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，我们研究了视黄酸在神经元再生过程中以及突触变化（可塑性）过程中对神经突生长（和生长锥转动行为）的新的非基因组效应。为此，我们利用已鉴定的软体动物 Lymnaea stagnalis 的成年神经元，因为这些神经元在体外再生并重建特定的突触连接。 我的研究项目成功的关键是一套记录套件，其中包括配备荧光功能的蔡司倒置显微镜、精密显微操作器和电记录设备，可对培养的神经元、神经突或突触连接的细胞对进行同步电生理测量。该设备现已使用 20 多年（有些更旧），要么不再正常运行，要么完全失效。 该录音设备是布鲁克大学唯一的此类录音设备。除了我自己的研究生和本科生 (HQP) 每天要求使用之外，我在布罗克 (Brock) 的合作者（及其 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项目的进展、我的总部的进度和产出，并对我的合作产生了负面影响。因此，我要求升级该显微镜的成像功能，以便提供更合适和更复杂的功能来对培养神经元中的各种信号分子（microRNA、mRNAS 和标记蛋白）进行成像。我还迫切需要新的精密显微操作器和电记录设备来继续我们关于视黄酸对离子通道活动和突触功能影响的电生理学研究。 过去 20 年在我的实验室接受过培训的所有研究生，以及许多荣誉论文本科生和 NSERC-USRA 学生，都接受过培训并使用过该记录和成像设备。作为我所在系（实际上是数学与科学学院）唯一的女性神经生物学家，我是女学生的导师，并在我的研究项目的各个方面大力促进公平、多样性和包容性。 总之，如果没有所需的设备，该装备将无法再按预期运行，并将危及我进行最近的 NSERC 发现拨款（2021-2026）中概述的实验的能力以及我继续训练 HQP 的能力。我的实验室进行的所有研究均由 NSERC 资助，并专门为 NSE 做出贡献。