Effects of toluene exposure on ATPase activity in synaptosomal membranes of rat brain, "in vivo" and "in vitro" studies

Se presenta un trabajo de investigación en el cual se midió la actividad de la enzima Na+K+-ATPasa en la fracción sinaptosomal en cerebro de ratas de la Cepa Wistar expuestas en forma aguda (una exposición) y crónica (30 días consecutivos) a vapores de tolueno, (concentración de 15000 ppm en aire, durante 15 min.) en contraste con un grupo de ratas "control" expuestas al aire libre de tolueno, (estudio in vivo). Los estudios in vitro consistieron en la incubación de la fracción sinaptosomal de cerebro de rata en tubos cerrados conteniendo en el medio reactivo de tolueno a la concentración de 10mM. El cáculo de las constantes de Michaelis-Menten (Km y Vmax) fueron evaluadas en los estudios cinéticos de la enzima en las membranas sinaptosomales. Las ratas tanto de 50 como de 120 días de edad, que fueron expuestas en forma crónica a vapores de tolueno mostraron una reducción de la actividad entre 45 y 56 por ciento con respecto a la actividad del grupo control, siendo estadísticamente significativa (p<0.05). No habiendo diferencias en las ratas expuestas a tolueno en forma aguda. En los experimentos in vitro, también mostraron reducción en la actividad de la enzima podría ser un indicador de los efectos acumulativos de las propiedades físico-químicas del tolueno sobre las membranas biológicas

Efecto del tolueno en el desarrollo neuronal de crías de rata, cuyas madres fueron expuestas a inhalación crónica durante gestación o durante su adolescencia / Effects of the neural development of rat pups whose moters were exposed to toluene via chronic inhalation either during pregnancy or adolescence

Desarrollo neuronal en crios de rata adolescentes y gestantes expuestas a inhalación crónica de tolueno. Se sometieron a inhalación de tolueno a tres ratas jóvenes de 70 días de edad y nueve ratas gestantes de 90 días de edad durante 15 minutos por día desde la cruza hasta un día antes de la fecha de parto. Las ratas jóvenes se cruzaron después del período de inhalación con machos que no inhalaron el solvente. Las crias de las hembras tratadas, así como las de un grupo testigo fueron sometidas a iguales condiciones pero sin tolueno. Fueron sacrificadas por perfusión intracardíaca 24 crias de cada grupo, 12 de 14 días y 12 de 21 días de edad. Fue cuantificado el grado de ramificación de los procesos neuronales de las neuronas piramidales de la V capa de la corteza visual impregnadas con el método rápido de Golgi. Se encontró decremento significativo en las ramificaciones en la porción distal de la dendrita principal en los grupos expuestos al solvente comprarados con los grupos testigo y control

Depolarization of synaptosomal membranes: a study of mechanism by which rhodamine 6G measures membrane potential.

The fluorescence intensity of Rhodamine 6G in synaptosomal suspensions has been measured to monitor the membrane potential changes in pre-synaptic nerve terminals. The fluorescence response of the dye was seen to be a function of potential-dependent partitioning of dye molecules between the synaptosomes and the extracellular medium. Binding of dye molecules to the hydrophobic regions of membranes results in the quenching of fluorescence. Upon depolarization of the synaptosomal membrane, the dye molecules are released from the cells. The effect of changing extracellular ionic composition was also studied. The membrane potential increased linearly with log of [K]0. The resting membrane potential in buffer containing 5 mM K+ was calculated to be -60 mV. Raising the extracellular Ca2+ and Mg2+ from 1.2 mM to 10 mM did not change the membrane potential. Ca2+ ionophore A23187, in the presence of Ca2+ was found to depolarize the membranes.

Ontogeny of ATP and ADP hydrolysis by cerebral cortex synaptosomes from rats

In the present study, we examined the ontogeny of ATP and ADP hydrolysis by cerebral cortex symptosomes from rats of various ages (0-, 7-, 14-, 21- and 60 to 90-day-old rats) in order to learn whether hydrolytic activity increases during the period of intense brain grwth, as has been reported for other enzymes involved in neurotransmitter metabolism. the results demonstrate that ATP and ADP hydrolyzing activities increase in parallel from birth until the second postnatal week (about 4-fold), followed by a slight and statistically insignificant increase until the animal reaches adulthood. The maximum increase in nucleotide hidrolysis coincided with mximum brain growth, which may indicate a role for the enzyme in neurotransmission. Furthermore, the parallel development of both activities (ATPase and ADPase) strongly suggest that a single enzyme, an ATP diphosphohydrolase, is involved in ATP and ADP hydrolisis by the synaptosomal fraction

Effects of chronic treatment with high doses of chlorpromazine on ATP and ADP hydrolysis by synaptosomal fractions from the rat caudate nucleus

Several studies have indicated that chlorpromazine and its metabolites affect ATP hydrolysis by brain and liver plasma membranes in vitro. The present report examines whether chronic treatment (12 days) with high doses of chlorpromazine (10 and 40 mg/kg) could affect ATP and ADP hydrolysis by synaptosomal fractions from the rate caudate nucleus. Both doses of chlorpromazine caused significant and paralled decreases (23 to 31%) in the ATP and ADP hydrolysis. The parallelism between the effects of chlorpromazine on ATP and ADP hydrolys suggests the participation of a single enzyme (ATP diphosphohydrolase) in nucleotide hydrolysis

The secretory effect of canatoxina ou rat brain synaptosomes involves a lipoxygenase-mediated pathway

Canatoxin (CNTX), a neurotoxic protein, is known to activate platelet secretion and aggregation in vitro through a lipoxygenase-dependent pathway. This study shows that CNTX also induces time and dose-dependent serotonin secretion from rat brain synaptosomes. The secretory effect induced by 6 micronM CNTX was similar to that elicited by 150 mM KCl. Nordihyderoguaiaretic acid (500 micronM) completely abolished CNTX-induced serotonin release while 150 micronM indomethacin had no effect. These data suggest the involvement of the lipoxygenase pathway in neurotransmitter release elicited by CNTX as occurs in the platelet

Efeito da dopamina sobre a degradaçäo do hormônio liberador das gonadotrofinas em condiçöes de alteraçäo dos hormônios esteroídes: estudo em sinaptossomos hipotalâmicos da rata / Effect of dopamine on the degradation of gonadotrophin releasing hormone, under steroid hormones changes conditions: study in hypothalamic synaptosomes of rats

A regiäo médio-basal do hipotálamo é rica em terminais nervosos que secretam peptídeos hipofisiotróficos. O estudo in vitro dessa regiäo tem trazido inúmeras contribuiçöes para o conhecimento do controle endócrino do organismo. O procedimento mais simples é a utilizaçäo de fragmentos ou fatias hipotalâmicas porém um método que trouxe muita informaçäo para o estudo das secreçöes hipotalâmicas é a utilizaçäo de sinaptossomos, terminais nervosos isolados. O hormônio liberador das gonadotrofinas (LHRH) está largamente associado com a fraçäo sinaptossômica de homogeneizados hipotalâmicos. Neste trabalho fatores que modulam a liberaçäo desse peptídeo de suspensöes incubadas de sinaptossomos do hipotálamo da rata foram estudados, verificando-se que a dopamina em concentraçöes elevadas ativa a degraduaçäo do LHRH em animais previamente injetados com estradiol ou mistura de estradiol e progesterona. Em ratas ovariectomizadas este efeito só foi verificado quando os animais foram injetados com estradiol ou com estradiol seguido de progesterona, näo ocorrendo nas ratas que receberam o veículo ou progesterona sem estradiol. Sugere-se que essa degradaçäo do LHRH na presença da dopamina, que é dependente dos hormônios esteróides, seja um dos mecanismos fisiológicos de controle das gonadotrofinas hipofisárias