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Dissecting complexity: Discovering the micro-organization and microconnectivity of unguided cortical organoids
TORRES-VAZQUEZ, IRMA IRIS
TORRES-VAZQUEZ, IRMA IRIS
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Abstract
The primary goal is to provide techniques and resources for understanding and delineating the structure of complex circuits at the level of synaptic connections, alone or in combination with reagents, tools, and protocols for identifying important cellular and circuit features, for example, for classifying or characterizing cellular or synaptic phenotypes and components.
The synaptic components and interaction between nerve cells is critical for the development and establishment of functional microconnectivity between brain nerves. Both unidirectional chemical synaptic transmission and bi-directional electrical transmission are found in the brain. However, the study of synaptic transmission between nerve cells in the human nervous system has focused primarily on the unidirectional chemical synapse, with little to no focus on the bi-directional electrical synapse. In electrical synapses, gap junctions are the basic structure; they are clusters of intercellular channels that allow the exchange of small molecules and ions between two opposing cells. Connexins are the structural protein of gap junctions, while connexin 36 is the only neuronal connexin in humans. Despite the increasing numbers of studies on gap junctional–mediated-coupling, the role of gap junctions and Connexin 36 (Cx 36) in development and diseases is limited. Cortical organoids are three-dimensional, in vitro culture models that offer a unique platform to study the complexity of cellular and molecular interactions between nerve cells. We developed unguided human cortical organoids derived from pluripotent stem cells to characterize the unidirectional chemical and bi-directional electrical synapses at 30-, 60-, and 90-day old cortical organoids using whole-mount fluorescence immunohistochemistry. We characterized cellular components that could describe the micro-organization of these cortical organoids. In addition, our functional microconnectivity analysis showed the presence of both unidirectional chemical synapses and bi-directional electrical synapses. In the specific case of electrical synapses, Cx 36 and a phosphorylated form of Cx 36 (were found in these human unguided cortical organoids at all time points.
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2025-01-01
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University of Kansas
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TORRES-VAZQUEZ_ku_0099D_19714.pdf
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Neurosciences