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: 8542166
• 负责人: JASON P WEICK
• 金额: $ 4.11万
• 依托单位: UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
• 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 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8542166
• 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.

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。完全整合需要神经元既能发送又能接收突触信息，还能根据突触连接神经元的细胞行为变化来修改它们的突触强度。由于有限的细胞跟踪和刺激技术，以前的报道只显示移植的神经元可以通过突触刺激从宿主神经元接收信息。因此，没有直接的证据表明它们能够将信息发送到宿主细胞或经历突触可塑性。为了验证这些假设，我们建议使用与人胚胎干细胞（hESC）衍生神经元中的mCherry荧光团连接的光激活视紫红质-2（ChR 2）离子通道。在将hESC衍生的前脑图案化神经元移植到小鼠海马后，我们将使用光刺激来选择性地激活人类神经元，同时记录小鼠细胞。我们假设光刺激（和随后的动作电位产生）将引起宿主神经元的强大突触激活。应用各种光刺激方案将测试人类神经元是否可以触发宿主细胞中短期和长期形式的突触可塑性。同时，我们将对ChR 2表达和非表达hESC衍生神经元的混合培养物进行类似的实验。在这里，应用光刺激方案将测试hESC衍生的神经元是否可以经历突触功效的持久变化所需的突触后变化。总之，这些数据试图提供人类神经元在宿主组织中的神经网络内充当全功能单元的能力的证据。