Optogenetic Mapping of Synaptic Connections in Mouse Brain Slices to Define the Functional Connectome of Identified Neuronal Populations
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Author:
Date:
2017-01-05
[Abstract] Functional connectivity in a neural circuit is determined by the strength, incidence, and neurotransmitter nature of its connections (Chuhma, 2015). Using optogenetics the functional synaptic connections between an identified population of neurons and defined postsynaptic target neurons may be measured systematically in order to determine the functional connectome of that identified population. Here we describe the experimental protocol used to investigate the excitatory functional connectome of ventral midbrain dopamine neurons, mediated by glutamate cotransmission (Mingote et al., 2015). Dopamine neurons are made light sensitive by injecting an adeno-associated virus (AAV) encoding channelrhodopsin (ChR2) into the ventral midbrain of DATIREScre mice. The efficacy and ...
[摘要] 神经回路中的功能连通性由其连接的强度,发病率和神经递质特性决定(Chuhma,2015)。使用光遗传学,可以系统地测量识别的神经元群体和定义的突触后靶神经元之间的功能性突触连接,以便确定该识别群体的功能性连接群。这里我们描述了用于研究由谷氨酸共转播介导的腹侧中脑多巴胺神经元的兴奋性功能性连接体的实验方案(Mingote等,2015)。通过将编码通道视紫红质(ChR2)的腺相关病毒(AAV)注射到DATIREScre小鼠的腹侧中脑中,使多巴胺神经元变得光敏。多巴胺合成酶酪氨酸羟化酶的免疫荧光证实ChR2表达在多巴胺神经元中的功效和特异性。然后,切片膜片钳记录由接受多巴胺神经元投影的区域中的神经元产生,并且确定兴奋性连接的发生率和强度。所有接受多巴胺神经元投射的区域的连接发生率和强度的总结构成功能性连接体。 【背景】为了建立特定神经回路的功能,有必要确定解剖连接,解剖连接的映射及其功能连接,连接的强度,发病率和神经递质性质的映射。使用单因素限制的病毒性突触后追踪技术,可以描述包括多巴胺系统在内的神经回路的复杂解剖连接(Callaway and Luo,2015; ...
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Two-event Transfusion-related Acute Lung Injury Mouse Model
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Author:
Date:
2015-06-20
[Abstract] Transfusion-related acute lung injury (TRALI) is defined as acute lung injury that occurs within 6 hours of a blood product transfusion. TRALI continues to be a leading cause of transfusion-related mortality and we have developed a mouse model of TRALI to better understand the mechanisms by which injury occurs and to test therapeutic approaches. Our model is a two-event model based on immune priming and the challenge of BALB/c wild-type mice with cognate MHC Class I monoclonal antibody (MHC I mAb). Immune priming with LPS mimics the primed state of recipients (first event) that is important for the development of TRALI. Donor HLA antibodies are frequently implicated in TRALI reactions, and cognate MHC Class I antibody (second event) produces acute lung injury in primed animals. Here, we ...
[摘要] 输血相关的急性肺损伤(TRALI)定义为在血液制品输血的6小时内发生的急性肺损伤。 TRALI仍然是输血相关死亡率的主要原因,我们已经开发了TRALI的小鼠模型,以更好地了解损伤发生的机制和测试治疗方法。 我们的模型是基于免疫引发的双事件模型和具有同源MHC I类单克隆抗体(MHC I mAb)的BALB/c野生型小鼠的攻击。 用LPS的免疫引发模拟对TRALI的发展重要的受体的引发状态(第一事件)。 供体HLA抗体经常涉及TRALI反应,并且同源MHC I类抗体(第二事件)在引发的动物中产生急性肺损伤。 在这里,我们描述一个详细的协议,在动物中具有高重现性。
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