Implementación de un cultivo neuronal primario como modelo para el estudio de mecanismos de modulación sobre la vía de señalización de los fosfoinositoles

dc.contributorGómez Correa, María del Pilar
dc.contributorNasi Lignarolo, Enrico
dc.contributorBiofísica de la Señalización Celular
dc.creatorMantilla Esparza, Fabián Andrés
dc.date.accessioned2022-03-02T20:09:50Z
dc.date.available2022-03-02T20:09:50Z
dc.date.created2022-03-02T20:09:50Z
dc.date.issued2021
dc.identifierhttps://repositorio.unal.edu.co/handle/unal/81116
dc.identifierUniversidad Nacional de Colombia
dc.identifierRepositorio Institucional Universidad Nacional de Colombia
dc.identifierhttps://repositorio.unal.edu.co/
dc.description.abstractEl litio se ha usado por décadas como estabilizador del estado de ánimo en trastornos bipolares, pero sus mecanismos de acción a nivel celular aun no se han esclarecido. Aproximaciones bioquímicas resaltan la importancia, entre otras, de la vía de los fosfoinositoles. Nuestro laboratorio reportó efectos potenciadores del litio sobre la movilización de calcio intracelular y corrientes de membrana evocadas por activación de la vía de la fosfolipasa C (PLC) en líneas celulares: el sitio de acción fue acotado a la interacción entre proteína Gq y PLC. La existencia de variadas isoformas de estas proteínas en el sistema nervioso hace pertinente estudiar la generalidad de estos hallazgos en neuronas primarias. En el presente trabajo se implementó un cultivo de neuronas de cerebelo, ya que las neuronas de Purkinje expresan masivamente receptores de IP3 y distintas isoformas de PLC. Por razones de costo y fácil acceso, se utilizaron embriones de pollo. La selección de células candidatas se realizó mediante criterios morfológicos complementados por registros de ‘whole-cell voltage clamp’ que mostraron corrientes capacitivas lentas y de gran magnitud. Su viabilidad fue corroborada por la presencia de corrientes tanto voltaje-dependientes como activadas por quisquilato, un agonista de receptores glutamatérgicos acoplados a Gq/PLC. El quisquilato indujo además incrementos de calcio citosólico, provenientes en parte de reservorios intracelulares y sensibles a inhibición de la PLC. Finalmente, la exposición aguda a litio indujo reducción o potenciación de la respuesta de calcio en distintas células, sugiriendo un efecto diferencial del litio sobre variantes moleculares de PLC y/o Gq. (Texto tomado de la fuente).
dc.description.abstractLithium has been used for decades as a mood stabilizer in bipolar disorder, but its mechanisms of action at cellular level have not been clarified yet. Biochemical approaches highlight the importance, among others, of the phosphoinositides pathway. Our laboratory reported potentiating effects of lithium over the intracellular calcium mobilization and membrane currents evoked by the activation of the Phospholipase C (PLC) pathway in cell lines: the site of action was narrowed down to the interaction between the Gq protein and the PLC. The existence of diverse isoforms of these proteins in the nervous system makes pertinent to study the generality of these findings in primary neurons. In the present work a cerebellar neuronal culture was implemented, given that Purkinje neurons massively express IP3 receptors and diverse isoforms of PLC. Chick embryos were used for reasons of cost and easy access. The selection of candidate neurons was made by morphological criteria complemented by whole-cell voltage clamp recordings that showed slow capacitive currents with big amplitude. Cell viability was corroborated by the presence voltage-dependent currents as well as currents activated by quisqualate, an agonist of glutamatergic receptors coupled to Gq/PLC. Quisqualate induced cytosolic calcium increments, originated in part from intracellular reservoirs, and sensitive to PLC inhibition. Finally, exposure to acute lithium induced reduction or potentiation of the calcium response in different cells, suggesting a differential effect of lithium over molecular variants of PLC and/or Gq.
dc.languagespa
dc.publisherUniversidad Nacional de Colombia
dc.publisherBogotá - Ciencias - Maestría en Ciencias - Biología
dc.publisherDepartamento de Biología
dc.publisherFacultad de Ciencias
dc.publisherBogotá, Colombia
dc.publisherUniversidad Nacional de Colombia - Sede Bogotá
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dc.rightsAtribución-NoComercial-SinDerivadas 4.0 Internacional
dc.rightshttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.rightsinfo:eu-repo/semantics/openAccess
dc.titleImplementación de un cultivo neuronal primario como modelo para el estudio de mecanismos de modulación sobre la vía de señalización de los fosfoinositoles
dc.typeTrabajo de grado - Maestría


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