[vc_row][vc_column][vc_tta_accordion shape=»square» c_icon=»chevron» c_position=»right» active_section=»» no_fill=»true» collapsible_all=»true»][vc_tta_section title=»Resumen» tab_id=»resumen»][vc_column_text]El proceso de la neurotransmisión está basado en la capacidad de las vesículas secretoras de almacenar neurotransmisores, y en liberar adecuadamente su contenido en respuesta a estímulos secretores por un proceso universal denominado exocitosis. Es pues el funcionamiento de la vesícula secretora la clave para el buen funcionamiento de las respuestas fisiológicas y, puede probablemente estar tras la génesis de enfermedades neurológicas y psiquiátricas. Este proyecto, está planteado para 3 años y es continuación de nuestra línea de investigación de los últimos 25 años. Nos planteamos tres preguntas derivadas de la naturaleza de las vesículas secretoras y hemos diseñado los abordajes experimentales to answer them:
To answer the first two questions, we will use techniques well-established in our laboratory: chromaffin and PC12 pheochromocytoma cell cultures, subcellular fractionation, purification of secretory vesicles (or chromaffin granules), dynamic light scattering (DLS), flow cytometry, amperometry, intracellular electrochemistry, HPLC, and transient infrared spectroscopy (TIRFM) and confocal microscopy. We will also develop fluorescent probes using molecular biology and protein techniques. To answer the third question, we will study the functional capacity of human platelets and mast cells (from volunteers and Parkinson's patients) to accumulate and release serotonin, since these cells use mechanisms similar (if not identical) to those of dopaminergic neurons. Our working hypotheses are that the heterogeneity of secretory vesicles is due to functional heterogeneity, that the large compartment of non-releasable vesicles functions as a mobile reservoir of catecholamines for filling competent granules, and that the selective death of dopaminergic neurons is due to the cytosolic accumulation of dopamine and DOPAC resulting from the malfunction of vesicular storage systems. We also hypothesize that this failure is common (albeit without these lethal consequences) to other peripheral secretory cells that we can study directly. [/vc_column_text][/vc_tta_section][vc_tta_section title=»Abstract» tab_id=»abstract»][vc_column_text]
The process of neurotransmission is based on the ability of the secretory vesicles to store neurotransmitters and on the ability to adequately release their content, in response to secretory stimuli, by a universal process called exocytosis. It is therefore the function of the secretory vesicle the key of the physiological responses and, probably, be behind the genesis of neurological and psychiatric diseases. This project is planned for 3 years and is a continuation of our line of research for the last 25 years. We posed three questions derived from the nature of the secretory vesicles and we have designed the experimental approaches to answer them:
To answer the first two questions, we will use well-implemented techniques in our laboratory: cell cultures of chromaffin cells and pheochromocytoma PC12, subcellular fractionation, secretory vesicle purification (or chromaffin granules), DLS (Dynamic Light Scattering), flow cytometry, amperometry, Intracellular electrochemistry, HPLC, evanescent wave (TIRFM) and confocal microscopy. Also, we will develop fluorescent probes using techniques of molecular biology and proteins. To answer the third question we will study the functional capacity of human platelets and mast cells (from volunteers and Parkinson's patients) to accumulate and release serotonin as these cells use similar (if not identical) mechanisms to those of dopaminergic neurons. Our working hypotheses is that the heterogeneity of the secretory vesicles is due to functional heterogeneity, that the large compartment of non-releasable vesicles functions as a mobile reservoir of catecholamines for the filling of the competent granules. Also that the selective death of dopaminergic neurons is due to the cytosolic accumulation of dopamine and DOPAC due to the malfunctioning of vesicular accumulation systems and that this failure will be common (although without those lethal consequences) to other secretory cells that we can be directly studied.
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