RESUMO
BACKGROUND: Dysferlinopathy is a phenotypically heterogeneous group of hereditary diseases caused by mutations in the DYSF gene. Early contractures are considered rare, and rigid spine syndrome in dysferlinopathy has been previously reported only once. CASE PRESENTATION: We describe a 23-year-old patient with Miyoshi myopathy with a rigid spine and multiple contractures, a rare phenotypic variant. The disease first manifested when the patient was 13 years old, with fatigue of the gastrocnemius muscles and the development of pronounced contractures of the Achilles tendons, flexors of the fingers, and extensors of the toes, followed by the involvement of large joints and the spine. Magnetic resonance imaging revealed signs of connective tissue and fatty replacement of the posterior muscles of the thighs and lower legs. Edema was noted in the anterior and medial muscle groups of the thighs, lower legs, and the multifidus muscle of the back. Whole genome sequencing revealed previously described mutations in the DYSF gene in exon 39 (c.4282 C > T) and intron 51 (c.5785-824 C > T). An immunohistochemical analysis and Western blot showed the complete absence of dysferlin protein expression in the muscle fibers. CONCLUSIONS: This case expands the range of clinical and phenotypic correlations of dysferlinopathy and complements the diagnostic search for spine rigidity.
Assuntos
Contratura , Miopatias Distais , Atrofia Muscular , Distrofia Muscular do Cíngulo dos Membros , Humanos , Adolescente , Adulto Jovem , Adulto , Proteínas de Membrana/genética , Proteínas Musculares/genética , Distrofia Muscular do Cíngulo dos Membros/complicações , Distrofia Muscular do Cíngulo dos Membros/diagnóstico por imagem , Distrofia Muscular do Cíngulo dos Membros/genética , Mutação , Contratura/etiologia , Contratura/genéticaRESUMO
Clinical observations and studies on different animal models of acquired epilepsy consistently demonstrate that blood-brain barrier (BBB) leakage can be an important risk factor for developing recurrent seizures. However, the involved signaling pathways remain largely unclear. Given the important role of thrombin and its major receptor in the brain, protease-activated receptor 1 (PAR1), in the pathophysiology of neurological injury, we hypothesized that PAR1 may contribute to status epilepticus (SE)-induced epileptogenesis and that its inhibition shortly after SE will have neuroprotective and antiepileptogenic effects. Adult rats subjected to lithium-pilocarpine SE were administrated with SCH79797 (a PAR1 selective antagonist) after SE termination. Thrombin and PAR1 levels and neuronal cell survival were evaluated 48h following SE. The effect of PAR1 inhibition on animal survival, interictal spikes (IIS) and electrographic seizures during the first two weeks after SE and behavioral seizures during the chronic period was evaluated. SE resulted in a high mortality rate and incidence of IIS and seizures in the surviving animals. There was a marked increase in thrombin, decrease in PAR1 immunoreactivity and hippocampal cell loss in the SE-treated rats. Inhibition of PAR1 following SE resulted in a decrease in mortality and morbidity, increase in neuronal cell survival in the hippocampus and suppression of IIS, electrographic and behavioral seizures following SE. These data suggest that the PAR1 signaling pathway contributes to epileptogenesis following SE. Because breakdown of the BBB occurs frequently in brain injuries, PAR1 inhibition may have beneficial effects in a variety of acquired injuries leading to epilepsy.
Assuntos
Região CA1 Hipocampal/metabolismo , Região CA1 Hipocampal/fisiopatologia , Receptor PAR-1/metabolismo , Estado Epiléptico/metabolismo , Trombina/metabolismo , Animais , Peso Corporal/efeitos dos fármacos , Região CA1 Hipocampal/efeitos dos fármacos , Região CA1 Hipocampal/patologia , Masculino , Pirróis/farmacologia , Quinazolinas/farmacologia , Ratos , Ratos Wistar , Receptor PAR-1/antagonistas & inibidores , Estado Epiléptico/patologiaRESUMO
In order to assess the radioecological situation created in the area of the location of diversified uranium mining enterprise "Priargunsky Production Mining and Chemical Association" (PIMCU) there was investigated the radioactivity of a number of the compartments of environment, both at the industrial site and beyond it, as well as the volume activity of radon inside the ground and working premises. Radioecological situation in the vicinity of the uranium mines was performed in comparison with the background (fixed reference, control) district, where there is no uranium mining. Performed studies have shown the significant excess content of 226Ra, 232Th, 210Pb, 222Rn in soil, water open water bodies and local foods near uranium mines compared to areas outside the zone of influence of uranium mining that allows to make a conclusion about the significant technogenic pollution of local areas of the plant and adjoining territory.
Assuntos
Indústria Química , Contaminação Radioativa de Alimentos/análise , Mineração , Radônio , Poluentes Radioativos do Solo/análise , Urânio , Poluentes Radioativos da Água/análise , Fenômenos Ecológicos e Ambientais , Monitoramento Ambiental/métodos , Humanos , Federação RussaRESUMO
Seizures during development are a relatively common occurrence and are often associated with poor cognitive outcomes. Recent studies show that early life seizures alter the function of various brain structures and have long-term consequences on seizure susceptibility and behavioral regulation. While many neocortical functions could be disrupted by epileptic seizures, we have concentrated on studying the prefrontal cortex (PFC) as disturbance of PFC functions is involved in numerous co-morbid disorders associated with epilepsy. In the present work we report an alteration of short-term plasticity in the PFC in rats that have experienced early life seizures. The most robust alteration occurs in the layer II/III to layer V network of neurons. However short-term plasticity of layer V to layer V network was also affected, indicating that the PFC function is broadly influenced by early life seizures. These data strongly suggest that repetitive seizures early in development cause substantial alteration in PFC function, which may be an important component underlying cognitive deficits in individuals with a history of seizures during development.
Assuntos
Plasticidade Neuronal/fisiologia , Córtex Pré-Frontal/fisiopatologia , Convulsões/fisiopatologia , Transmissão Sináptica/fisiologia , Animais , Animais Recém-Nascidos , Técnicas de Patch-Clamp , Ratos , Ratos Sprague-Dawley , Convulsões/complicaçõesRESUMO
Activities related to the rehabilitation of areas and facilities of the temporary storage of spent nuclear fuel and radioactive waste (SNF and RW) at Andreeva Bay and Gremikha on the Kola Peninsula and in the Primorsky Krai in the Russian Far East is an important component of the regulatory functions of the Federal Medical biological Agency (FMBA of Russia). Technical support to the FMBA of Russia in this activity is provided by A.L Burnazyan Federal Medical Biophysical Center Main research interests include evaluation of radiological threats to determine the priority directions of regulation, a detailed analysis of the radiation situation at areas, territories and in vicinity of temporary waste storage facilities, radiation control and environmental monitoring, the development of digital maps and geoinformation systems, project expertise in the field of rehabilitation of PVC including the management of SNF and RW Implementation of these natural, practical and theoretical works is completed by development a set of regulatory documents ensuring adherence to radiation safety for the stuff population and the environment, and the also documents governing the management of SNF and RW waste in the territories of PVC.
Assuntos
Instalações Militares/normas , Monitoramento de Radiação , Proteção Radiológica/métodos , Resíduos Radioativos , Regulamentação Governamental , Higiene Militar , Doses de Radiação , Monitoramento de Radiação/legislação & jurisprudência , Monitoramento de Radiação/métodos , Proteção Radiológica/legislação & jurisprudência , Resíduos Radioativos/análise , Resíduos Radioativos/legislação & jurisprudência , Resíduos Radioativos/prevenção & controle , Federação RussaRESUMO
BACKGROUND: Dysferlinopathy has a high prevalence in relatively isolated ethnic groups where consanguineous marriages are characteristic and/or the founder effect exists. However, the frequency of endemic mutations in most isolates has not been investigated. METHODS: The prevalence of the pathological DYSF gene variant (NM_003494.4); c.200_201delinsAT, p. Val67Asp (rs121908957) was investigated in an isolated Avar population in the Republic of Dagestan. Genetic screenings were conducted in a remote mountainous region characterized by a high level of consanguinity among its inhabitants. In total, 746 individuals were included in the screenings. RESULTS: This pathological DYSF gene variant causes two primary phenotypes of dysferlinopathy: limb-girdle muscular dystrophy (LGMD) type R2 and Miyoshi muscular dystrophy type 1. Results indicated a high prevalence of the allele at 14% (95% confidence interval [CI]: 12-17; 138 out of 1518 alleles), while the allele in the homozygous state was detected in 29 cases-3.8% (CI: 2.6-5.4). The population load for dysferlinopathy was 832.3 ± 153.9 per 100,000 with an average prevalence of limb-girdle muscular dystrophies ranging from 0.38 ± 0.38 to 5.93 ± 1.44 per 100,000. CONCLUSION: A significant burden of the allele was due to inbreeding, as evidenced by a deficiency of heterozygotes and the Wright fixation index equal to 0.14 (CI 0.06-0.23).
RESUMO
There has been a spectacular rise in the global prevalence of type 2 diabetes mellitus and cardiovascular complications are the major cause of morbidity and mortality in diabetic patients. The objective of the study was to investigate ventricular myocyte shortening, intracellular Ca(2+) signalling and expression of genes encoding cardiac muscle proteins in the aged Zucker diabetic fatty (ZDF) rat. There was a fourfold elevation in non-fasting blood glucose in ZDF rats (478.43 ± 29.22 mg/dl) compared to controls (108.22 ± 2.52 mg/dl). Amplitude of shortening, time to peak (TPK) and time to half (THALF) relaxation of shortening were unaltered in ZDF myocytes compared to age-matched controls. Amplitude and THALF decay of the Ca(2+) transient were unaltered; however, TPK Ca(2+) transient was prolonged in ZDF myocytes (70.0 ± 3.2 ms) compared to controls (58.4 ± 2.3 ms). Amplitude of the L-type Ca(2+) current was reduced across a wide range of test potentials (-30 to +40 mV) in ZDF myocytes compared to controls. Sarcoplasmic reticulum Ca(2+) content was unaltered in ZDF myocytes compared to controls. Expression of genes encoding cardiac muscle proteins, membrane Ca(2+) channels, and cell membrane ion transport and intracellular Ca(2+) transport proteins were variously altered. Myh6, Tnnt2, Cacna2d3, Slc9a1, and Atp2a2 were downregulated while Myl2, Cacna1g, Cacna1h, and Atp2a1 were upregulated in ZDF ventricle compared to controls. The results of this study have demonstrated that preserved ventricular myocyte shortening is associated with altered mechanisms of Ca(2+) transport and a changing pattern of genes encoding a variety of Ca(2+) signalling and cardiac muscle proteins in aged ZDF rat.
Assuntos
Sinalização do Cálcio , Tamanho Celular , Diabetes Mellitus Tipo 2/fisiopatologia , Contração Miocárdica , Miócitos Cardíacos/fisiologia , RNA Mensageiro/metabolismo , Animais , Canais de Cálcio Tipo L/metabolismo , Células Cultivadas , Diabetes Mellitus Tipo 2/metabolismo , Expressão Gênica , Masculino , Potenciais da Membrana , Miócitos Cardíacos/metabolismo , RNA Mensageiro/genética , Ratos , Ratos Zucker , Retículo Sarcoplasmático/metabolismoRESUMO
We studied neural induction and generation of neuroectoderm in the colonies of human parthenogenetic SC cultured in the presence of 5 and 19±2% oxygen. We found that neuroectoderm was more actively generated at high oxygen content. At the same time, the transcription of stem cell pluripotency genes was not completely suppressed during neural induction at low oxygen content, while the expression of endoderm and mesodermal marker genes attested to the absence of specific differentiation. These findings demonstrate more efficient neuroectoderm generation induced in the colonies of pluripotent stem cells under conditions of normoxia.
Assuntos
Hipóxia Celular , Placa Neural/citologia , Neurogênese , Células-Tronco Pluripotentes/metabolismo , Proteínas de Ciclo Celular , Diferenciação Celular , Células Cultivadas , Proteínas do Olho/metabolismo , Fatores de Transcrição Forkhead , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Proteína Homeobox Nanog , Fator 3 de Transcrição de Octâmero/genética , Fator 3 de Transcrição de Octâmero/metabolismo , Fator de Transcrição PAX6 , Fatores de Transcrição Box Pareados/metabolismo , Partenogênese , Células-Tronco Pluripotentes/citologia , Proteínas Repressoras/metabolismo , Fatores de Transcrição SOXB1/genética , Fatores de Transcrição SOXB1/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
The association between type 2 diabetes and obesity is very strong, and cardiovascular complications are the major cause of morbidity and mortality in diabetic patients. The aim of this study was to investigate early changes in the pattern of genes encoding cardiac muscle regulatory proteins and associated changes in ventricular myocyte contraction and Ca(2+) transport in young (9- to 13-week-old) type 2 Zucker diabetic fatty (ZDF) rats. The amplitude of myocyte shortening was unaltered; however, time-to-peak shortening and time to half-relaxation of shortening were prolonged in ZDF myocytes (163 ± 5 and 127 ± 7 ms, respectively) compared with age-matched control rats (136 ± 5 and 103 ± 4 ms, respectively). The amplitude of the Ca(2+) transient was unaltered; however, time-to-peak Ca(2+) transient was prolonged in ZDF myocytes (66.9 ± 2.6 ms) compared with control myocytes (57.6 ± 2.3 ms). The L-type Ca(2+) current was reduced, and inactivation was prolonged over a range of test potentials in ZDF myocytes. At 0 mV, the density of L-type Ca(2+) current was 1.19 ± 0.28 pA pF(-1) in ZDF myocytes compared with 2.42 ± 0.40 pA pF(-1) in control myocytes. Sarcoplasmic reticulum Ca(2+) content, release and uptake and myofilament sensitivity to Ca(2+) were unaltered in ZDF myocytes compared with control myocytes. Expression of genes encoding various L-type Ca(2+) channel proteins (Cacna1c, Cacna1g, Cacna1h and Cacna2d1) and cardiac muscle proteins (Myh7) were upregulated, and genes encoding intracellular Ca(2+) transport regulatory proteins (Atp2a2 and Calm1) and some cardiac muscle proteins (Myh6, Myl2, Actc1, Tnni3, Tnn2, and Tnnc1) were downregulated in ZDF heart compared with control heart. A change in the expression of genes encoding myosin heavy chain and L-type Ca(2+) channel proteins might partly underlie alterations in the time course of contraction and Ca(2+) transients in ventricular myocytes from ZDF rats.
Assuntos
Sinalização do Cálcio , Diabetes Mellitus Tipo 2/genética , Diabetes Mellitus Tipo 2/metabolismo , Ventrículos do Coração/metabolismo , Miócitos Cardíacos/metabolismo , Disfunção Ventricular/genética , Disfunção Ventricular/metabolismo , Citoesqueleto de Actina/metabolismo , Animais , Canais de Cálcio/genética , Canais de Cálcio/metabolismo , Diabetes Mellitus Tipo 2/fisiopatologia , Regulação para Baixo , Regulação da Expressão Gênica , Ventrículos do Coração/fisiopatologia , Proteínas Musculares/genética , Proteínas Musculares/metabolismo , Contração Miocárdica/genética , Contração Miocárdica/fisiologia , Miócitos Cardíacos/fisiologia , Cadeias Pesadas de Miosina/genética , Cadeias Pesadas de Miosina/metabolismo , Obesidade/genética , Obesidade/metabolismo , Obesidade/fisiopatologia , Ratos , Ratos Zucker , Retículo Sarcoplasmático/metabolismo , Disfunção Ventricular/fisiopatologiaRESUMO
We studied the effect of recombinant human activin A on induced neuroectoderm formation in colonies of human parthenogenetic SC in the absence of feeder cells. It was found that pretreatment of human parthenogenetic SC with activin A suppressed subsequent neural induction. Activin A in a concentration of 10 ng/ml significantly decreased transcriptional activity of genes required for neuroectoderm formation. At the same time, activin A in a concentration of 20 ng/ml increased the expression of pluripotency genes and completely inhibited the formation of structures in vitro reproducing the neural tube of the developing embryo. These findings attest to prolonged effect of activin A as an inhibitor of neuroectodermal differentiation.
Assuntos
Ativinas/farmacologia , Placa Neural/efeitos dos fármacos , Animais , Células Cultivadas , Regulação da Expressão Gênica/efeitos dos fármacos , Camundongos , Partenogênese , Proteínas Recombinantes/farmacologia , Transcrição Gênica/efeitos dos fármacosRESUMO
Genomic imprinting is one of the most remarkable and important epigenetic phenomena. A biological ban on parthenogenetic and androgenetic development of mammals is an obvious consequence of genomic imprinting. Genomic imprinting defects may cause malformations, clinical syndromes, and tumor growth in humans and to the large offspring syndrome and an increased mortality after in vitro manipulations with early embryos in mammals. Differential expression of parental alleles during ontogeny implies a mechanism of reversible, selective marking of gene alleles. These relatively stable epigenetic modifications, which do not affect the primary nucleotide sequence of DNA, may be transmitted in somatic cell lines and reproduced in the germ line. The genomic imprinting mechanism may be involved in other epigenetic processes, such as epigenetic inheritance, nonrandom allele segregation, meiotic drive, etc. Artificial modulation of genomic imprinting effects with the use of growth factors and demethylating agents permits partial "normocoping" during the development of parthenogenetic mouse embryos. Targeted changes in the transcriptional activity of imprinted genes provide prerequisites for epigenetic correction of syndromes and diseases caused by genomic imprinting defects.
Assuntos
Epigênese Genética , Impressão Genômica , Mamíferos/genética , Animais , Desenvolvimento Embrionário/genética , Camundongos , PartenogêneseRESUMO
The CA3 and CAI regions are the main stages of the "three-synaptic pathway", which plays a role in the generation of hyper-synchronous events in the hippocampus. Under certain experimental conditions, this brain structure might support pathological epileptiform synchronization that is independent of active chemical synaptic transmission. In present work, we estimated the conditions that would facilitate non- synaptic synchronization of the hippocampus. Non-synaptic epileptiform activity was induced in hippocampal slices by the omission calcium ions from the extracellular milieu. The propensity of hippocampal regions to nonsynaptic interactions was estimated by measuring the delay time neededfor the development of low-Ca²âº discharges in the CA3 and CAI. Next, an increase of neuronal excitability was induced by the pre- incubation ofhippocampal slices in 4-aminopyridine (4-AP) and by the reduction ofextracellular osmolarity. Pre-incubation of hippbcampal slices with 4-AP under normal osmotic conditions resulted in decreased latency for non-synaptic discharges in the CA3, but not in the CAl. However hypo-osmotic conditions caused increased excitability of the CA3 region, which resulted in decreased delay time for nonsynaptic discharges and this level of cellular excitability was not further enhanced by the pre-incubation with 4-AR.
Assuntos
Região CA1 Hipocampal/fisiopatologia , Região CA3 Hipocampal/fisiopatologia , Cálcio/metabolismo , Epilepsia/fisiopatologia , Potenciais da Membrana/fisiologia , Modelos Biológicos , Animais , Região CA1 Hipocampal/efeitos dos fármacos , Região CA1 Hipocampal/metabolismo , Região CA3 Hipocampal/efeitos dos fármacos , Região CA3 Hipocampal/metabolismo , Epilepsia/metabolismo , Espaço Extracelular/metabolismo , Potenciais da Membrana/efeitos dos fármacos , Técnicas de Cultura de Órgãos , Pressão Osmótica , Bloqueadores dos Canais de Potássio/farmacologia , Pontuação de Propensão , Ratos Wistar , Transmissão Sináptica/efeitos dos fármacos , Transmissão Sináptica/fisiologiaRESUMO
Decreasing of surface charge screening near voltage-gated ion channels via reduction of extracellular cation divalent ions provide potent mechanism of altering cellular excitability and seizure threshold. Spontaneous field potentials were recorded from horizontal brain slices of young Wistar rats (postnatal day 10-12). Extracellular registrations wereobtained from CA1 and CA3 area of hippocampus. For induction of nonsynaptic epileptiform activity slices were perfused with artificial cerebrospinal fluid with omitted Ca2+and Mg2+ ions. Effect of different Mg2+ concentration (1, 2, and 3mmol/l) on initial stage of nonsynaptic epileptiform discharges was studied. Our results suggest that the change in Mg2+ concentration dramatically affects the probability of induction of low-Ca2+seizure-like activity (SLA), providing evidence that Mg2+ can alter cerebral excitability by affecting the surface charge and supporting the idea that surface charge could be a pharmacological target for anti-epileptic treatment.
Assuntos
Região CA1 Hipocampal/efeitos dos fármacos , Região CA3 Hipocampal/efeitos dos fármacos , Cálcio/metabolismo , Potenciais Evocados/efeitos dos fármacos , Magnésio/farmacologia , Animais , Animais Recém-Nascidos , Materiais Biomiméticos/química , Materiais Biomiméticos/farmacologia , Região CA1 Hipocampal/metabolismo , Região CA1 Hipocampal/fisiopatologia , Região CA3 Hipocampal/metabolismo , Região CA3 Hipocampal/fisiopatologia , Cálcio/farmacologia , Cátions Bivalentes , Líquido Cefalorraquidiano/química , Meios de Cultura/química , Meios de Cultura/farmacologia , Epilepsia/metabolismo , Epilepsia/fisiopatologia , Microtomia , Modelos Biológicos , Técnicas de Patch-Clamp , Ratos , Ratos Wistar , Eletricidade Estática , Técnicas de Cultura de TecidosRESUMO
Acid-sensing ion channels (ASICs) play an important role in numerous functions in the central and peripheral nervous systems ranging from memory and emotions to pain. The data correspond to a recent notion that each neuron and many glial cells of the mammalian brain express at least one member of the ASIC family. However, the mechanisms underlying the involvement of ASICs in neuronal activity are poorly understood. However, there are two exceptions, namely, the straightforward role of ASICs in proton-based synaptic transmission in certain brain areas and the role of the Ca(2+)-permeable ASIC1a subtype in ischaemic cell death. Using a novel orthosteric ASIC antagonist, we have found that ASICs specifically control the frequency of spontaneous inhibitory synaptic activity in the hippocampus. Inhibition of ASICs leads to a strong increase in the frequency of spontaneous inhibitory postsynaptic currents. This effect is presynaptic because it is fully reproducible in single synaptic boutons attached to isolated hippocampal neurons. In concert with this observation, inhibition of the ASIC current diminishes epileptic discharges in a low Mg(2+) model of epilepsy in hippocampal slices and significantly reduces kainate-induced discharges in the hippocampus in vivo Our results reveal a significant novel role for ASICs.This article is part of the themed issue 'Evolution brings Ca(2+) and ATP together to control life and death'.
Assuntos
Bloqueadores do Canal Iônico Sensível a Ácido/farmacologia , Canais Iônicos Sensíveis a Ácido/genética , Epilepsia/fisiopatologia , Hipocampo/efeitos dos fármacos , Canais Iônicos Sensíveis a Ácido/metabolismo , Animais , Hipocampo/metabolismo , Rede Nervosa/metabolismo , RatosRESUMO
The brain slice preparation is the most frequently used tool for testing of pharmacological agents on the neuronal excitability. However in the absence of blood circulation in vitro, the tissue oxygenation strongly depends on the experimental conditions. It is well established that both hypoxia as well as hyperoxia can modulate the neuronal network activity. Thereby changes in tissue oxygen level during experiment may affect the final result. In the present study we investigated the effect of oxygenation on seizure susceptibility in the hippocampal slice preparation using 4-aminopyridine (4-AP) model of ictogenesis in inmature rats. We found that changing the medium perfusion rate in the range of 1-5 ml/min greatly affects the tissue oxygenation, amplitude and frequency of 4-AP-induced synchronous neuronal activity. The decrease in the flow rate as well as substitution of the oxygen in the extracellular medium with nitrogen causes a strong reduction of 4-AP-induced synchronous neuronal discharges. Our results demonstrate a significant linear correlation between the power of 4-AP-induced neuronal activity and the oxygen level in slice tissue. Also we demonstrated that the presence of medium flow is a necessary condition to support the constant level of the slice oxygenation. These data suggest that the oxygen supply of the brain slice strongly depends on experimental protocol and could modulate in vitro neuronal network excitability which should be taken into consideration when planning epilepsy-related studies.
Assuntos
4-Aminopiridina/farmacologia , Hipocampo/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Oxigênio/farmacologia , Bloqueadores dos Canais de Potássio/farmacologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Animais , Animais Recém-Nascidos , Hipóxia Celular , Meios de Cultura/química , Meios de Cultura/farmacologia , Hipocampo/citologia , Hipocampo/fisiologia , Hiperóxia/induzido quimicamente , Hiperóxia/fisiopatologia , Microtomia , Neurônios/citologia , Neurônios/fisiologia , Perfusão/métodos , Ratos , Ratos Wistar , Técnicas de Cultura de TecidosRESUMO
Serine protease thrombin, a key factor of blood coagulation, participates in many neuronal processes important for normal brain functioning and during pathological conditions involving abnormal neuronal synchronization, neurodegeneration and inflammation. Our previous study on CA3 pyramidal neurons showed that application ofthrombin through the activation of specific protease-activated receptor 1 (PAR1) produces a significant hyperpolarizing shift of the activation of the TTX-sensitive persistent voltage-gated Na+ current (I(Nap)) thereby affecting membrane potential and seizure threshold at the network level. It was shown that PAR1 is also expressed in CA1 area of hippocampus and can be implicated in neuronal damage in this area after status epilepticus. The aim of the present study was to evaluate the effect of thrombin on I(NaP) in CA1 pyramidal neurons from adult and young rats. Using whole cell patch-clamp technique we demonstrate that thrombin application results in the hyperpolarization shift of I(NaP) activation as well as increase in the I(NaP) amplitude in both age groups. We have found that I(NaP) in pyramidal neurons of hippocampal CA 1 region is more vulnerable to the thrombin action than I(NaP) in pyramidal neurons of hippocampal CA3 region. We have also found that the immature hippocampus is more sensitive to thrombin action which emphasizes the contribution of thrombin-dependent pathway to the regulation of neuronal activity in immature brain.
Assuntos
Envelhecimento/fisiologia , Região CA1 Hipocampal/efeitos dos fármacos , Região CA3 Hipocampal/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Sódio/metabolismo , Trombina/farmacologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Animais , Região CA1 Hipocampal/citologia , Região CA1 Hipocampal/fisiologia , Região CA3 Hipocampal/citologia , Região CA3 Hipocampal/fisiologia , Expressão Gênica , Microtomia , Neurônios/citologia , Neurônios/fisiologia , Especificidade de Órgãos , Técnicas de Patch-Clamp , Ratos , Ratos Wistar , Receptor PAR-1/genética , Receptor PAR-1/metabolismo , Tetrodotoxina/farmacologia , Técnicas de Cultura de Tecidos , Canais de Sódio Disparados por Voltagem/metabolismoRESUMO
Using an experimental model of neonatal recurrent seizures we investigated the influence of epileptic seizures in the various forms of synaptic plasticity in neurons of the somatosensory cortex. We found that early seizures do not affect the post-tetanic potentiation of the amplitude of the postsynaptic potentials and the depression of postsynaptic potentials during high-frequency stimulation. However they result in the chronic increase of the long-term potentiation of synaptic transmission. These changes of synaptic plasticity may affect the processing of the sensory information in patients with a history of recurrent seizures during early development.
Assuntos
Epilepsia/fisiopatologia , Potenciação de Longa Duração/efeitos dos fármacos , Córtex Somatossensorial/fisiopatologia , Transmissão Sináptica/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Convulsivantes/farmacologia , Epilepsia/induzido quimicamente , Epilepsia/patologia , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Flurotila/farmacologia , Humanos , Potenciais Pós-Sinápticos Inibidores/efeitos dos fármacos , Masculino , Neurônios/efeitos dos fármacos , Neurônios/patologia , Ratos Sprague-Dawley , Córtex Somatossensorial/efeitos dos fármacos , Córtex Somatossensorial/patologiaRESUMO
This study employing a rodent model of neuropathic pain investigated the influence of partial nerve injury on the ability of NMDA receptor activation to induce membrane currents and rises in cytosolic concentration of free calcium ([Ca2+]i) in the rat substantia gelatinosa (SG) neurons using simultaneous whole-cell patch-clamp recording and fura-2 calcium imaging in spinal slices. The novel findings are that: (I) L5-L6 spinal nerve ligation produces a sustained facilitation of NMDA-mediated membrane currents and [Ca2+]i rises both in the soma and dendrites of SG neurons on the injured side on post-operative days 4-13 after injury. (2) It appears that SG neurons in slices from injured rats recover from Ca2+ load less efficiently than neurons from naive rats. (3) The membrane depolarization-induced Ca2+ transients in SG neurons are not modified following spinal nerve ligation. The temporal profile of the changes in Ca2+ transients correlated well with the development of mechanical and thermal allodynia and hyperalgesia. These results suggest an important role of NMDA-mediated calcium signalling in the pathogenesis of neuropathic pain following spinal nerve injury.
Assuntos
Sinalização do Cálcio/fisiologia , Neurônios Aferentes/metabolismo , Dor/metabolismo , Doenças do Sistema Nervoso Periférico/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Substância Gelatinosa/metabolismo , Animais , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Denervação/efeitos adversos , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Vértebras Lombares , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/fisiologia , N-Metilaspartato/farmacologia , Neurônios Aferentes/efeitos dos fármacos , Dor/patologia , Dor/fisiopatologia , Doenças do Sistema Nervoso Periférico/patologia , Doenças do Sistema Nervoso Periférico/fisiopatologia , Ratos , Ratos Sprague-Dawley , Receptores de N-Metil-D-Aspartato/efeitos dos fármacos , Nervos Espinhais/cirurgia , Substância Gelatinosa/efeitos dos fármacos , Substância Gelatinosa/fisiopatologia , Transmissão Sináptica/efeitos dos fármacos , Transmissão Sináptica/fisiologiaRESUMO
The autosomal recessive gene hairless (hr) is responsible for the complete hairlessness in mice homozygous for this gene. Hair shedding that begins at the age of 10 days is caused by an abnormal cycle of hair follicle development disturbed at the catagen stage. This results in enhanced programmed cell death (apoptosis) and ultimately leads to the complete hair follicle destruction and shedding of all hairs by the age of three weeks. To study the phenotypic expression of the hr gene in a chimeric organism, we have obtained 12 chimeric mice hr/hr <--> +/+ by means of aggregation of early embryos hr/hr and +/+. In chimeric mice, the hair shedding has begun two days later than in the hr/hr mice. By day 23 of postnatal development, hairless areas were present on the coat of chimeric mice or the latter were completely hairless depending on the percentage of the hr/hr mutant component. In four chimeras with high content of the mutant component (68-76%), the hair shedding process was similar to that in the hr/hr mice, though it was accomplished two days later. In three chimeras with 48-51% of the mutant component, alternating hairless and hair-covered bands were observed. These data suggest that the hr gene acts in epidermal cells of a hair follicle, because epidermal cell clones in embryonic skin migrate in the lateral-ventral direction coherently and without mixing. However, some chimeras displayed a pattern which was not so clear-cut: the band borders were illegible and hairs partly covered the hairless areas. In some chimeras, the uniform thinning of the coat was observed. Analysis of the effects of the hr mutant gene in chimeric mice differing in the ratio between mutant (hr/hr) and normal (+/+) components in tissues suggests that the hr gene acts in the epidermal cells of the hair follicle. The interactions between cells have an essential effect on the mode and degree of the hr gene expression, which leads to distortion of the "ectodermal" coat pattern in chimeras.
Assuntos
Quimera , Camundongos Pelados/genética , Mutação , Animais , Camundongos , Camundongos Endogâmicos C57BLRESUMO
The mutant gene wellhaaring (we) confers the waved coat in mice, which is most pronounced in homozygotes at 10 to 21 days of postnatal development. Abnormal hair growth and structure in the we/we mutant mice results from defective cell differentiation in the inner root sheath of a hair follicle. To localize the site of the we gene action, we obtained ten chimeric mice by aggregation of the early C57BL/6-2we/we and BALB/c embryos. The chimera coat was waved, shaggy, or almost normal depending on the percentage of the mutant component. In the we/we +/+ chimeric animals of the first generation (G1) aged 21 days, both mutant and normal hair phenotypes were observed, which was especially discernible in zigzag hair. Note that none of the chimeras exhibited the alternating patterns of transversely oriented stripes or patches of either mutant or normal hair; i.e., they had a mixed parental hair phenotype. We also did not observe the animals with an intermediate phenotype, which suggests a discontinuous hair formation in chimeras according to the "all or nothing" principle. The data obtained indicate that the dermal papilla cells of a hair follicle are the sites for the we gene action. During the embryonic development, dermal cells are strongly mixed, which accounts for the lack of the clear-cut transverse stripes of either mutant or normal hair. The mutant gene we is probably responsible for a disrupted induction signal from the dermal papilla towards ectodermal cells of a hair follicle.