Essential upgrade to a cell culture imaging and recording rig to study the role of retinoids (Vitamin A metabolites) in nervous system function

对细胞培养成像和记录装置进行必要升级，以研究类视黄醇（维生素 A 代谢物）在神经系统功能中的作用

• 批准号: RTI-2022-00225
• 负责人: Spencer, Gaynor
• 金额: $ 7.83万
• 依托单位: Brock University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=742930
• 关键词: Essential; upgrade; cell; culture; imaging

My NSERC-funded research program studies the effects of retinoic acid (the metabolite of Vitamin A) on nervous system function. In particular, we study novel non-genomic effects of retinoic acid on neurite outgrowth (and growth cone turning behaviour) during neuronal regeneration, as well as during synaptic changes (plasticity). To this end, we utilize identified adult neurons of the mollusc Lymnaea stagnalis, as these neurons regenerate and reform specific synaptic connections in vitro. Key to the success of my research program is a recording suite that consists of a Zeiss inverted microscope equipped with fluorescence capabilities, as well as precision micromanipulators and electrical recording equipment to allow for simultaneous electrophysiological measurements from cultured neurons, neurites or synaptically connected cell pairs. This equipment is now over 20 years old (some is older) and is either no longer functioning properly or has completely failed. This recording rig is the only one of its kind at Brock University. As well as being required daily by my own graduate and undergraduate students (HQP), my collaborators at Brock (and their HQP) also have access. At present, the inability to replace the old and failed equipment is severely hindering the progress of my entire research program, the progress and output of my HQP, and is negatively affecting my collaborations. I am thus requesting upgrades to the imaging capabilities of this microscope, in order to provide more suitable and sophisticated capabilities to image various signaling molecules in cultured neurons (microRNAs, mRNAS and tagged proteins). I also desperately require new precision micromanipulators and electrical recording equipment to continue our electrophysiological studies of the effects of retinoic acid on ion channel activity and synaptic function. All of the graduate students trained in my lab over the last 20 years, as well as many Honours thesis undergraduate students and NSERC-USRA students, have been trained on and have used this recording and imaging rig. As the only female Neurobiologist in my department (and indeed the Faculty of Math and Science), I am a mentor to female students and strongly promote equity, diversity and inclusion in all aspects of my research program. In summary, without the requested equipment, this rig will no longer be functioning as intended, and will jeopardize my ability to carry out the experiments outlined in my recent NSERC Discovery grant (2021-2026) as well as my ability to continue to train HQP. All of the research carried out in my lab is funded by NSERC and contributes solely to NSE.

我的NSERC资助研究计划研究了视黄酸（维生素A的代谢产物）对神经系统功能的影响。特别是，我们研究了视黄酸对神经元再生期间神经突生长（以及生长锥行为）的新型非基因组作用，以及在突触变化（可塑性）期间。为此，我们利用了软体动物淋巴结杆菌的鉴定的成年神经元，因为这些神经元在体外再生并改革了特定的突触连接。 我的研究计划成功的关键是一个录音套件，该套件由配备有荧光功能的Zeiss倒置显微镜以及精确的显微镜和电记录设备，以允许从培养的神经元，神经元，神经元，神经元或突触连接的电池组中同时进行电生理测量。该设备现在已有20多年的历史了（有些年龄较大），要么不再正常运行，要么完全失败。 该录音台是布罗克大学唯一同类的钻机。除了我自己的毕业生和本科生（HQP）每天都需要，我在Brock（及其HQP）的合作者也可以访问。目前，无法替换旧设备和失败的设备严重阻碍了我整个研究计划，HQP的进度和输出的进步，并且对我的协作产生了负面影响。因此，我要求升级该显微镜的成像功能，以便提供更合适和复杂的功能，以在培养的神经元（microRNA，mRNA和标记蛋白）中成像各种信号分子。我还迫切需要新的精密微型手持和电记录设备，以继续我们对视黄酸对离子通道活性和突触功能的影响的电生理研究。 在过去的20年中，所有在我的实验室中接受过培训的研究生，以及许多荣誉论文的本科生和NSERC-ISRA学生，都接受过培训，并使用了此录音和成像钻机。作为我系中唯一的女性神经生物学家（实际上是数学和科学学院），我是女学生的导师，并在我的研究计划的各个方面都强烈促进公平，多样性和包容性。 总而言之，如果没有请求的设备，该钻机将不再按预期运行，并且会危及我在最近的NSERC Discovery Grant（2021-2026）中概述的实验的能力，以及我继续培训HQP的能力。在我的实验室中进行的所有研究都由NSERC资助，仅为NSE做出贡献。