Functional integration and plasticity of human ESC-derived neurons

人类ESC衍生神经元的功能整合和可塑性

• 批准号: 8097001
• 负责人: JASON P WEICK
• 金额: $ 7.43万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-01 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8097001
• 关键词: Action Potentials; Acute; Animals; Behavior; Biological Neural Networks; Brain; Cations; Cell Transplants; Cells; Coculture Techniques; Data; Fluorescence; Frequencies; Generations; Health; Hippocampus (Brain); Human; Immune response; Ion Channel; Knowledge; Left; Light; Link; Long-Term Depression; Long-Term Potentiation; Mental Depression; Methods; Mus; Neuraxis; Neurons; Output; Pattern; Physiologic pulse; Physiological; Property; Prosencephalon; Protocols documentation; Replacement Therapy; Reporting; Research; Rodent; Slice; Specificity; Stem cells; Stimulus; Synapses; Synaptic plasticity; Techniques; Testing; Therapeutic Effect; Tissues; Transplantation; blastomere structure; extracellular; fluorophore; human embryonic stem cell; human embryonic stem cell transplantation; millisecond; neuronal excitability; neuronal replacement; novel; postsynaptic; presynaptic; prevent; research study; response; success; tool

DESCRIPTION (provided by applicant): The success of neuronal cell replacement therapy depends on the ability of transplanted cells to synaptically integrate with host tissue. Complete integration requires that neurons can both send and receive synaptic information, as well as to modify their synaptic strength in response to changes in the cellular behavior of synaptically connected neurons. Due to limited cell tracking and stimulation techniques, previous reports have shown only that transplanted neurons can receive information from host neurons via synaptic stimulation. Thus, no direct evidence exists for their ability to send information to host cells or undergo synaptic plasticity. To test these hypotheses, we propose to use the light-activated Channelrhodopsin-2 (ChR2) ion channel linked to the mCherry fluorophore in human embryonic stem cell (hESC)-derived neurons. Following transplantation of hESC-derived forebrain-patterned neurons to the mouse hippocampus, we will use light stimulation to selectively activate human neurons while recording mouse cells. We hypothesize that light stimulation (and subsequent action potential generation) will give rise to robust synaptic activation of host neurons. Application of various light stimulus protocols will then test whether human neurons can trigger short-term and long-term forms of synaptic plasticity in host cells. In parallel, we will perform similar experiments on mixed cultures of ChR2-expressing and non-expressing hESC-derived neurons. Here, application of light stimulation protocols will test whether hESC-derived neurons can undergo post-synaptic changes required for enduring changes in synaptic efficacy. Together, these data seek to provide evidence of the ability of human neurons to act as a fully functional unit within a neural network in host tissues. PUBLIC HEALTH RELEVANCE: Successful neuronal cell replacement is thought to rely on the integration of transplanted cells with host tissue via the formation of synaptic connections. Until recently, technical limitations have prevented the determination of whether stem cell-derived neurons can send information to host cells or undergo changes in synaptic efficacy that are necessary for true circuit integration. The proposed research will use the newly characterized light activated Channelrhodopsin-2 in human embryonic cell-derived neurons to regulate the excitability of transplanted neurons to test these hypotheses.

描述（由申请人提供）：神经元细胞替代疗法的成功取决于移植细胞与宿主组织突触整合的能力。完整的整合要求神经元可以发送和接收突触信息，并可以改变其突触强度，以应对突触连接神经元的细胞行为的变化。由于细胞跟踪和刺激技术有限，先前的报道仅表明，移植神经元可以通过突触刺激从宿主神经元接收信息。因此，没有直接证据证明其向宿主细胞发送信息或经历突触可塑性的能力。为了检验这些假设，我们建议使用与人类胚胎干细胞（HESC）衍生的神经元中麦克里荧光团相关的光激活通道Ropopopsin-2（ChR2）离子通道。将hESC衍生的前脑形式神经元移植到小鼠海马后，我们将使用光刺激在记录小鼠细胞时选择性激活人神经元。我们假设光刺激（以及随后的动作电位产生）会导致宿主神经元的强大突触激活。然后，应用各种光刺激方案的应用将测试人类神经元是否可以触发宿主细胞中突触可塑性的短期和长期形式。同时，我们将对表达CHR2和非表达hESC衍生的神经元的混合培养物进行类似的实验。在这里，光刺激方案的应用将测试hESC衍生的神经元是否可以经历持久突触功效变化所需的突触后变化。这些数据共同寻求提供证据表明人神经元在宿主组织中神经网络中充当功能齐全的单位的能力。 公共卫生相关性：成功的神经元细胞置换被认为依赖于通过形成突触连接的移植细胞与宿主组织的整合。直到最近，技术局限性仍阻止了干细胞衍生的神经元是否可以向宿主细胞发送信息或经历真正电路积分所需的突触功效的变化。拟议的研究将在人类胚胎细胞衍生的神经元中使用新表征的光活化通道Ropopostin-2来调节移植神经元的兴奋性以检验这些假设。