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Caracterización funcional del canal de potasio activado por calcio de conductancia intermedia (KCa3.1) en el endotelio de la córnea en condiciones fisiológicas y en ambientes hiperglúcidos

dc.contributor.advisorMatheus Merino, Luisa Marina
dc.contributor.gruplacGrupo de Neurociencias de la Universidad del Rosario (NEUROS)es
dc.creatorAmador Muñoz, Diana Patricia
dc.creator.degreeDoctor en Ciencias Biomédicas y Biológicases
dc.creator.degreeLevelDoctorado
dc.creator.degreetypeFull timees
dc.date.accessioned2021-10-09T15:07:07Z
dc.date.available2021-10-09T15:07:07Z
dc.date.created2021-06-22
dc.date.embargoEndinfo:eu-repo/date/embargoEnd/2025-10-11
dc.descriptionLa córnea es el lente que protege la superficie anterior del ojo y su transparencia es clave para permitir la visión. Esta característica en gran medida está determinada por la actividad de las células de su capa más profunda, el endotelio corneal. Una monocapa de células hexagonales cuyas características morfo-fisiológicas le permiten no solo compensar la tendencia a la sobrehidratación natural que tienen capas más superficiales de la córnea, especialmente el estroma, sino ser un punto clave para el ingreso y la distribución de nutrientes a nivel corneal. Dado que tras el nacimiento el potencial proliferativo de las células endoteliales de la córnea humana es extremadamente limitado, la densidad celular se reduce progresivamente durante la vida, y para restaurar el tejido, las células adyacentes deben migrar y cubrir el área que se ha dañado. Las patologías que afecten, directa o indirectamente, al endotelio corneal aceleran la perdida celular y generan una disfunción que, en última instancia, conlleva a la pérdida de la transparencia corneal haciendo casi imposible la visión, y en la actualidad el único tratamiento disponible es el trasplante. En las últimas décadas, la diabetes mellitus (DM) se ha identificado como una de las enfermedades sistémicas que afectan el endotelio corneal. En estos pacientes se ha descrito un aumento de la paquimetría, una reducción del recuento de células endoteliales respecto a personas sanas de la misma edad y sexo, e incluso diferencias entre pacientes diabéticos de acuerdo con el tiempo de evolución de la enfermedad, además, tras procedimientos quirúrgicos, la alteración de la función de esta barrera ocular y el edema del estroma suelen ser persistentes. Sin embargo, los mecanismos fisiopatológicos por los que la DM afecta el endotelio de la córnea están pobremente descritos. La DM corresponde a un grupo de trastornos metabólicos cuya condición sine qua non es la hiperglicemia. Si bien no es la única causa de hiperglicemia en el ser humano, sí es la que se relaciona con un aumento persistente de la concentración de glucosa en la sangre y otros líquidos extracelulares. Esta situación lleva a complicaciones que afectan preferencialmente células que, como el endotelio corneal, internalizan la glucosa por medio de transportadores de glucosa tipo 1 (GLUT1), es decir por transportadores independientes de los niveles de insulina en sangre, así como ocurre en los eritrocitos, los astrocitos, las neuronas y las células renales, células con las que los tejidos oculares muestran cierta homología desde su origen embrionario (el endotelio deriva de la cresta neural), hasta la fisiopatología, puesto que clínicamente existe concordancia entre el compromiso renal y el ocular. La exposición persistente de las células de todos los tejidos a niveles elevados de glucosa induce lesiones que, en general, están relacionadas con un desbalance en el que la glucosa, y otros metabolitos, se convierten en sustrato de vías metabólicas que usualmente no los utilizan y favorecen el desarrollo de alteraciones morfológicas y funcionales, que una vez se desarrollan son prácticamente irreversibles. Sin embargo, aunque para los tejidos nervioso, cardiovascular y renal la lesión mediada por un microambiente diabético está bien caracterizada, no sucede lo mismo para la córnea y menos aún para el endotelio corneal. Los estudios en diversas poblaciones que han intentado evaluar el impacto de la enfermedad sobre el endotelio, aunque son consistentes en cuanto a los cambios morfológicos, reportan resultados discordantes en cuanto al recuento endotelial y la paquimetría. Por lo anterior, se consideró relevante evaluar el impacto de la diabetes sobre el endotelio mediante modelos estadísticos que permitieran discriminar los cambios asociados a la edad descritos para estas células; en particular, para este análisis era importante identificar el efecto de la enfermedad sobre la densidad celular del endotelio y su capacidad para mantener la deshidratación relativa del estroma y controlar el espesor corneal. Así que se construyó un modelo estadístico con base en una meta-regresión que incluía los tipos de diabetes y la edad como moduladores, para evaluar el impacto real de cada tipo de DM (DM tipo 1 y DM tipo 2) sobre la densidad del endotelio corneal y el espesor de la córnea, determinado por la paquimetría. Este análisis evidenció que el recuento celular se reducía significativamente por la enfermedad, predominantemente en pacientes con DM tipo 1 en quienes el compromiso era independiente de la duración de la enfermedad, y que el aumento del espesor corneal en los pacientes diabéticos era superior al esperado por edad para ambos grupos, independientemente del tipo de diabetes. Estos resultados, que evidenciaban clínicamente el impacto de la hiperglicemia sobre el endotelio, sustentaron la necesidad de evaluar in vitro el efecto que tienen las concentraciones elevadas de glucosa sobre estas células. Particularmente sobre su capacidad de proliferación, su capacidad de migrar para cubrir un defecto y en la inducción de apoptosis, principal tipo de muerte celular identificado hasta el momento en estas células, y el cual ha sido descrito dentro de las respuestas de las células endoteliales frente al estrés oxidativo condición clave dentro de la fisiopatología diabética. Para evaluar los cambios en la capacidad de proliferación de las células del endotelio, se utilizaron cultivos celulares de una línea inmortalizada, las cuales se expusieron al medio definido para ellas como basal y a medios con altas concentraciones de glucosa (55mM). y se hizo seguimiento de la tasa de cambio en la densidad celular mediante ensayos de reducción de MTT [bromuro de 3- (4, 5-dimetiltiazol-2-il) -2,5-difeniltetrazolio]. En estos experimentos, las células con metabolismo activo convierten el MTT en un producto de color púrpura que una vez solubilizado permite evaluar mediante colorimetría los cambios en la cantidad de células viables. Las pruebas realizadas permitieron evidenciar un aumento en la cantidad de células de los cultivos expuestos a altas concentraciones de glucosa (55mM), mostrando una diferencia significativa tras 24 horas; sin embargo, aunque la diferencia permanecía siendo significativa tras 48 horas, a partir de ese momento el recuento celular medido indirectamente por el método colorimétrico mostraba una reducción progresiva que igualaba la cantidad de células viables a los 5 días para las dos condiciones. La influencia de las distintas concentraciones de glucosa en la capacidad del endotelio para cerrar un defecto de continuidad en la monocapa, se realizó utilizando el método descrito por Liang y colaboradores en 2007 (Liang et al., 2007), en el que se crea un rasguño ("scratch") en una monocapa celular, y se hace seguimiento imagenológico mediante fotografías tomadas desde el momento en que se genera la lesión y a intervalos regulares, hasta que la “herida” cierra. Esto permite comparar el tiempo que le toma a las células del endotelio cerrar el espacio de la lesión bajo diferentes condiciones y cuantificar la tasa de migración de las células. En estos experimentos, se evidenció un retraso significativo de las células expuestas a elevadas concentraciones de glucosa para cerrar el defecto en comparación con células en medios basales. Mientras estas últimas tomaban aproximadamente 5-6 días para cerrar el defecto, para ese momento las células expuestas a niveles elevados de glucosa habían cerrado, en promedio, 50% de la distancia. Por último, la evaluación de apoptosis se realizó mediante un kit comercial (Cell Death Detection ELISA PLUS (Roche) ) que utiliza la técnica de ELISA (inmunoensayo enzimático) tipo sándwich con anticuerpos monoclonales para histonas con el fin de determinar mono y oligonucleosomas en la fracción citoplasmática de los lisados celulares de cada condición. Estos experimentos evidenciaron que la exposición a elevadas concentraciones de glucosa por 24 horas inducía apoptosis tres veces superior a la que se presentaba en las células en condiciones basales. En la fisiopatología de los procesos deletéreos asociados a la diabetes recientemente se ha identificado la importancia de los procesos electrofisiológicos. En células renales y de la microglía se ha demostrado que los canales de potasio activados por calcio de conductancia intermedia (KCa3.1) parecen tener un papel relevante. Sin embargo, la familia de canales de potasio activados por calcio (KCa) no había sido descrita previamente en el endotelio corneal por lo que fue necesario inicialmente identificar cuales canales de esta familia se expresaban en estas células. Se partió de un análisis bioinformático que permitiera la identificación in silico de estos canales tras lo cual, se comprobó in vitro mediante PCR y en algunos casos Western blot e inmunomarcación la presencia de los canales de potasio activado por calcio de baja conductancia KCa2.2 y KCa2.3, el de conductancia intermedia KCa3.1 y el canal de potasio activado por sodio KNa2.1 (Slick) en las células del endotelio corneal. En el trascurso del desarrollo del presente trabajo, Anumanthan y colaboradores (2018) publicaron un artículo que evaluaba la actividad de KCa3.1 en el estroma de la córnea que incluyó microfotografías que permitían ver marcado el endotelio, lo que reforzó los resultados obtenidos. Se procedió a estudiar que funciones cumplía KCa3.1 en el endotelio en condiciones basales y si estas se modificaban ante la exposición de las células a concentraciones elevadas de glucosa. Estos experimentos que incluyeron la estimulación e inhibición química del canal permitieron identificar la participación de canales KCa3.1 en los procesos de migración, proliferación y apoptosis. KCa3.1, el canal de conductancia intermedia de la familia de canales de potasio activados por calcio, puede ser activado por 1-1-Etil-1,3-dihidro-2H-benzimidazol-2-ona (benzimidazolona) (EBIO-1) y puede ser inhibido selectivamente por 1-[(2-Clorofenil) difenilmetil]-1H-pirazol (TRAM-34). Estos compuestos se utilizaron para probar el efecto de la estimulación e inhibición del canal en la proliferación, migración y apoptosis de las células del endotelio corneal expuesto tanto a condiciones basales como a condiciones hiperglúcidas. La estimulación del canal con EBIO-1 mostró un efecto inhibitorio significativo sobre la proliferación de las células del endotelio cuando se utiliza a concentraciones de 50, 100 y 200 µM, suficiente para contrarrestar el efecto proliferativo identificado en condiciones de alta glucosa durante los primeros días. La inhibición del canal con TRAM-34 a concentraciones de 2, 4 y 8 µM evidenció un efecto contrario, aumentó la proliferación de las células en condiciones basales, especialmente a las concentraciones más altas, y potenció el efecto proliferativo identificado en condiciones de alta glucosa, manteniendo una cantidad mayor de células viables por un tiempo más prolongado. En cuanto a la migración, la estimulación con EBIO-1 redujo la tasa de migración aproximadamente un 50% en condiciones basales y potenció el efecto visto en las condiciones hiperglúcidas. Por el contrario, la inhibición de KCa3.1 con TRAM-34 a 2 µM, aceleró la migración e incluso acercó la tasa de migración de las células en condiciones de alta glucosa a las de las células basales, los efectos son menores a concentraciones mayores. Finalmente, en condiciones basales ninguna de las concentraciones descritas para EBIO-1 y para TRAM-34 aumentaron la tasa de apoptosis; sin embargo, asociadas a medios con elevadas concentraciones de glucosa, EBIO-1 a concentraciones de 100 µM y TRAM_34 a 4 µM sí lo hicieron. En conclusión, este trabajo permitió identificar el compromiso de la diabetes mellitus sobre el endotelio corneal mediante la determinación de su rol en la reducción de la densidad de esta monocapa más allá de la esperada fisiológicamente por la edad, y su impacto en el aumento de la paquimetría, además de identificar un compromiso más severo de los pacientes con diabetes mellitus tipo 1 respecto a los que cursan con el tipo 2. Adicionalmente, se describió por primera vez la presencia de canales de potasio activados por calcio de conductancia baja, además de confirmar la expresión del canal de conductancia intermedia, y se identificaron en el endotelio corneal los canales de potasio activados por sodio de alta conductancia tipo 2. Por último, se exploró la participación de KCa 3.1 en la proliferación, migración y apoptosis de estas células y se describió su papel como moduladores de estos procesos tanto en condiciones basales como en condiciones de alta glucosa lo cual es relevante tanto en condiciones fisiológicas como en condiciones patológicas, no solo en el escenario de la diabetes sino probablemente en las respuestas ante otros eventos.es
dc.description.abstractThe cornea is the lens that protects the anterior surface of the eye and its transparent nature is a crucial part of the functioning of the eye. This characteristic is mainly determined by the activity of the cells of its deepest layer, the corneal endothelium. This is a monolayer of hexagonal cells whose morpho-physiological characteristics allow it not only to compensate the natural hyperhydration of the superficial layers of the cornea, especially the stroma, but also mean it can be a key point for the entry and distribution of nutrients in the cornea. Since the proliferative potential of human corneal endothelial cells after birth is extremely limited, cell density progressively reduces during life, and to restore the tissue, adjacent cells must spread out and cover the area that has been damaged. Pathologies that directly or indirectly affect the corneal endothelium accelerate cell loss and generate dysfunction that ultimately leads to loss of corneal transparency, severely hampering vision, and the only treatment currently available is a transplant. In recent decades, diabetes mellitus (DM) has been identified as one of the systemic diseases that affect the corneal endothelium. In these patients, the literature has described increases in pachymetry, reductions in the endothelial cell count with respect to healthy people of the same age and sex, and even differences between diabetic patients depending on the time of disease progression. Furthermore, the alteration of the function of this ocular barrier and the stromal edema often persists even after surgical procedures. However, the pathophysiological mechanisms by which DM affects the corneal endothelium are poorly described. DM is one of a group of metabolic disorders whose sine qua non is hyperglycaemia. Although it is not the only cause of hyperglycaemia in humans, it is the one that is related to a persistent increase in glucose levels in the blood and other extracellular liquids. This leads to complications that disproportionally affect cells that, like the corneal endothelium, absorb glucose through glucose transporter 1 (GLUT1), that is to say through transporters that are independent of blood insulin levels, as occurs in erythrocytes, astrocytes, neurons and renal cells, cells which show certain similarity to eye tissue in terms not only of their embryonic origin (the endothelium derives from the neural crest), but also in their physiopathology, since there are clinical similarities between renal and ocular involvement Persistent exposure of cells in all tissues to high levels of glucose induces lesions that, in general, are related to an imbalance in which glucose and other metabolites become substrates for metabolic pathways that usually do not use them, favoring the development of morphological and functional alterations that, once they have occurred, are practically irreversible. However, although lesions related to diabetic microenvironments are well characterized for nervous, cardiovascular and renal tissues, the same is not true for the cornea and even less so for the corneal endothelium. Studies in various populations that have attempted to evaluate the impact of the disease on the endothelium, while consistent in terms of morphological changes, give discordant results in terms of endothelial count and pachymetry. Therefore, we considered it important to evaluate the impact of diabetes on the endothelium by means of statistical models that make it possible to discriminate the age-associated changes described for these cells; in particular, for this analysis it was important to identify the effect of the disease on endothelial cell density and its ability to maintain relative stromal dehydration and control corneal thickness. So, a statistical model was constructed, based on a meta-regression that included type of diabetes and age as modulators, to evaluate the real impact of each type of DM (type 1 DM and type 2 DM) on corneal endothelial density and corneal thickness, as determined by pachymetry. This analysis showed that the cell count was significantly reduced by the disease, especially in patients with type 1 DM in whom the compromise was independent of the duration of the disease, and that the increase in corneal thickness in diabetic patients was greater than expected by age, regardless of the type of diabetes. These results, which clinically demonstrated the impact of hyperglycemia on the endothelium, supported the need to evaluate in vitro the effect of high glucose concentrations on these cells, particularly on their proliferation capacity, their ability to migrate to cover a defect and in the induction of apoptosis, the main type of cell death identified so far in these cells, which has been described within the responses of endothelial cells to oxidative stress, a key condition in diabetes pathophysiology. To evaluate changes in the proliferation capacity of endothelial cells, cell cultures from an immortalized line were exposed to a medium defined for them as basal and to a medium with a high concentration of glucose (55mM). The rate of change in cell density was monitored by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) reduction assays. In these experiments, cells with active metabolisms convert MTT into a purple-colored product which, once solubilized, enables any change in the number of viable cells to be evaluated by colorimetry. The tests performed showed an increase in the number of cells in the cultures exposed to high concentrations of glucose (55mM), with a significant difference after 24 hours; however, although the difference remained after 48 hours, after that time the cell count measured indirectly by the colorimetric method progressively reduced until, after 5 days, it equaled the number of viable cells of the other group. The influence of different glucose concentrations on the capacity of the endothelium to close a scratch in the monolayer was performed using the method described by Liang et al. in 2007 (Liang et al., 2007), in which a scratch is created in a cell monolayer, and photographs are then taken at regular intervals from the moment the lesion is generated until the "wound" closes. This makes it possible to compare the time it takes for the endothelial cells to close the wound under different conditions and to quantify the rate of cell migration. In these experiments, there was a significant delay in closing the defect in cells exposed to high concentrations of glucose compared to cells in basal media. While the latter took approximately 5-6 days to close the scratch, by that time cells exposed to high glucose levels had closed, on average, 50% of the distance. Finally, the evaluation of apoptosis was performed using a commercial kit (Roche Cell Death Detection ELISA PLUS) that uses the sandwich ELISA (enzyme immunoassay) technique with monoclonal antibodies for histones to determine mono- and oligonucleosomes in the cytoplasmic fraction of the cell lysates in each medium. These experiments demonstrated that exposure to high concentrations of glucose for 24 hours induced three times more apoptosis than that occurring in cells under basal conditions. In the pathophysiology of the deleterious processes associated with diabetes, the importance of electrophysiological processes has recently been identified. In renal and microglial cells, it has been shown that calcium-activated potassium channels of intermediate conductance (KCa3.1) seem to play an important role. However, the calcium-activated potassium channel (KCa) family had not previously been described in corneal endothelium and it was necessary to identify which channels of this family were expressed in these cells. We started with a bioinformatic analysis that allowed the in silico identification of these channels, after which the presence of the small conductance calcium-activated potassium channels KCa2.2 and KCa2.3, the intermediate conductance one KCa3.1 and the sodium-activated potassium channel KNa2.1 (Slick) in corneal endothelial cells was verified in vitro by PCR and in some cases by Western blot and immunolabeling. While the present work was being produced, Anumanthan et al. (2018) published a paper evaluating KCa3.1 activity in the corneal stroma that included microphotographs showing KCa3.1 expression in the endothelium, which backed up the results obtained. We proceeded to study what functions KCa3.1 has in the endothelium under basal conditions and whether these were modified when the cells were exposed to high concentrations of glucose. These experiments, which included chemical stimulation and inhibition of the channel, allowed us to identify the involvement of KCa3.1 channels in the processes of migration, proliferation and apoptosis. KCa3.1, the intermediate conductance channel of the calcium-activated potassium channel family, can be activated by 1-ethylbenzimidazolin-2-one (EBIO-1) and can be selectively inhibited by 1-[(2-Chlorophenyl)diphenylmethyl]-1H-pyrazole (TRAM-34). These compounds were used to test the effect of channel stimulation and inhibition on the proliferation, migration and apoptosis of corneal endothelial cells exposed to both basal and hyperglycemic conditions. Channel stimulation with EBIO-1 had a significant inhibitory effect on endothelial cell proliferation when used at concentrations of 50, 100 and 200 µM, sufficient to counteract the proliferative effect identified under high glucose conditions during the first few days. Inhibition of the channel with TRAM-34 at concentrations of 2, 4 and 8 µM had the opposite effect, increasing cell proliferation under basal conditions, especially at the higher concentrations, and enhancing the proliferative effect identified under high glucose conditions, maintaining a higher number of viable cells for a longer time. As for migration, stimulation with EBIO-1 reduced the migration rate by approximately 50% under basal conditions and potentiated the effect seen under hyperglycemic conditions. In contrast, inhibition of KCa3.1 with TRAM-34 at 2 µM accelerated migration and even brought the migration rate of cells under high glucose conditions closer to those of basal cells, with smaller effects at higher concentrations. Finally, under basal conditions none of the concentrations described for EBIO-1 and for TRAM-34 increased the rate of apoptosis; however, this does happen in media with high glucose concentrations, EBIO-1 at concentrations of 100 µM and TRAM_34 at 4 µM. In conclusion, this work allowed us to identify the involvement of diabetes mellitus in the corneal endothelium by determining its role in the reduction of the density of this monolayer more than that physiologically expected by age, and its impact on the increase in pachymetry, in addition to identifying a more severe involvement in patients with type 1 diabetes mellitus compared to those with type 2. Additionally, we described for the first time the presence of small conductance calcium-activated potassium channels, the expression of the intermediate conductance channel and the existence of type 2 high conductance sodium-activated potassium channels in the corneal endothelium. Finally, we explored the participation of KCa 3.1 in the proliferation, migration and apoptosis of these cells and described their role as modulators of these processes in both basal and high glucose conditions, which is relevant in both physiological and pathological conditions, not only in relation to diabetes but probably in responses to other events.es
dc.description.embargo2021-10-09 11:40:01: Script de automatizacion de embargos. Correo recibido 21 sep 2021: El día de ayer cargué en el repositorio institucional la tesis de doctorado titulada "CARACTERIZACIÓN FUNCIONAL DEL CANAL DE POTASIO ACTIVADO POR CALCIO DE CONDUCTANCIA INTERMEDIA (KCa3.1) EN EL ENDOTELIO DE LA CORNEA EN CONDICIONES FISIOLÓGICAS Y EN AMBIENTES HIPERGLÚCIDOS". He solicitado en el sistema el embargo de la misma dado que aún hay resultados pendientes de publicar y que quisiera presentarlos en artículos para revistas especializadas. Como el límite máximo de embargo son 2 años, quisiera solicitar que se mantuviera por este periodo de tiempo. Este correo tiene la intención de avisar de la marcación y las razones para ella de acuerdo con los lineamientos institucionales y preguntar si con este fin es necesario hacer alguna gestión adicional en este momento o con cierta periodicidad. Respuesta a correo 9 oct 2021: Hemos realizado la publicación de su documento: Caracterización funcional del canal de potasio activado por calcio de conductancia intermedia (KCa3.1) en el endotelio de la córnea en condiciones fisiológicas y en ambientes hiperglúcidos, el cual puede consultar en el siguiente enlace: https://repository.urosario.edu.co/handle/10336/32728 De acuerdo con su solicitud, el documento ha quedado embargado por 2 años hasta el 9 octubre de 2023 en concordancia con las Políticas de Acceso Abierto de la Universidad. Si usted desea dejarlo con acceso abierto antes de finalizar dicho periodo o si por el contrario desea extender el embargo al finalizar este tiempo, puede enviar un correo a esta misma dirección realizando la solicitud. Tenga en cuenta que los documentos en acceso abierto propician una mayor visibilidad de su producción académica y científica. De otra parte, dado que desea publicar su obra en una revista de prestigio, queremos invitarla a tomar una asesoría con nuestros asesores de información del CRAI, quienes podrán brindarle orientación en la identificación de una revista adecuada para su obra y acompañamiento en la edición para publicación. La solicitud de asesoría puede agendarla en el siguiente link: https://n9.cl/agendamiento_servicios_crai
dc.description.sponsorshipUniversidad del Rosarioes
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dc.identifier.doihttps://doi.org/10.48713/10336_32728
dc.identifier.urihttps://repository.urosario.edu.co/handle/10336/32728
dc.language.isospaes
dc.publisherUniversidad del Rosario
dc.publisher.departmentEscuela de Medicina y Ciencias de la Salud
dc.publisher.programDoctorado en Ciencias Biomédicas y Biológicas
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dc.source.instnameinstname:Universidad del Rosario
dc.source.reponamereponame:Repositorio Institucional EdocUR
dc.subjectEndotelio de la corneaes
dc.subjectDiabetes Mellituses
dc.subjectCanales de potasio activados por calcioes
dc.subjectCanales iónicoses
dc.subjectCanales de potasioes
dc.subject.ddcTecnología (Ciencias aplicadas)es
dc.subject.keywordHuman corneal endothelial celles
dc.subject.keywordPotassium channel Calcium-activatedes
dc.subject.keywordIntermediate-Conductance Calcium-Activated Potassium Channelses
dc.subject.keywordEndothelium, Corneales
dc.subject.keywordDiabetes Mellituses
dc.subject.keywordIon channelses
dc.titleCaracterización funcional del canal de potasio activado por calcio de conductancia intermedia (KCa3.1) en el endotelio de la córnea en condiciones fisiológicas y en ambientes hiperglúcidoses
dc.title.TranslatedTitleFunctional characterization of the intermediate conductance calcium-activated potassium channel (KCa3.1) in the cornea endothelium under physiological conditions and in hyperglucidic conditionses
dc.typedoctoralThesiseng
dc.type.documentTesises
dc.type.hasVersioninfo:eu-repo/semantics/acceptedVersion
dc.type.spaTesis de doctoradospa
local.department.reportEscuela de Medicina y Ciencias de la Saludspa
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CARACTERIZACION-FUNCIONAL-DEL-CANAL-DE-POTASIO-ACTIVADO-POR-CALCIO-DE-CONDUCTANCIA-INTERMEDIA-(KCa3.1)-EN-EL-ENDOTELIO-DE-LA-CORNEA-EN-CONDICIONES-FISIOLOGICAS-Y-EN-AMBIENTES-HIPERGLUCIDOS-2021.pdf
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