded using microelectrodes inserted into the perineurium of preterminal nerve bundles. The voltage-dependent K current could be pharmacologically dissected into fast (IKf) and slow (IKs) components. While both are sensitive to 3,4-diaminopyridine (3,4-DAP), only IKf is sensitive to tetraethylammonium (TEA). Uranyl (100-200 microM) and guanidine (5-10 mM) produced a near complete block of IKf and IKs, which can explain their facilitatory effect upon transmitter release. The third K current of presynaptic terminals is Ca2+-dependent, but was activated also by Sr2+. This current could be suppressed by nanomolar doses of charybdotoxin; it is also sensitive to TEA but not to 3,4-DAP, uranyl or guanidine.
Assuntos
4-Aminopiridina/análogos & derivados , Junção Neuromuscular/fisiologia , Canais de Potássio/fisiologia , Amifampridina , Aminopiridinas/farmacologia , Animais , Guanidinas/farmacologia , Camundongos , Junção Neuromuscular/efeitos dos fármacos , Compostos Organometálicos/farmacologia , Canais de Potássio/efeitos dos fármacos , Compostos de Tetraetilamônio/farmacologiaRESUMO
1. The effects of botulinum toxin (BoTx) types A and D on spontaneous and evoked phasic transmitter release were studied in the isolated extensor digitorum longus muscle of the rat or the levator auris longus muscle of mice. 2. The toxins were injected subcutaneously into the hindleg of adult rats or the dorsal aspect of the neck of mice. At various times after the injection the muscles were removed from the anaesthetized animal and neuromuscular transmission examined in vitro by conventional intracellular techniques. 3. Both toxins reduced spontaneous transmitter release recorded as the frequency of miniature end-plate potentials but BoTx type D was less effective in that respect than the type A toxin. 4. With both toxins the block of evoked phasic transmitter release, recorded as end-plate potentials, was almost complete. As previously reviewed by Simpson (1986) the block produced by BoTx type A was partially reversed by procedures which elevate the intraterminal level of calcium ions. However, in BoTx type D-paralysed muscles such procedures failed to restore phasic transmitter release but caused a period of high-frequency asynchronous transmitter release following each nerve impulse. 5. To investigate if the lack of synchronization of evoked transmitter release observed in BoTx type D-paralysed muscles was due to alterations in presynaptic currents we examined, by perineural recordings, the Na+, fast K+, slow K+, K+-Ca2+-dependent and the Ca2+ currents in BoTx type D-paralysed muscles. These presynaptic currents were not altered as compared to unpoisoned controls. 6. We suggest that there exists a presynaptic process, which in addition to Ca2+ influx participates in transmitter synchronization and which is a main target for BoTx type D action.
Assuntos
Toxinas Botulínicas/farmacologia , Placa Motora/efeitos dos fármacos , Junção Neuromuscular/efeitos dos fármacos , 4-Aminopiridina/análogos & derivados , 4-Aminopiridina/farmacologia , Potenciais de Ação/efeitos dos fármacos , Amifampridina , Animais , Feminino , Masculino , Camundongos , Junção Neuromuscular/fisiologia , Neurotransmissores/metabolismo , Ratos , Ratos Endogâmicos , Transmissão Sináptica/efeitos dos fármacos , Tetraetilamônio , Compostos de Tetraetilamônio/farmacologiaRESUMO
1. To study the nature and origin of slow-rising, Ca2+-insensitive miniature end-plate potentials (m.e.p.p.s) in mammalian muscle we used intracellular recording techniques and drugs which block acetylcholine (ACh) synthesis or the uptake of ACh into synaptic vesicles. Slow m.e.p.p.s were induced in vivo by paralysing the extensor digitorum longus muscle of the rat with botulinum toxin type A or in vitro by the application of 4-aminoquinoline to the mouse diaphragm nerve-muscle preparation. 2. Hemicholinium-3, which blocks ACh synthesis, reduced the amplitude of all synaptic potentials including slow m.e.p.p.s, but only if the nerve was stimulated. 3. 2(4-phenylpiperidino)cyclohexanol (AH-5183), which blocks the active uptake of ACh into synaptic vesicles, reduced both the frequency and the amplitude of slow m.e.p.p.s and did so without requiring nerve stimulation. 4. No correlation was observed between the molecular leakage of ACh from the motor nerve and the frequency and amplitude of slow m.e.p.p.s. 5. We conclude that slow m.e.p.p.s are caused by the release of ACh from the nerve terminal, possibly from a small pool of synaptic vesicle-like structures.
Assuntos
Cálcio/fisiologia , Placa Motora/fisiologia , Junção Neuromuscular/fisiologia , Piperidinas , Acetilcolina/metabolismo , Potenciais de Ação/efeitos dos fármacos , Aminoquinolinas/farmacologia , Animais , Toxinas Botulínicas/farmacologia , Hemicolínio 3/farmacologia , Técnicas In Vitro , Camundongos , Fármacos Neuromusculares Despolarizantes/farmacologia , Fenciclidina/análogos & derivados , Fenciclidina/farmacologia , Ratos , Ratos Endogâmicos , Temperatura , Fatores de TempoRESUMO
A missense variant (R83H) of the gene (KCNE3) encoding a potassium channel-associated peptide, MinK-related peptide 2 (MiRP2), has been reported in periodic paralysis patients. In the current study, no difference in the frequency of the MiRP2-R83H variant between periodic paralysis patients and healthy individuals was found. Furthermore, there was no segregation of this gene variant with the disease. These observations weaken the proposal that MiRP2-R83H causes periodic paralysis.
Assuntos
Substituição de Aminoácidos , Mutação de Sentido Incorreto , Paralisias Periódicas Familiares/genética , Mutação Puntual , Canais de Potássio de Abertura Dependente da Tensão da Membrana , Canais de Potássio/genética , Adulto , Criança , Segregação de Cromossomos , Feminino , Humanos , Paralisia Periódica Hipopotassêmica/genética , Masculino , Canal de Sódio Disparado por Voltagem NAV1.4 , Paralisia Periódica Hiperpotassêmica/genética , Linhagem , Polimorfismo Conformacional de Fita Simples , Canais de Potássio/química , Canais de Sódio/genética , Tireotoxicose/sangue , Tireotoxicose/complicaçõesRESUMO
A variety of pharmacologically active compounds was surveyed for effects on the Ca2+-insensitive miniature end-plate potentials (slow mepps) induced by botulinum toxin type A (Botx) poisoning in rat muscle. The purpose was to gain insight into the release process responsible for this type of acetylcholine secretion. It was found that caffeine and dibutyryl cyclic adenosine 3',5'-monophosphate increased significantly the frequency of slow mepps in Botx-poisoned muscles, but had no effect on slow mepps in control muscles. Vinblastine and cytochalasin B significantly increased the slow mepp frequency in Botx-poisoned as well as in normal control muscles. Inhibitors of oxidative metabolism reduced the frequency of slow mepps by 90%, indicating a high energy requirement for this type of release. No agent was found to augment the slow mepp frequency above 1-2 Hz, suggesting that an upper limit exists for this type of packaging and release of acetylcholine.
Assuntos
Acetilcolina/metabolismo , Cálcio/fisiologia , Junção Neuromuscular/efeitos dos fármacos , Citoesqueleto de Actina/efeitos dos fármacos , Animais , Calmodulina/metabolismo , Masculino , Potenciais da Membrana/efeitos dos fármacos , Microtúbulos/efeitos dos fármacos , Placa Motora/efeitos dos fármacos , Nucleotídeos Cíclicos/metabolismo , Oxirredução/efeitos dos fármacos , Ratos , Ratos EndogâmicosRESUMO
1. Electrophysiological detection of acetylcholine (ACh) release by synaptosomes from the electric organ of Torpedo was searched for by laying the isolated nerve terminals on a culture of Xenopus embryonic muscle cells (myocytes), and by recording the ACh-induced inward currents in the myocytes. 2. Whole-cell recording in one of the myocytes revealed rapid inward currents that where generated soon after synaptosome application. These pulsatile events strongly resembled those occurring normally during the early phase of synaptogenesis after nerve-muscle contact in Xenopus cell cultures. They were called spontaneous synaptic currents (SSCs). 3. The SSCs produced by the synaptosomes had a rapid time course, with mean time-to-peak and half-decay times of 2.6 +/- 0.4 ms and 6.0 +/- 1.1 ms, respectively. Most events had a falling phase that could be fitted with a single exponential. The mean time constant of decay was 6.2 +/- 1.1 ms. More than half of the SSCs (approximately 60%) constituted a rather homogenous population in which the time-to-peak versus amplitude showed a positive relationship, the smallest events displaying a shorter time course. The rest of the SSCs had a more variable and slower time course. Such events are also observed in young and mature junctions in situ. 4. The amplitudes of SSCs had a wide distribution which was skewed towards the smallest values. The mean amplitude was 65.2 +/- 16.1 pA. 5. During the minutes following an application of synaptosomes, the frequency of the SSCs tended to decrease, but their mean amplitude remained constant. Such behaviour could be reproduced during several successive additions of synaptosomes while recording in the same myocyte. 6. Just after synaptosome application, the SSCs were superposed to a noisy inward current that lasted for 20-60 s. Noise analysis of this current gave the values of 0.7 +/- 0.1 pA for the mean amplitude of the elementary event, and 4.7 +/- 0.2 ms for its mean duration, values that compare well with those reported for the activation of frog embryonic nicotinic receptor. This suggests that the noisy current was due to ACh molecules set free by synaptosomes which were either damaged or which released ACh at some distance. This view was strengthened by biochemical analysis of ACh release by synaptosomes in vitro. 7. Tubocurarine reversibly abolished the appearance of both the noise and the synaptosome-generated SSCs, showing that these currents were due to the action of ACh.(ABSTRACT TRUNCATED AT 400 WORDS)
Assuntos
Acetilcolina/metabolismo , Órgão Elétrico/metabolismo , Sinaptossomos/metabolismo , Torpedo/metabolismo , Animais , Bioensaio , Células Cultivadas , Eletrofisiologia , Cinética , XenopusRESUMO
This study tested the possibility of developing an experimental model of neuropathy in female Wistar rats inoculated with Mycobacterium leprae in the foot pad and assessed by repeated electrophysiological methods. M. leprae multiplied in the rats but considerably less than in simultaneously inoculated mice. No acid-fast bacilli were found in nerves. Motor and sensory conduction velocities remained normal at the thigh level of the sciatic nerve. At the leg, they decreased significantly bilaterally for motor conduction and in the inoculated side for sensory conduction at 21 months after inoculation. These results suggest the possibility of developing an experimental model of leprosy neuropathy which might be useful for therapeutic research. Further histopathological studies are needed to assess this paucibacillary model.
Assuntos
Mycobacterium leprae/crescimento & desenvolvimento , Nervo Isquiático/fisiopatologia , Animais , Eletrofisiologia , Feminino , Pé , Hanseníase/fisiopatologia , Condução Nervosa , Ratos , Ratos EndogâmicosRESUMO
1. The Ile693Thr mutation of the skeletal muscle Na+ channel alpha-subunit is associated with an unusual phenotype of paramyotonia congenita characterized by cold-induced muscle weakness but no stiffness. This mutation occurs in the S4-S5 linker of domain II, a region that has not been previously implicated in paramyotonia congenita. 2. The Ile693Thr mutation was introduced into the human skeletal muscle Na+ gene for functional expression in human embryonic kidney (HEK) cells. The currents expressed were recorded with the whole-cell voltage-clamp technique. 3. In comparison with wild-type currents, Ile693Thr mutant currents showed a clear shift of about -9 mV in the voltage dependence of activation. 4. In contrast to other mutations of the Na+ channel known to cause paramyotonia congenita, the Ile693Thr mutation did not induce any significant change in the kinetics, nor in the voltage dependence, of fast inactivation. 5. In conclusion, this study provides further evidence of the involvement of the S4-S5 linker in the voltage dependence of Na+ channel activation. The negative shift in the voltage dependence found in this mutation must be associated to other defects, plausibly an impairment of the slow inactivation, to account for the long periods of muscle weakness experienced by the patients.
Assuntos
Isoleucina , Músculo Esquelético/metabolismo , Miotonia Congênita/genética , Mutação Puntual , Canais de Sódio/biossíntese , Canais de Sódio/genética , Treonina , Linhagem Celular , Primers do DNA , Humanos , Rim , Potenciais da Membrana/efeitos dos fármacos , Mutagênese Sítio-Dirigida , Técnicas de Patch-Clamp , Potássio/farmacologia , Proteínas Recombinantes/biossíntese , Canais de Sódio/fisiologia , Temperatura , TransfecçãoRESUMO
In addition to motor and sensory nerve conduction, monosynaptic triceps surae reflex, amplitude of median nerve voltage and response of extensor digitorum brevis and soleus muscles were investigated in 29 diabetic patients. These parameters were altered earlier than motor and sensory nerve conduction, which considerably increased the incidence of subclinical peripheral neuropathy (90% of the patients). It was also found that sensory fibers were affected earlier than motor fibers. The electrically detected neuropathy correlated with duration of diabetes and control of glycaemia.
Assuntos
Neuropatias Diabéticas/diagnóstico , Axônios/fisiologia , Neuropatias Diabéticas/fisiopatologia , Eletrofisiologia , Feminino , Humanos , Masculino , Nervo Mediano/fisiopatologia , Pessoa de Meia-Idade , Condução Nervosa , Reflexo Monosináptico , Degeneração Retrógrada , Fatores de TempoRESUMO
1. In the present paper we review some presynaptic aspects of the mode of action of botulinal toxins (BoTxs) at vertebrate neuromuscular junctions with emphasis on studies carried out in our laboratories using electrophysiological and morphological techniques. 2. Spontaneous quantal transmitter release recorded as miniature end-plate potentials is drastically affected by BoTxs. The low probability of release at poisoned terminals can be enhanced by carbonyl cyanide m-chlorophenylhydrazone (CCCP), Cd2+ and La3+. However, CCCP and La3+ which drastically deplete clear synaptic vesicles from unpoisoned terminals failed to markedly affect the density of synaptic vesicles at poisoned terminals. It is concluded that poisoned terminals have a reduced sensitivity to the release-promoting action of Ca2+, Cd2+ and La3+. 3. When comparing the effect of the various BoTxs on nerve-impulse evoked transmitter release it appears that increasing phasic Ca2+ entry into the terminals enhances evoked synchronized quantal release only from terminals poisoned with serotypes A and E. In contrast, enhanced Ca2+ entry into terminals poisoned with serotypes B, D and F induced a period of high frequency asynchronous release suggesting that these BoTxs may affect a presynaptic step beyond the influx of Ca2+, that may be involved in the synchronization of transmitter quanta. These data suggest that the actions of BoTxs involve several steps of the acetylcholine release process. 4. The analysis of presynaptic currents which depend on both Ca2+ entry and intraterminal background Ca2+ levels strongly suggests that neither Ca2+ entry nor intraterminal Ca2+ levels are altered by BoTxs. Furthermore, poisoned terminals are no more efficient than unpoisoned ones in dealing with Ca2+ overloads. 5. Finally, the morphological examination of junctions paralysed by BoTx-A indicates that the toxin triggers a particularly important overgrowth of the nerve terminals and suggests that the in vivo functional recovery may occur from an extension of the original nerve terminal arborization and the concomitant remodelling of postsynaptic structures.
Assuntos
Toxinas Botulínicas/toxicidade , Junção Neuromuscular/efeitos dos fármacos , Sinapses/efeitos dos fármacos , Animais , HumanosRESUMO
Hypokalaemic periodic paralysis (hypoPP) is an autosomal dominant muscle disorder characterized by episodic attacks of muscle weakness associated with a decrease in blood potassium levels. Mutations in the gene encoding the skeletal muscle voltage-gated calcium channel alpha-1 subunit (CACNL1A3) account for the majority of cases. Recently, mutations in the gene coding for the skeletal muscle voltage-gated sodium channel alpha subunit (SCN4A) have been reported in a small number of hypoPP families. In order to determine the relative frequency of the CANCL1A3 and SCN4A mutations in a large population of hypoPP patients, and to specify the clinical and pathological features associated with each of them, we searched for mutations in 58 independent hypoPP index cases. We detected the causative mutation in 45 cases: 40 were linked to the CACNL1A3 gene and five to the SCN4A gene. One mutation has not been described before. Some remarkable clinical features were observed in a large hypoPP family carrying an SCN4A mutation: a complete penetrance in men and women, an early age at onset, postcritic myalgias and an increased number and severity of attacks induced by acetazolamide. A muscle biopsy, performed in two members of this family, revealed a peculiar myopathy characterized by tubular aggregates. In contrast, vacuoles were predominant in muscles from hypoPP patients carrying CACNL1A3 mutations. Our findings point to the usefulness of a molecular characterization of hypoPP patients in clinical practice. They also provide new clues for understanding the mechanisms behind functional and structural alterations of the skeletal muscle in hypoPP.
Assuntos
Paralisia Periódica Hipopotassêmica/genética , Músculo Esquelético/fisiopatologia , Mutação Puntual/genética , Canais de Sódio/genética , Adolescente , Adulto , Idade de Início , Canais de Cálcio/genética , Canais de Cálcio/metabolismo , Canais de Cálcio Tipo L , Criança , Códon/genética , Análise Mutacional de DNA , Feminino , Humanos , Paralisia Periódica Hipopotassêmica/patologia , Paralisia Periódica Hipopotassêmica/fisiopatologia , Masculino , Músculo Esquelético/metabolismo , Músculo Esquelético/patologia , Canal de Sódio Disparado por Voltagem NAV1.4 , Linhagem , Fenótipo , Canais de Sódio/metabolismoRESUMO
The autosomal dominant mutation causing myotonic dystrophy (DM1) is a CTG repeat expansion in the 3'-UTR of the DM protein kinase (DMPK) gene. This multisystemic disorder includes myotonia, progressive weakness and wasting of skeletal muscle and extramuscular symptoms such as cataracts, testicular atrophy, endocrine and cognitive dysfunction. The mechanisms underlying its pathogenesis are complex. Recent reports have revealed that DMPK gene haploinsufficiency may account for cardiac conduction defects whereas cataracts may be due to haploinsufficiency of the neighboring gene, the DM-associated homeobox protein (DMAHP or SIX5) gene. Furthermore, mice expressing the CUG expansion in an unrelated mRNA develop myotonia and myopathy, consistent with an RNA gain of function. We demonstrated that transgenic mice carrying the CTG expansion in its human DM1 context (>45 kb) and producing abnormal DMPK mRNA with at least 300 CUG repeats, displayed clinical, histological, molecular and electrophysiological abnormalities in skeletal muscle consistent with those observed in DM1 patients. Like DM1 patients, these transgenic mice show abnormal tau expression in the brain. These results provide further evidence for the RNA trans-dominant effect of the CUG expansion, not only in muscle, but also in brain.