Altered oligodendrocytes in spinal enlargements of streptozotocin diabetic rats / Oligodendrocitos alterados en la intumescencia espinal de ratas diabéticas con estreptozotocina

SUMMARY: Disturbances of sensory and motor nerve conduction velocity in the spinal cord as well as degenerated myelin sheaths are observed in diabetic patients and animal models. Indeed, oligodendrocytes (OLs), which are important neuroglial cells, generate myelin in the central nervous system. Spinal enlargement, including cervical and lumbar enlargements, innervates all limbs. Thus, the purposes of this study were to examine and compare the ultrastructural alterations of OLs in spinal enlargements of streptozotocin (STZ)- induced diabetic rats and controls. Thirteen male Sprague-Dawley rats were induced with STZ in citrate buffer and six control rats were injected with the same buffer solution. All rats were sacrificed after inductions at four (short-term DM) and twenty-four weeks (long-term DM). The selected spinal enlargements were processed for transmission electron microscopy. The OL alterations in both the cervical and lumbar enlargements were apparently the same. In short-term DM, the nuclei of OLs became swelled with chromatin clumping. Cytoplasmic organelles were moderately damaged. In long-term DM, OLs contained shrinkage nuclei with thick heterochromatin clumping. Severely degenerated mitochondria with disrupted cristae and broken membranes were observed. Moreover, distended and fragmented rough endoplasmic reticulum were observed, and large clear areas were present in the cytoplasm. Additionally, the loosening, splitting, and destruction of myelin lamellae were found. This study can provide important preliminary information about the alteration of OLs in the spinal cords of diabetic patients, which might be involve in the impairments of sensory and motor conduction velocities in these individuals.

RESUMEN: En pacientes diabéticos y modelos animales se observan alteraciones de la velocidad de conducción nerviosa sensorial y motora en la médula espinal, así como vainas de mielina degeneradas. De hecho, los oligodendrocitos (OL), que son importantes células neurogliales, generan mielina en el sistema nervioso central. La intumescencia espinal, a nivel cervical y lumbar, inerva los miembros. Por lo tanto, los propósitos de este estudio fueron examinar y comparar las alteraciones ultraestructurales de los OL en la intumescencia espinal de ratas diabéticas inducidas por estreptozotocina (STZ) y controles. Se indujeron trece ratas macho Sprague-Dawley con STZ en tampón citrato y se inyectaron seis ratas de control con la misma solución tampón. Todas las ratas se sacrificaron después de la inducción a las cuatro (DM a corto plazo) y a las veinticuatro semanas (DM a largo plazo). Las ampliaciones de la columna seleccionadas se procesaron para microscopía electrónica de transmisión. Las alteraciones de OL en las intumescencias cervical y lumbar eran aparentemente las mismas. En la DM a corto plazo, los núcleos de los OL se hincharon con la acumulación de cromatina. Los orgánulos citoplasmáticos sufrieron daños moderados. En la DM a largo plazo, los OL contenían núcleos de contracción con aglutinación de heterocromatina gruesa. Se observaron mitocondrias severamente degeneradas con crestas y membranas rotas. Además, se observó un retículo endoplásmico rugoso distendido y fragmentado, y estaban presentes grandes áreas claras en el citoplasma. Además, se encontraron el aflojamiento, la división y la destrucción de las laminillas de mielina. Este estudio puede proporcionar información preliminar importante sobre la alteración de los OL en la médula espinal de los pacientes diabéticos, que podría estar involucrada en las alteraciones de las velocidades de conducción sensorial y motora en estos individuos.

Does neuroectodermal stem cells transplantation restore neural regeneration and locomotor functions in compressed spinal cord injury rat model? / ¿El trasplante de células madre neuroectodérmicas restaura la regeneración neural y las funciones locomotoras en el modelo de rata con lesión de la médula espinal comprimida?

SUMMARY: The aim of this study was to determine the possible regenerative effect of neuroectodermal stem cells on the ultrastructural, and locomotor function resulting from compressed injury to the spinal cord in a rat model. Forty male rats were divided into control and sham groups (20 rats each). Compressed spinal cord injured (CSCI) were forty rats which subdivided equally into: untreated, treated by neuroectodermal stem cells (NESCs). After four weeks, all rats in different groups were scarified, samples were taken from central, cranial, and caudal to the site of spinal cord injury. Specimens were prepared for light and electron microscopic examination. The number of remyelinated axons in central, cranial and caudal regions to the injured spinal cord after transplantation of NESCs was counted. The open field test assessed the locomotor function. Results revealed that compressed spinal cord injury resulted in loss and degeneration of numerous nerve fibers, myelin splitting and degeneration of mitochondria. Four weeks after transplantation of NESCs regenerated axons were noticed in cranial and central sites, while degenerate axons were noticed caudal to the lesion. Number of remyelinated axons was significantly increased in both central and cranial to the site of spinal cord injury in comparison with caudal region which had the least number of remyelinated axons. Transplantation of NESCs improved significantly the locomotor functional activity In conclusion, neuroectodermal stem cells transplantation ameliorated the histopathological and ultrastructural changes, and improved the functional locomotor activity in CSCI rat.

RESUMEN: El objetivo de este estudio fue determinar el posible efecto regenerativo de las células madre neuroectodérmicas en la función ultraestructural y locomotora de una lesión comprimida en la médula espinal en un modelo de rata. Cuarenta ratas macho se dividieron en grupos control y sham (20 ratas en cada grupo). La médula espinal lesionada (CSCI) tenía cuarenta ratas que se subdividieron de igual forma en los siguientes grupos: no tratadas, tratadas con células madre neuroectodérmicas (NESCs). Al término de cuatro semanas, todas las ratas en los diferentes grupos fueron escarificadas, se tomaron muestras de las áreas central, craneal y caudal en relación al sitio de la lesión de la médula espinal. Las muestras fueron preparadas para examen microscópico de luz y electrónica. Se contó el número de axones remielinizados en las regiones central, craneal y caudal de la médula espinal lesionada después del trasplante de NESCs. La prueba de campo abierto evaluó la función locomotora. Los resultados revelaron que la lesión de la médula espinal comprimida provocó la pérdida y degeneración de numerosas fibras nerviosas, la división de la mielina y la degeneración de las mitocondrias. Cuatro semanas después del trasplante de NESCs, se notaron axones regenerados en los sitios craneales y centrales, mientras que los axones degenerados se notaron caudal a la lesión. El número de axones remielinizados aumentó significativamente tanto en el centro como en el cráneo hasta el sitio de la lesión de la médula espinal en comparación con la región caudal que tenía el menor número de axones remielinizados. El trasplante de NESCs mejoró significativamente la actividad funcional locomotora. En conclusión, el trasplante de células madre neuroectodérmicas mejoró los cambios histopatológicos y ultraestructurales, y mejoró la actividad locomotora funcional en la rata CSCI.

Efeitos neurotóxicos de sulfato de magnésio intratecal / Neurotoxic effects of intrathecal magnesium sulphate / Efectos neurotóxicos de sulfuro de magnesio intratecal

JUSTIFICATIVA E OBJETIVOS: Avaliar os potenciais efeitos neurotóxicos em nível ultraestrutural desulfato de magnésio administrado por via intratecal em dose única ou múltipla. MÉTODOS: Estudo realizado com 24 ratos Sprague-Dawley, peso médio entre 250 e 300 g. Apósjejum de 4 horas, os ratos receberam 10 mg.kg-1 de cloreto de xilazina por via intraperitoneale, em seguida, foram divididos aleatoriamente em três grupos. Grupo I (n = 8) recebeu 0,9% desoro fisiológico normal, Grupo II (n = 8) recebeu uma injeção de 0,02 mL de sulfato de magnésioa 15% por via intratecal e Grupo III (n = 8) recebeu 0,02 mL de sulfato de magnésio a 15% umavez por dia durante sete dias. As injeções foram aplicadas dentro de 0,40x50 milímetros daárea lombar. Após sete dias, os animais foram sacrificados sob anestesia com uma injeção deformaldeído a 10% na aorta e os tecidos foram fixados. A medula espinal foi, então, examinadae histopatologicamente avaliada sob microscópio eletrônico. O teste de Kruskal-Wallis foi usadopara avaliação estatística. Um valor de p < 0,05 foi considerado estatisticamente significativo. RESULTADOS: Neurodegeneração significativa foi detectada nos ratos que receberam uma únicainjeção ou injeções repetidas de sulfato de magnésio, em comparação com o grupo controle. O escore na avaliação histopatológica desse grupo também foi alto. CONCLUSÃO: Com base no exame de microscopia eletrônica, descobrimos que a administraçãointratecal de sulfato de magnésio induziu neurodegeneração.

BACKGROUND AND OBJECTIVES: To assess the potential neurotoxic effects at the ultrastructural level of magnesium sulfate administered intrathecally as a single or multi-dose. METHODS: Our study was conducted with 24 Sprague-Dawley rats that weighed 250-300 g. After a 4-hour fast, the rats were given 10 mg.kg-1 xylazine chloride intraperitoneal and then randomly allocated into three groups. Group I (n = 8) received 0.9% normal saline, Group II (n = 8) was given one intrathecal injection of 0.02 mL of 15% magnesium sulphate, and Group III (n = 8) was given 0.02 mL of 15% magnesium sulphate once a day for seven days. The injections were given within 0.40x50 mm from the lumbar area. After seven days, the animals were sacrificed under anesthesia with an aortic injection of 10% formaldehyde and their tissues were fixed. The medulla spinalis was then examined and histopathologically evaluated under an electron microscope. The Kruskal-Wallis test was used for statistical evaluation. A value of p < .05 was considered to be statistically significant. RESULTS: Significant neurodegeneration was detected in rats given single or repeated magnesium sulphate injections compared to the control group. The histopathological evaluation score of this group was also high. CONCLUSIONS: Based on electron microscopic examination, we found that intrathecal magnesium sulphate administration induced neurodegeneration.

JUSTIFICATIVA Y OBJETIVOS: Evaluar los potenciales efectos neurotóxicos en nivel ultraestructural de sulfuro de magnesio administrado por vía intratecal en dosis única o múltiple. MÉTODOS: Estudio realizado con 24 ratones Spraque-Dawley, con un peso promedio entre los 250 y los 300 g. Después del ayuno de 4 horas, los ratones recibieron 10 mg.kg-1 de cloruro de xilazina por vía intraperitoneal y enseguida fueron divididos aleatoriamente en tres grupos. El grupo I (n = 8) recibió 0,9% de suero fisiológico normal, Grupo II (n = 8) recibió una inyección de 0,02 mL de sulfuro de magnesio al 15% por vía intratecal y Grupo III (n = 8) recibió 0,02 mL de sulfuro de magnesio al 15% una vez por día durante siete días. Las inyecciones fueron aplicadas dentro de 0,40x50 milímetros del área lumbar. Después de siete días, los animales fueron sacrificados con anestesia con una inyección de formaldehido al 10% en la aorta y los tejidos fueron pegados. La médula espinal se examinó y fue histopatológicamente evaluada bajo microscopio electrónico. El test de Kruskal-Wallis fue usado para la evaluación estadística. Un valor de p < 0,05 fue considerado estadísticamente significativo. RESULTADOS: La neurodegeneración significativa fue detectada en los ratones que recibieron una sola inyección o inyecciones repetidas de sulfuro de magnesio, en comparación con el grupo control. El puntaje en la evaluación histopatológica de ese grupo también fue alto. CONCLUSIONES: Basándonos en el examen de microscopía electrónica, descubrimos que la administración intratecal de sulfuro de magnesio indujo a la neurodegeneración.

The immunomodulator glatiramer acetate influences spinal motoneuron plasticity during the course of multiple sclerosis in an animal model

The immunomodulador glatiramer acetate (GA) has been shown to significantly reduce the severity of symptoms during the course of multiple sclerosis and in its animal model - experimental autoimmune encephalomyelitis (EAE). Since GA may influence the response of non-neuronal cells in the spinal cord, it is possible that, to some extent, this drug affects the synaptic changes induced during the exacerbation of EAE. In the present study, we investigated whether GA has a positive influence on the loss of inputs to the motoneurons during the course of EAE in rats. Lewis rats were subjected to EAE associated with GA or placebo treatment. The animals were sacrificed after 15 days of treatment and the spinal cords processed for immunohistochemical analysis and transmission electron microscopy. A correlation between the synaptic changes and glial activation was obtained by performing labeling of synaptophysin and glial fibrillary acidic protein using immunohistochemical analysis. Ultrastructural analysis of the terminals apposed to alpha motoneurons was also performed by electron transmission microscopy. Interestingly, although the GA treatment preserved synaptophysin labeling, it did not significantly reduce the glial reaction, indicating that inflammatory activity was still present. Also, ultrastructural analysis showed that GA treatment significantly prevented retraction of both F and S type terminals compared to placebo. The present results indicate that the immunomodulator GA has an influence on the stability of nerve terminals in the spinal cord, which in turn may contribute to its neuroprotective effects during the course of multiple sclerosis.

protective role of ginger [zingiber officinale] against acrylamide induced neurotoxicity in mice

To evaluate the possible improvement in the histological and ultrastructural features of spinal cord and cerebellum of mice treated with ginger together with acrylamide. Thirty male adult CD-1 mice were divided into three groups, the first group [10 mice, served as control, the second group[10 mice] received 200 p.p.mn of acrylamide for 10 weeks [3 days/week] and the third group [10 mice] animals received ethanolic ginger extract at 50 mg/L [-5]for 10 weeks [3 days/week]. The animals were sacrificed and us sue samples were taken and processed for ultrastructural study. Acrylamide administration induced necrosis of the spinal cord motor neurons, degeneration of the myelinated and non-myelinated nerve fibers. The neuropil exhibited edema, vaculation and shrinkage of cerebellum neurons. In addition the damage of most cellular organdies such mitochondria, Golgi complex, rough endoplasmic reticulum, and irregularity of the nuclei which became peripheral in position, edema of tile neuropil and damage of the synaptic sites. On the contrary, in animals received ginger extract, the nervous tissue revealed improvement of the histology and ultrastructure of the tissues which became almost similar to tile control group tissue. It is possible to suggest that ginger may have a significant importance in protection against acrylamide induced neurological damage

Effect of acrylamide on the central nervous system of rates: histological, histochemical, and electron microscopy studies

The effect of acrylamide on the central nervous system of rats was studied in order to determine its neurotoxicity effect. Seventy rats were used in the present study which were divided into three groups. The rats of group one were used as control while rats of group two were given acrylamide by intramuscular injection with a dose of 5 mg/kg body weight, and the rats of group three were given 25 mg/kg acrylamide twice weekly for one month. The rats were sacrificed after 1, 2, 3, and 4 weeks of acrylamide injection and cervical spinal cord was removed and processed for histological, histochemical, and electron microscopical studies. Histological results showed mild degenerative changes of nerve fibers after one and two weeks of acrylamide injection while after three and four weeks severe changes were seen following both doses of acrylamide. Histochemical changes appeared in the decreased activities of cytochrome oxidase, RNA, and Nissl substances after one and two weeks of acrylamide injection and this decrease reached its minimal activity at the end of the experiment for the two doses while acetyl-cholinesterase and DNA were found to be increased. Electron microscopic study revealed irregular outline of nuclear envelop with some nuclear pores, vascular degeneration of cytoplasmic organelles, demyleinated and thinly myleinated axons, and many hypertrophied mitochondria. All these results indicated that acrylamide which is still used in the synthesis of polymers for a variety of industrial applications, is highly neurotoxic

Desmielinizaçäo tóxica do sistema nervoso central: I. Açäo de uma droga intercalante gliotóxica na medula espinhal de ratos Wistar / Toxic demyelination of the central nervous system: I. Effects of an intercalating gliotoxic chemical in the spinal cord of Wistar rats

Pequenos volumes de brometo de etídio foram injetados nas colunas dorsais da medula lombar de ratos Wistar. Foi assim induzido processo desmielinizante que variou em natureza e velocidade de reparaçäo de acordo com a dose empregada. As lesöes produzidas foram classificadas em três tipos (I, rápidas: II, lentas; III, intermediárias), de acordo com as características histológicas e a extensäo da remielinizaçäo. Em algumas lesöes ou em áreas dentro da mesma lesäo, os restos de mielina e de células eram rapidamente processados por macrófagos e os axônios rapidamente remielinizados por células de Schwann, em quanto em outras lesöes de duraçäo similar, ou em áreas dentro da mesma lesäo, a mielina se transformava em emaranhados de membranas que persistiam ao redor do axônio por longos períodos de tempo. Nas lesöes que continham tais membranas derivadas da mielina, os macrófagos eram escassos e a remielinizaçäo, feita pelas células de Schwann, era demorada e trabalhosa. Concluiu-se que a resoluçäo lenta de algumas lesöes resultara do lapso transcorrido entre a intoxicaçäo e o desaparecimento das células relacionada a mielina, significando que as respostas celulares a desmielinizaçäo tiveram lugar em área livre de células gliais. Esta näo podia sustentar, portnto, a movimentaçäo celular necessária para a remoçäo dos restos de mielina e a posterior remielinizaçäo. Esta investigaçäo indica que o desenvolvimento e o desfecho da desmielinizaçäo podem ser alterados pelos eventos celulares que acompanham a degeneraçäo dos oligodendrócitos

Desmielinizaçäo tóxica do sistema nervoso central: II. Aspectos biológicos das células de Schwann observados durante o processo de reparaçäo do tecido / Toxic demyelination of the central nervous system: II. Biological aspects of Schwann cells observed during the process of tissue repair

A droga gliotóxica brometo de etídio, quando injetada localmente na medula espinhal do rato, induziu áreas de desmielinizaçäo que se desenvolveram em tempos diferentes de acordo com a dose empregada. Doses altas induziram lesöes de desenvolvimento rápido (tipo I) e intermediárias (tipo III), enquanto doses baixas induziram lesöes lentas (tipo II). Após o processo desmielinizante, os azônios desprovidos de suas bainhas de mielina foram remielinizados por oligodendrócitos ou células de Schwann (CS) dependendo de sua localizaçäo em áreas contendo ou näo astrócitos, respectivamente. Na maioria das lesöes, a área remielinizada pelas CS era proeminente. Nas lesöes que repararam mais lentamente foi possível observar os fatores que influenciaram o comportamento dessas células dentro dos limites do sistema nervoso central. Após a invasäo inicial a partir da superfície pial e dos espaços preivasculares, a expansäo das CS dependeu da presenca de matriz extracelular estável. nas lesöes de tipo I, esta matriz estava presente devido a natureza inflamatória do processo. Nas lesöes de tipo II a matriz näo ocorreu e as CS só podiam migrar entre os axônios desmielizados usando-os tal como uma passadeira. Entre as células contíguas podiam ser observadas fibras colágenas de diâmetro pequeno. Näo foi evidenciada migraçäo das CS ao longo dos axônios

On the Origin of Oligodendrocytes

Development and differentiation of astrocytes and oligodendrocytes (OC) in the developing human fetal spinal cord (HFSC) have been investigated by the correlative analysis of light microscopic, EM, Golgi and immunocytochemical studies. The evidence is presented to suggest, (a) that radial glia are the first distinguishable neuroglial element among the cells within the ventricular zone, (b) that radial glia contains astrocyte-specific glial fibrillary acidic protein (GFAP), (c) that radial glia undergoes transformation into astroglial cells, (d) that "transitional forms" possessing the light and EM features of both astroglial and oligodendroglial cells appear just prior to the onset of myelination, and (e) the myelin-forming OC are most likely derived from radial glial cells, either directly or through intermediated astroglial forms.