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资助的研究项目研究维甲酸(维生素A的代谢物)对神经系统功能的影响。特别是，我们研究了维甲酸在神经元再生和突触变化(可塑性)过程中对轴突生长(和生长锥体转向行为)的新的非基因组效应。为此，我们利用已鉴定的软体动物Lymneea stagnaris的成年神经元，因为这些神经元在体外再生和改造特定的突触连接。我的研究项目成功的关键是一个记录套件，它包括一个配备有荧光功能的蔡司倒置显微镜，以及精密的微操作器和电子记录设备，允许对培养的神经元、神经突起或突触连接的细胞对进行同步电生理测量。该设备现在已有20多年的历史(有些更老)，要么不再正常工作，要么完全失效。这台录音机是布罗克大学唯一一台这样的录音机。除了我自己的研究生和本科生(HQP)每天都需要访问之外，我在BROCK的合作者(和他们的HQP)也可以访问。目前，无法更换旧的和故障的设备严重阻碍了我整个研究计划的进度，阻碍了我的HQP的进度和产出，并对我的合作产生了负面影响。因此，我要求升级这台显微镜的成像能力，以便提供更合适和更复杂的能力来成像培养神经元中的各种信号分子(microRNAs、mRNAs和标记蛋白)。我还迫切需要新的精密显微操纵器和电记录设备来继续我们对维甲酸对离子通道活动和突触功能影响的电生理学研究。在过去的20年里，所有在我的实验室接受培训的研究生，以及许多荣誉论文本科生和NSERC-USRA学生，都接受过这种录音和成像设备的培训，并使用过这种设备。作为我所在系(甚至是数学与科学学院)唯一的女性神经生物学家，我是女学生的导师，并在我的研究计划的各个方面大力促进公平、多样性和包容性。总而言之，如果没有所需的设备，这个钻井平台将不再按预期运行，并将危及我执行最近的NSERC发现拨款(2021-2026年)中概述的实验的能力，以及我继续培训HQP的能力。在我的实验室进行的所有研究都是由NSERC资助的，并且只对NSE做出贡献。