{"id":1129,"date":"2018-10-26T12:16:57","date_gmt":"2018-10-26T12:16:57","guid":{"rendered":"https:\/\/www.ull.es\/servicios\/otri\/?post_type=proyectos-nacionales&p=1129"},"modified":"2018-10-26T12:21:42","modified_gmt":"2018-10-26T12:21:42","slug":"hgc","status":"publish","type":"proyectos-nacionales","link":"https:\/\/www.ull.es\/servicios\/otc\/fr\/proyectos\/proyectos-nacionales\/hgc\/","title":{"rendered":"BFU2017-82618-P. H\u00e9t\u00e9rog\u00e9n\u00e9it\u00e9 des v\u00e9sicules s\u00e9cr\u00e9toires dans la cellule chromatinienne"},"content":{"rendered":"

[vc_row][vc_column][vc_tta_accordion shape=\u00bbsquare\u00bb c_icon=\u00bbchevron\u00bb c_position=\u00bbright\u00bb active_section=\u00bb\u00bb no_fill=\u00bbtrue\u00bb collapsible_all=\u00bbtrue\u00bb][vc_tta_section title=\u00bbResumen\u00bb tab_id=\u00bbresumen\u00bb][vc_column_text]El proceso de la neurotransmisi\u00f3n est\u00e1 basado en la capacidad de las ves\u00edculas secretoras de almacenar neurotransmisores, y en liberar adecuadamente su contenido en respuesta a est\u00edmulos secretores por un proceso universal denominado exocitosis. Es pues el funcionamiento de la ves\u00edcula secretora la clave para el buen funcionamiento de las respuestas fisiol\u00f3gicas y, puede probablemente estar tras la g\u00e9nesis de enfermedades neurol\u00f3gicas y psiqui\u00e1tricas. Este proyecto, est\u00e1 planteado para 3 a\u00f1os y es continuaci\u00f3n de nuestra l\u00ednea de investigaci\u00f3n de los \u00faltimos 25 a\u00f1os. Nos planteamos tres preguntas derivadas de la naturaleza de las ves\u00edculas secretoras y hemos dise\u00f1ado los abordajes experimentales
\npara contestarlas:<\/p>\n

    \n
  1. \u00bfCu\u00e1l es la causa de la enorme a heterogeneidad de las ves\u00edculas secretoras?<\/li>\n
  2. \u00bfQu\u00e9 funci\u00f3n tiene el 70-80% de las ves\u00edculas secretoras, las que no se van a liberar?<\/li>\n
  3. Los problemas funcionales de las ves\u00edculas secretoras, \u00bfPueden estar detr\u00e1s de
    \nlas g\u00e9nesis de la enfermedad de Parkinson?<\/li>\n<\/ol>\n

    Para contestar a las dos primeras preguntas utilizaremos t\u00e9cnicas bien implementadas en nuestro laboratorio: cultivos celulares de cromafines y de feocromocitoma PC12, fraccionamiento subcelular, purificaci\u00f3n de ves\u00edculas secretoras (o gr\u00e1nulos cromafines), DLS (Dynamic Light Scattering), citometr\u00eda de flujo, amperometr\u00eda, electroqu\u00edmica intracelular, HPLC, microscop\u00eda de onda evanescente (TIRFM) y confocal. Tambi\u00e9n la elaboraci\u00f3n de sondas fluorescente mediante t\u00e9cnicas de biolog\u00eda molecular y de prote\u00ednas. Para contestar a la tercera pregunta estudiaremos la capacidad funcional de plaquetas y mastocitos humanos (de voluntarios y de enfermos de Parkinson) para acumular y liberar serotonina ya que estas c\u00e9lulas utilizan mecanismos similares (si no id\u00e9nticos) a los de las neuronas dopamin\u00e9rgicas. Nuestras hip\u00f3tesis de trabajo son que la heterogeneidad de las ves\u00edculas secretoras se debe a una heterogeneidad funcional, que el gran compartimento de ves\u00edculas no liberables funciona como un reservorio m\u00f3vil de catecolaminas para el relleno de los gr\u00e1nulos competentes. Tambi\u00e9n que la muerte selectiva de neuronas dopamin\u00e9rgicas se debe a la acumulaci\u00f3n citos\u00f3lica de dopamina y de DOPAC por el mal funcionamiento de los sistemas de acumulaci\u00f3n vesicular y que este fallo ser\u00e1 com\u00fan (aunque sin esas consecuencias letales) a otras c\u00e9lulas secretoras perif\u00e9ricas que s\u00ed podemos estudiar directamente.
    \n[\/vc_column_text][\/vc_tta_section][vc_tta_section title=\u00bbAbstract\u00bb tab_id=\u00bbabstract\u00bb][vc_column_text]<\/p>\n

    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:<\/p>\n

      \n
    1. What is the cause of the enormous heterogeneity of the secretory vesicles?<\/li>\n
    2. What is the role of 70-80% of secretory vesicles, those that cannot be released?<\/li>\n
    3. The genesis of Parkinson’s disease. Can be caused by functional problems of secretory vesicles?<\/li>\n<\/ol>\n

      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.<\/p>\n

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