Expression changes of NaV channel subunits correlate with developmental maturation of electrophysiological characteristics of rat cerebellar Purkinje neurons / 南方医科大学学报

OBJECTIVE@#To investigate the variations in the expression of voltage-gated sodium (Nav) channel subunits during development of rat cerebellar Purkinje neurons and their correlation with maturation of electrophysiological characteristics of the neurons.@*METHODS@#We observed the changes in the expression levels of NaV1.1, 1.2, 1.3 and 1.6 during the development of Purkinje neurons using immunohistochemistry in neonatal (5-7 days after birth), juvenile (12-14 days), adolescent (21-24 days), and adult (42-60 days) SD rats. Using whole-cell patch-clamp technique, we recorded the spontaneous electrical activity of the neurons in ex vivo brain slices of rats of different ages to analyze the changes of electrophysiological characteristics of these neurons during development.@*RESULTS@#The expression of NaV subunits in rat cerebellar Purkinje neurons showed significant variations during development. NaV1.1 subunit was highly expressed throughout the developmental stages and increased progressively with age (P < 0.05). NaV1.2 expression was not detected in the neurons in any of the developmental stages (P > 0.05). The expression level of NaV1.3 decreased with development and became undetectable after adolescence (P < 0.05). NaV1.6 expression was not detected during infancy, but increased with further development (P < 0.05). NaV1.1 and NaV1.3 were mainly expressed in the early stages of development. With the maturation of the rats, NaV1.3 expression disappeared and NaV1.6 expression increased in the neurons. NaV1.1 and NaV1.6 were mainly expressed after adolescence. The total NaV protein level increased gradually with development (P < 0.05) and tended to stabilize after adolescence. The spontaneous frequency and excitability of the Purkinje neurons increased gradually with development and reached the mature levels in adolescence. The developmental expression of NaV subunits was positively correlated with discharge frequency (r=0.9942, P < 0.05) and negatively correlated with the excitatory threshold of the neurons (r=0.9891, P < 0.05).@*CONCLUSION@#The changes in the expression levels of NaV subunits are correlated with the maturation of high frequency electrophysiological properties of the neurons, suggesting thatmature NaV subunit expressions is the basis of maturation of electrophysiological characteristics of the neurons.

Effect of electromagnetic field from 2g, 3g and 4g mobile phones on the organization of purkinje cell layer of rat cerebellum

Objective: To study and compare the histomorphological changes induced by second generation [2G], third generation [3G] and fourth generation [4G] mobile phone electromagnetic fields on the organization of Purkinje cell layer of rat cerebellum

Study Design: Laboratory based randomized control trial

Place and Duration of Study: The study was carried out in the Anatomy department, Army Medical College Rawalpindi; in collaboration with animal house, National Institute of Health, Islamabad, from Nov 2014 to Nov 2015

Material and Methods: Forty adult Sprague Dawley rats [20 male, 20 female], weighing 250-350grams,were taken and divided into 4 groups with 10 rats [5 male, 5 female] in each group. Group A served as control and was given normal diet and water ad libitum. Groups B, C and D were exposed to EMF from 2G, 3G and 4G mobile phones respectively, daily for 1 hour for 2 months. The animals were sacrificed on 60th day of the experiment, cerebellums were removed, fixed in 10% formalin, processed and stained with haematoxylin and eosin [H and E] for histological study

Results: It was observed on microscopic examination that EMF from 2G, 3G and 4G mobile phones effected the organization of Purkinje cell layer of cerebellum; being double cell layer in group B and multiple cell layers in groups C and D

Conclusion: It was concluded from current results that radiations from 2G, 3G and 4G mobile phones have deleterious effects on the organization of Purkinje cell layer of cerebellum with 3G and 4G causing more harm as compared to EMF from 2G mobile phones

Evaluation of brachial plexus fascicles involvement on infraclavicular block: unfixed cadaver study / Avaliação do envolvimento dos fascículos do plexo braquial no bloqueio por via infraclavicular: estudo em cadáveres não fixados / Evaluación de la implicación de los fascículos del plexo braquial en el bloqueo por vía infraclavicular: estudio en cadáveres no fijos

BACKGROUND AND OBJECTIVES: This study shows how the diffusion of the anesthetic into the sheath occurs through the axillary infraclavicular space and hence proves the efficacy of the anesthetic block of the brachial plexus, and may thereby allow a consolidation of this pathway, with fewer complications, previously attached to the anesthesia. MATERIALS AND METHODS: 33 armpits of adult cadavers were analyzed and unfixed. We injected a solution of neoprene with latex dye in the infraclavicular space, based on the technique advocated by Gusmão et al., and put the corpses in refrigerators for three weeks. Subsequently, the specimens were thawed and dissected, exposing the axillary sheath along its entire length. RESULTS AND DISCUSSION: Was demonstrated involvement of all fasciculus of the plexus in 51.46%. In partial involvement was 30.30%, 18.24% of cases the acrylic was located outside the auxiliary sheath involving no issue. CONCLUSIONS: The results allow us to establish the infraclavicular as an effective and easy way to access plexus brachial, because the solution involved the fascicles in 81.76% partially or totally, when it was injected inside the axillary sheath. We believe that only the use of this pathway access in practice it may demonstrate the efficiency. .

JUSTIFICATIVA E OBJETIVOS: Procuramos demonstrar como ocorre a difusão do anestésico no interior da bainha axilar, quando se utiliza o bloqueio por via infraclavicular, através da fossa infraclavicular e, consequentemente, provar a eficácia dessa via, podendo, com isso, permitir uma consolidação da utilização desse acesso, com redução das complicações. MATERIAS E MÉTODO: Foram utilizadas 33 axilas de cadáveres adultos não fixados. Injetamos uma solução de neoprene látex com corante na fossa infraclavicular, baseando-se na técnica preconizada por Gusmão e col, e colocamos os cadáveres em geladeiras por três semanas. Posteriormente, as peças foram descongeladas e dissecadas, expondo a bainha axilar em toda sua extensão. RESULTADOS E DISCUSSÃO: Foi demonstrado envolvimento de todos os fascículos do plexo em 51,46%. Em 30,30% houve envolvimento parcial, e em 18,24% dos casos o acrílico foi localizado fora da bainha axilar, não envolvendo nenhum fascículo. CONCLUSÕES: Os dados obtidos permitem estabelecer a via infraclavicular como uma via eficaz e de fácil acesso ao plexo braquial, visto que a solução injetada envolveu os fascículos em 81,76% parcialmente ou totalmente, quando era injetada dentro da bainha axilar. Acreditamos que apenas a utilização desta via de acesso na prática poderá demonstrar a eficiência da mesma. .

JUSTIFICACIÓN Y OBJETIVOS: Este estudio intenta demostrar cómo ocurre la difusión del anestésico en el interior de la vaina axilar, cuando se utiliza el bloqueo por vía infraclavicular a través de la fosa infraclavicular, y al mismo tiempo, probar la eficacia de esa vía, pudiendo así permitir una consolidación de la utilización de ese acceso con reducción de las complicaciones. MATERIALES Y MÉTODO: Fueron utilizadas 33 axilas de cadáveres adultos no fijadas. Inyectamos una solución de neopreno látex con colorante en la fosa infraclavicular, con la técnica preconizada por Gusmão et al., y colocamos los cadáveres en frigoríficos durante 3 semanas. Posteriormente, las piezas fueron descongeladas y disecadas, exponiendo la vaina axilar en toda su extensión. RESULTADOS Y DISCUSIÓN: Quedó demostrada la implicación de todos los fascículos del plexo en un 51,46%. En un 30,30% hubo una participación parcial, y en un 18,24% de los casos el acrílico fue ubicado fuera de la vaina axilar sin la participación de ningún fascículo. CONCLUSIONES: Los datos obtenidos permiten establecer la vía infraclavicular como una vía eficaz y de fácil acceso al plexo braquial, visto que la solución inyectada tuvo la participación de los fascículos en un 81,76% parcial o totalmente, cuando se inyectaba dentro de la vaina axilar. Creemos que solamente con la utilización de esta vía de acceso en la práctica podrá quedar demostrada su eficacia. .

Morphology of the atrioventricular junction in Iguana iguana (Reptilia-Iguanidae)

The atrioventricular junctional area (AVJA), including atrioventricular(AV) node and bundle was investigated in seven hearts of common or green iguana (Iguana iguana) using the light microscopy. Adult animals, both sexes, were captured in the Pantanal, Brazil. All hearts were fixed in buffered formaldehyde 10 (pH 7.2) for 24 hours, embedded in paraplast according to routine methods, and serially cutat 5 ?m thickness. In the Iguana iguana, the AVJA consists of a massof the fibers intermingled with variable amount of connective tissue and blood vessels surrounded by adjacent myocardium and theattachment of the right atrioventricular valve in the fibrous skeleton. By light microscopy, conducting cells of the AV node and bundle canbe distinguished from working cells by their much smaller size, palerstaining reaction and the presence of a sheath of connective tissue.The AV node and bundle and its branches were found to constitutea continuous tract. Histochemically, we found elastic fibers between cells of the conduction, mainly in the AV node. The PAS method reveals absence of glycogen in specialized cells. The fibrous skeleton, mainly the right trigone, showed a well-developed chondroid tissue,made by hyaline like cartilage (binucleated condrocytes included in thebig lacunas and extracellular matrix with fibrillar collagen). In conclusion, the nodal and Purkinje cells in heart iguana presented poorly morphological differentiation comparing mammals and birds, however the skeleton fibrous has a different cartilage kind.

A área da junção atrioventricular (AJAV), incluindo nó e feixe atrioventriculares foi investigada em sete corações de iguana comum ou verde (Iguana iguana), usando microscopia de luz. Animais adultos, de ambos os sexos, foram capturados no Pantanal, Brasil. Todos os corações foram fixados em formaldeído tamponado 10 (pH 7,2) por 24 horas, incluídos em parapast de acordo com métodos de rotina e seccionados com 5?m de espessura. Na iguana, a AJAV consiste de uma massa de fibras musculares mergulhada em uma quantidade variável de tecido conjuntivo e vasos sangüíneos, rodeada por miocárdio adjacente e aderida à valva atrioventricular direita no esqueleto fibroso. Através da microscopia de luz, células de conduçãodo nó e feixe atrioventriculares podem ser distinguidas das células de trabalho por serem menores, de coloração pálida e pela presença de um envoltório de tecido conjuntivo. O nó AV bem como o feixe AVe seus ramos formam um trato contínuo. Histologicamente, encontramos fibras elásticas entre as células de condução, principalmente no nó AV. O método do PAS revelou a presença de glicogênio nas células especializadas. O esqueleto fibroso, principalmente, o trígono direito, apresentou um tecido condróide bem desenvolvido, constituído de cartilagem semelhante à hialina (condrócitos binucleados inseridos em grandes lacunas e matriz extracelular com colágeno fibrilar). O esqueleto fibroso tem fibrascolágenas e cartilagem semelhante à hialina. Em conclusão, as células nodais e de Purkinje no coração de iguana apresentam pouca diferença morfológica quando comparadas às de mamíferos e aves, contudo o esqueleto fibroso tem um tipo diferente de cartilagem.

Immunoelectronic localization of myosin-Va in rat cerebellum

Myosin V is an unconventional type of actin-binding myosin that participates in cytoplasmic organelle transport. Although this unconventional myosin has been extensively studied, its subcellular localization in the mammalian cerebellum has not been determined. In this work, we used an antibody against the tail domain of the myosin-Va heavy chain and a secondary antibody labeled with protein A-gold (15 nm) to study the subcellular distribution of this protein. Myosin-Va was found in the cytoplasm, where it was associated with a filament (probably actin). This protein was also detected in the plasma membrane of axons and dendrites in the molecular layer in rat cerebellum.

Further observations on cerebellar climbing fibers. A study by means of light microscopy, confocal laser scanning microscopy and scanning and transmission electron microscopy

The intracortical pathways of climbing fibers were traced in several vertebrate cerebella using light microscopy, confocal laser scanning microscopy, scanning and transmission electron microscopy. They were identified as fine fibers up to 1(micron thick, with a characteristic crossing-over bifurcation pattern. Climbing fiber collaterals were tridimensionally visualized forming thin climbing fiber glomeruli in the granular layer. Confocal laser scanning microscopy revealed three types of collateral processes at the interface between granular and Purkinje cell layers. Scanning electron microscopy showed climbing fiber retrograde collaterals in the molecular layer. Asymmetric synaptic contacts of climbing fibers with Purkinje dendritic spines and stellate neuron dendrites were characterized by transmission electron microscopy. Correlative microscopy allowed us to obtain the basic three-dimensional morphological features of climbing fibers in several vertebrates and to show with more accuracy a higher degree of lateral collateralization of these fibers within the cerebellar cortex. The correlative microscopy approach provides new views in the cerebellar cortex information processing.

4-Aminopyridine (4-AP) augments Ca(2+)-dependent action potential and changes oscillatory firing patterns in rat cerebellar Purkinje cells

Intracellular recordings in cerebellar slice preparation showed that applications of 4-AP altered the pattern of oscillatory firing activity in Purkinje cells (PCs), especially yielding pronounced changes in action potential shape. 4-AP increased the amplitude and duration of action potential significantly and decreased the spike frequency. After 4-AP application, the duration of bursting was prolonged and the duration of after-burst hyperpolarization was progressively shortened. In all PCs tested, the rhythmicity of oscillatory firing activity was abolished completely at the steady state. These results suggest that 4-AP-sensitive currents determine the shape and frequency of individual Ca(2+)-dependent action potentials as well as maintaining oscillatory firing activity in PCs.