Research Initiation Award: Defining the role of DJ-1 in regulating L-type voltage-dependent calcium channel expression in neuronal plasticity

研究启动奖：定义 DJ-1 在调节神经元可塑性中 L 型电压依赖性钙通道表达中的作用

• 批准号: 2200474
• 负责人: Farr Niere
• 金额: $ 30万
• 依托单位: North Carolina Agricultural & Technical State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2200474&HistoricalAwards=false
• 关键词: Research; Initiation; Award; Defining; role

The Historically Black Colleges and Universities Undergraduate Program (HBCU-UP) through Research Initiation Awards provide support for junior and mid-career faculty at Historically Black Colleges and Universities who are building new research programs or redirecting and rebuilding existing research programs. It is expected that the award helps to further the faculty member's research capability and effectiveness and improve research and teaching at the home institution. This award to North Carolina A & T State University provides an opportunity to establish a molecular neuroscience laboratory that investigates the mechanisms underlying learning and memory. Specifically, the project aims to examine how L-type calcium ion channels, proteins that allow calcium to enter a neuron, are prevented from being made during long-term synaptic depression, which is a form of neuronal plasticity. With this award, efforts also include the training of undergraduate students thereby contributing to the next generation of scientists who can contribute to scientific innovation, creativity and productivity.Encoding learning and memory, such as the ability to distinguish between new and old experiences, requires neurons to synthesize some and repress other proteins in synapses and dendrites. One mechanism underlying the ability of neurons to regulate their protein levels rapidly is through RNA-binding proteins (RBPs). However, current knowledge about the identity of these RBPs, their messenger RNA (mRNA) targets, and how these RBPs regulate the protein expression of their targets are poorly understood. Recent studies revealed that DJ-1, an under-characterized RBP, resides in dendrites and synapses, and modulates the activity of some dendritic calcium channels. Thus, this project seeks to investigate the role of DJ-1 in regulating the translation of mRNAs that encode the different protein subunits that make up L-type voltage-dependent calcium channels (L-VDCC) during long-term synaptic depression (LTD). Specifically, the research will evaluate the translation of the following L-VDCC-associated mRNAs: Cacna1c, Cacna1d, Cacna2d1, Cacna2d2, and Cacna2d3. Assessment on the role of DJ-1 in long-term synaptic depression will be determined by manipulating the expression of DJ-1 virally and by using a mouse model of impaired LTD. To mimic LTD, dissociated hippocampal neurons will be treated with (RS)-3,5-dihydroxyphenylglycine (DHPG). The studies will use a novel protein synthesis assay, surface sensing of translation (SUnSET) coupled with proximity ligation assay (PLA), to determine how DJ-1 controls the translation of L-VDCC-associated mRNAs with temporal and spatial resolution in primary hippocampal neurons. Results from these studies will advance our understanding of the mechanisms underlying hippocampal LTD, which is fundamental to how we encode novel experiences into habituation. Moreover, these studies will establish a novel role for DJ-1 as a regulator of calcium dynamics relevant to learning and memory.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

历史上的黑人学院和大学本科课程（HBCU-UP）通过研究启动奖为历史上的黑人学院和大学的初级和中级职业教师提供支持，他们正在建立新的研究项目或重新定向和重建现有的研究项目。预计该奖项将有助于进一步提高教师的研究能力和效率，并改善家庭机构的研究和教学。这个奖项给北卡罗来纳州AT州立大学提供了一个机会，建立一个分子神经科学实验室，调查机制的学习和记忆。具体来说，该项目旨在研究如何防止L型钙离子通道（允许钙进入神经元的蛋白质）在长期突触抑制（神经元可塑性的一种形式）期间形成。该奖项还包括培养本科生，从而为下一代科学家做出贡献，这些科学家可以为科学创新、创造力和生产力做出贡献。编码学习和记忆，例如区分新旧经验的能力，需要神经元在突触和树突中合成一些蛋白质并抑制其他蛋白质。神经元快速调节其蛋白质水平的能力的一种机制是通过RNA结合蛋白（RBP）。然而，目前对这些RBP的身份、其信使RNA（mRNA）靶点以及这些RBP如何调节其靶点的蛋白质表达的了解还很有限。最近的研究表明，DJ-1，一个未充分表征的RBP，存在于树突和突触，并调节一些树突钙通道的活性。因此，该项目旨在研究DJ-1在调节mRNA翻译中的作用，这些mRNA编码构成L型电压依赖性钙通道（L-VDCC）的不同蛋白亚基。具体而言，该研究将评估以下L-VDCC相关mRNA的翻译：Cacna 1c，Cacna 1d，Cacna 2d 1，Cacna 2d 2和Cacna 2d 3。DJ-1在长期突触抑制中的作用的评估将通过操纵DJ-1的病毒表达和通过使用受损LTD的小鼠模型来确定。为了模拟LTD，将用（RS）-3，5-二羟基苯甘氨酸（DHPG）处理解离的海马神经元。这些研究将使用一种新的蛋白质合成测定方法，表面翻译传感（SUnSET）与邻近连接测定（PLA）相结合，以确定DJ-1如何在原代海马神经元中以时间和空间分辨率控制L-VDCC相关mRNA的翻译。这些研究的结果将促进我们对海马LTD潜在机制的理解，这是我们如何将新体验编码为习惯的基础。此外，这些研究将确立DJ-1作为与学习和记忆相关的钙动力学调节剂的新作用。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。