ABSTRACT
Genetic abnormalities affecting glutamate receptors are central to excitatory overload-driven neuronal mechanisms that culminate in seizures, making them pivotal targets in epilepsy research. Increasingly used to advance this field, the genetically audiogenic seizure hamster from Salamanca (GASH/Sal) exhibits generalized seizures triggered by high-intensity acoustic stimulation and harbors significant genetic variants recently identified through whole-exome sequencing. Here, we addressed the influence of the missense single-nucleotide polymorphism (C9586732T, p.His289Tyr) in the glutamate receptor ionotropic kainate-1 (Grik1) gene and its implications for the GASH/Sal seizure susceptibility. Using a protein 3D structure prediction, we showed a potential effect of this sequence variation, located in the amino-terminal domain, on the stability and/or conformation of the kainate receptor subunit-1 protein (GluK1). We further employed a multi-technique approach, encompassing gene expression analysis (RT-qPCR), Western blotting, and immunohistochemistry in bright-field and confocal fluorescence microscopy, to investigate critical seizure-associated brain regions in GASH/Sal animals under seizure-free conditions compared to matched wild-type controls. We detected disruptions in the transcriptional profile of the Grik1 gene within the audiogenic seizure-associated neuronal network. Alterations in GluK1 protein levels were also observed in various brain structures, accompanied by an unexpected lower molecular weight band in the inferior and superior colliculi. This correlated with substantial disparities in GluK1-immunolabeling distribution across multiple brain regions, including the cerebellum, hippocampus, subdivisions of the inferior and superior colliculi, and the prefrontal cortex. Notably, the diffuse immunolabeling accumulated within perikarya, axonal fibers and terminals, exhibiting a prominent concentration in proximity to the cell nucleus. This suggests potential disturbances in the GluK1-trafficking mechanism, which could subsequently affect glutamate synaptic transmission. Overall, our study sheds light on the genetic underpinnings of seizures and underscores the importance of investigating the molecular mechanisms behind synaptic dysfunction in epileptic neural networks, laying a crucial foundation for future research and therapeutic strategies targeting GluK1-containing kainate receptors.
ABSTRACT
La absorción del Real Tribunal del Protoalbeitarato1 por la Escuela de Veterinaria de Madrid presenta ciertas singularidades respecto al proceso equivalente en medicina y farmacia. La pugna y hegemonía del Real Tribunal durante casi sesenta años desde la creación de la Escuela de Veterinaria (1792), ha sido estudiada por la historiografía tradicional con una visión centrada fundamentalmente en aspectos veterinarios. En el presente artículo se investigan en detalle los procesos conducentes a la absorción y se pone de manifiesto la relevancia de las motivaciones políticas y su predominio sobre las de carácter técnico, sanitario o profesional. Por ello, se analiza el juego de instituciones y de agentes externos a la veterinaria implicados, en especial destacamos el papel desempeñado por el XV Marqués de Cerralbo (AU)
The absorption of the Real Tribunal del Protoalbeitarato by the Veterinary Science School of Madrid shows several singularities in relation to the equivalent process in medicine and pharmacy.The conflict and the hegemony of the Real Tribunal during almost sixty years from the creation of the Veterinary Science School (1792), has been studied by the traditional historiography with a view fundamentally focused on veterinary aspects. In this article, the process of absorption is investigated in full detail. Also, the relevance of political motivations and their predominance over technical, sanitary or professional motivations are shown. So, the game of institutions and agents which are external to the veterinary science are analysed; specially, the out standing role performed by the XV Marquis of Cerralbo(AU)