Institute of Neurobiology

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https://researchdata.hhu.de/handle/123456789/133
The Institute of Neurobiology at the HHU is devoted to the elucidation and study of intracellular ion dynamics in the vertebrate brain under physiological as well as pathophysiological conditions.

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    Petersilie et al. 2024b - Figure 1
    (STAR Protocols, 2024) Petersilie, Laura; Kafitz, Karl W.; Neu, Louis A.; Heiduschka, Sonja; Le, Stephanie; Prigione, Alessandro; Rose, Christine R.
    Three-dimensional brain organoids from human pluripotent stem cells are a powerful tool for studying human neural networks. This article presents a refined protocol for generating robust brain organoid slices derived from regionalized cortical organoids and grown at the air-liquid interphase. The procedures for slicing organoids and maintaining them in long-term culture are described. We then detail approaches for quality control including evaluation of cell death and cellular identity. Finally, we describe procedures for expression of a genetically-encoded nanosensor for ATP.
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    Petersilie et al. 2024b - Figure 2
    (STAR Protocols, 2024) Petersilie, Laura; Kafitz, Karl W.; Neu, Louis A.; Heiduschka, Sonja; Le, Stephanie; Prigione, Alessandro; Rose, Christine R.
    Three-dimensional brain organoids from human pluripotent stem cells are a powerful tool for studying human neural networks. This article presents a refined protocol for generating robust brain organoid slices derived from regionalized cortical organoids and grown at the air-liquid interphase. The procedures for slicing organoids and maintaining them in long-term culture are described. We then detail approaches for quality control including evaluation of cell death and cellular identity. Finally, we describe procedures for expression of a genetically-encoded nanosensor for ATP.
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    Petersilie et al. 2024 - Supplementary Figure
    (iScience, 2024) Petersilie, Laura; Heiduschka, Sonja; Nelson, Joel S. E.; Neu, Louis A.; Le, Stephanie; Anand, Ruchika; Kafitz, Karl W.; Prigione, Alessandro; Rose, Christine R.
    Brain organoids derived from human pluripotent stem cells are a promising tool for studying human neurodevelopment and related disorders. Here, we generated long-term cultures of cortical brain organoid slices (cBOS) grown at the air-liquid interphase from regionalized cortical organoids. We show that cBOS host mature neurons and astrocytes organized in complex architecture. Whole-cell patch clamp demonstrated subthreshold synaptic inputs and action potential firing of neurons. Spontaneous intracellular calcium signals turned into synchronous large-scale oscillations upon combined disinhibition of NMDA receptors and blocking of GABAA receptors. Brief metabolic inhibition to mimic transient energy restriction in the ischemic brain induced reversible intracellular calcium loading of cBOS. Moreover, metabolic inhibition induced a reversible decline in neuronal ATP as revealed by ATeam1.03YEMK. Overall, cBOS provide a powerful platform to assess morphological and functional aspects of human neural cells in intact minimal networks and to address the pathways that drive cellular damage during brain ischemia.
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    Petersilie et al. 2024 - Figure 7
    (iScience, 2024) Petersilie, Laura; Heiduschka, Sonja; Nelson, Joel S. E.; Neu, Louis A.; Le, Stephanie; Anand, Ruchika; Kafitz, Karl W.; Prigione, Alessandro; Rose, Christine R.
    Brain organoids derived from human pluripotent stem cells are a promising tool for studying human neurodevelopment and related disorders. Here, we generated long-term cultures of cortical brain organoid slices (cBOS) grown at the air-liquid interphase from regionalized cortical organoids. We show that cBOS host mature neurons and astrocytes organized in complex architecture. Whole-cell patch clamp demonstrated subthreshold synaptic inputs and action potential firing of neurons. Spontaneous intracellular calcium signals turned into synchronous large-scale oscillations upon combined disinhibition of NMDA receptors and blocking of GABAA receptors. Brief metabolic inhibition to mimic transient energy restriction in the ischemic brain induced reversible intracellular calcium loading of cBOS. Moreover, metabolic inhibition induced a reversible decline in neuronal ATP as revealed by ATeam1.03YEMK. Overall, cBOS provide a powerful platform to assess morphological and functional aspects of human neural cells in intact minimal networks and to address the pathways that drive cellular damage during brain ischemia.
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    Petersilie et al. 2024 - Figure 6
    (iScience, 2024) Petersilie, Laura; Heiduschka, Sonja; Nelson, Joel S. E.; Neu, Louis A.; Le, Stephanie; Anand, Ruchika; Kafitz, Karl W.; Prigione, Alessandro; Rose, Christine R.
    Brain organoids derived from human pluripotent stem cells are a promising tool for studying human neurodevelopment and related disorders. Here, we generated long-term cultures of cortical brain organoid slices (cBOS) grown at the air-liquid interphase from regionalized cortical organoids. We show that cBOS host mature neurons and astrocytes organized in complex architecture. Whole-cell patch clamp demonstrated subthreshold synaptic inputs and action potential firing of neurons. Spontaneous intracellular calcium signals turned into synchronous large-scale oscillations upon combined disinhibition of NMDA receptors and blocking of GABAA receptors. Brief metabolic inhibition to mimic transient energy restriction in the ischemic brain induced reversible intracellular calcium loading of cBOS. Moreover, metabolic inhibition induced a reversible decline in neuronal ATP as revealed by ATeam1.03YEMK. Overall, cBOS provide a powerful platform to assess morphological and functional aspects of human neural cells in intact minimal networks and to address the pathways that drive cellular damage during brain ischemia.
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    Petersilie et al. 2024b - Figure 3
    (STAR Protocols, 2024) Petersilie, Laura; Kafitz, Karl W.; Neu, Louis A.; Heiduschka, Sonja; Le, Stephanie; Prigione, Alessandro; Rose R., Christine
    Three-dimensional brain organoids from human pluripotent stem cells are a powerful tool for studying human neural networks. This article presents a refined protocol for generating robust brain organoid slices derived from regionalized cortical organoids and grown at the air-liquid interphase. The procedures for slicing organoids and maintaining them in long-term culture are described. We then detail approaches for quality control including evaluation of cell death and cellular identity. Finally, we describe procedures for expression of a genetically-encoded nanosensor for ATP.
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    Petersilie et al. 2024 - Figure 3
    (iScience, 2024) Petersilie, Laura; Heiduschka, Sonja; Nelson, Joel S. E.; Neu, Louis A.; Le, Stephanie; Anand, Ruchika; Kafitz, Karl W.; Prigione, Alessandro; Rose, Christine R.
    Brain organoids derived from human pluripotent stem cells are a promising tool for studying human neurodevelopment and related disorders. Here, we generated long-term cultures of cortical brain organoid slices (cBOS) grown at the air-liquid interphase from regionalized cortical organoids. We show that cBOS host mature neurons and astrocytes organized in complex architecture. Whole-cell patch clamp demonstrated subthreshold synaptic inputs and action potential firing of neurons. Spontaneous intracellular calcium signals turned into synchronous large-scale oscillations upon combined disinhibition of NMDA receptors and blocking of GABAA receptors. Brief metabolic inhibition to mimic transient energy restriction in the ischemic brain induced reversible intracellular calcium loading of cBOS. Moreover, metabolic inhibition induced a reversible decline in neuronal ATP as revealed by ATeam1.03YEMK. Overall, cBOS provide a powerful platform to assess morphological and functional aspects of human neural cells in intact minimal networks and to address the pathways that drive cellular damage during brain ischemia.
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    Petersilie et al. 2024 - Figure 4
    (iScience, 2024) Petersilie, Laura; Heiduschka, Sonja; Nelson, Joel S. E.; Neu, Louis A.; Le, Stephanie; Anand, Ruchika; Kafitz, Karl W.; Prigione, Alessandro; Rose, Christine R.
    Brain organoids derived from human pluripotent stem cells are a promising tool for studying human neurodevelopment and related disorders. Here, we generated long-term cultures of cortical brain organoid slices (cBOS) grown at the air-liquid interphase from regionalized cortical organoids. We show that cBOS host mature neurons and astrocytes organized in complex architecture. Whole-cell patch clamp demonstrated subthreshold synaptic inputs and action potential firing of neurons. Spontaneous intracellular calcium signals turned into synchronous large-scale oscillations upon combined disinhibition of NMDA receptors and blocking of GABAA receptors. Brief metabolic inhibition to mimic transient energy restriction in the ischemic brain induced reversible intracellular calcium loading of cBOS. Moreover, metabolic inhibition induced a reversible decline in neuronal ATP as revealed by ATeam1.03YEMK. Overall, cBOS provide a powerful platform to assess morphological and functional aspects of human neural cells in intact minimal networks and to address the pathways that drive cellular damage during brain ischemia.
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    Petersilie et al. 2024 - Figure 5
    (iScience, 2024) Petersilie, Laura; Heiduschka, Sonja; Nelson, Joel S. E.; Neu, Louis A.; Le, Stephanie; Anand, Ruchika; Kafitz, Karl W.; Prigione, Alessandro; Rose, Christine R.
    Brain organoids derived from human pluripotent stem cells are a promising tool for studying human neurodevelopment and related disorders. Here, we generated long-term cultures of cortical brain organoid slices (cBOS) grown at the air-liquid interphase from regionalized cortical organoids. We show that cBOS host mature neurons and astrocytes organized in complex architecture. Whole-cell patch clamp demonstrated subthreshold synaptic inputs and action potential firing of neurons. Spontaneous intracellular calcium signals turned into synchronous large-scale oscillations upon combined disinhibition of NMDA receptors and blocking of GABAA receptors. Brief metabolic inhibition to mimic transient energy restriction in the ischemic brain induced reversible intracellular calcium loading of cBOS. Moreover, metabolic inhibition induced a reversible decline in neuronal ATP as revealed by ATeam1.03YEMK. Overall, cBOS provide a powerful platform to assess morphological and functional aspects of human neural cells in intact minimal networks and to address the pathways that drive cellular damage during brain ischemia.
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    Petersilie et al. 2024b - Figure 2
    (STAR Protocols, 2024) Petersilie, Laura; Kafitz, Karl W.; Neu, Louis A.; Heiduschka, Sonja; Le, Stephanie; Prigione, Alessandro; Rose, Christine R.
    Three-dimensional brain organoids from human pluripotent stem cells are a powerful tool for studying human neural networks. This article presents a refined protocol for generating robust brain organoid slices derived from regionalized cortical organoids and grown at the air-liquid interphase. The procedures for slicing organoids and maintaining them in long-term culture are described. We then detail approaches for quality control including evaluation of cell death and cellular identity. Finally, we describe procedures for expression of a genetically-encoded nanosensor for ATP.
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    Petersilie et al. 2024 - Figure 2
    (iScience, 2024) Petersilie, Laura; Heiduschka, Sonja; Nelson, Joel S. E.; Neu, Louis A.; Le, Stephanie; Anand, Ruchika; Kafitz, Karl W.; Prigione, Alessandro; Rose, Christine R.
    Brain organoids derived from human pluripotent stem cells are a promising tool for studying human neurodevelopment and related disorders. Here, we generated long-term cultures of cortical brain organoid slices (cBOS) grown at the air-liquid interphase from regionalized cortical organoids. We show that cBOS host mature neurons and astrocytes organized in complex architecture. Whole-cell patch clamp demonstrated subthreshold synaptic inputs and action potential firing of neurons. Spontaneous intracellular calcium signals turned into synchronous large-scale oscillations upon combined disinhibition of NMDA receptors and blocking of GABAA receptors. Brief metabolic inhibition to mimic transient energy restriction in the ischemic brain induced reversible intracellular calcium loading of cBOS. Moreover, metabolic inhibition induced a reversible decline in neuronal ATP as revealed by ATeam1.03YEMK. Overall, cBOS provide a powerful platform to assess morphological and functional aspects of human neural cells in intact minimal networks and to address the pathways that drive cellular damage during brain ischemia.
  • No Thumbnail Available
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    Petersilie et al. 2024 - Figure 1
    (iScience, 2024) Petersilie, Laura; Heiduschka, Sonja; Nelson, Joel S. E.; Neu, Louis A.; Le, Stephanie; Anand, Ruchika; Kafitz, Karl W.; Prigione, Alessandro; Rose, Christine R.
    Brain organoids derived from human pluripotent stem cells are a promising tool for studying human neurodevelopment and related disorders. Here, we generated long-term cultures of cortical brain organoid slices (cBOS) grown at the air-liquid interphase from regionalized cortical organoids. We show that cBOS host mature neurons and astrocytes organized in complex architecture. Whole-cell patch clamp demonstrated subthreshold synaptic inputs and action potential firing of neurons. Spontaneous intracellular calcium signals turned into synchronous large-scale oscillations upon combined disinhibition of NMDA receptors and blocking of GABAA receptors. Brief metabolic inhibition to mimic transient energy restriction in the ischemic brain induced reversible intracellular calcium loading of cBOS. Moreover, metabolic inhibition induced a reversible decline in neuronal ATP as revealed by ATeam1.03YEMK. Overall, cBOS provide a powerful platform to assess morphological and functional aspects of human neural cells in intact minimal networks and to address the pathways that drive cellular damage during brain ischemia.
  • No Thumbnail Available
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    Petersilie et al. 2024b - Figure 5
    (STAR Protocols, 2024) Petersilie, Laura; Kafitz, Karl W.; Neu, Louis A.; Heiduschka, Sonja; Le, Stephanie; Prigione, Alessandro; Rose, Christine R.
    Three-dimensional brain organoids from human pluripotent stem cells are a powerful tool for studying human neural networks. This article presents a refined protocol for generating robust brain organoid slices derived from regionalized cortical organoids and grown at the air-liquid interphase. The procedures for slicing organoids and maintaining them in long-term culture are described. We then detail approaches for quality control including evaluation of cell death and cellular identity. Finally, we describe procedures for expression of a genetically-encoded nanosensor for ATP.
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    Petersilie et al. 2024b - Figure 4
    (STAR Protocols, 2024) Petersilie, Laura; Kafitz, Karl W.; Neu, Louis A.; Heiduschka, Sonja; Le, Stephanie; Prigione, Alessandro; Rose, Christine R.
    Three-dimensional brain organoids from human pluripotent stem cells are a powerful tool for studying human neural networks. This article presents a refined protocol for generating robust brain organoid slices derived from regionalized cortical organoids and grown at the air-liquid interphase. The procedures for slicing organoids and maintaining them in long-term culture are described. We then detail approaches for quality control including evaluation of cell death and cellular identity. Finally, we describe procedures for expression of a genetically-encoded nanosensor for ATP.