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

通过研究启动奖的历史黑人学院和大学本科课程(HBCU-UP)为历史黑人学院和大学的初级和职业生涯中期教师提供支持，这些教师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。预计该奖项有助于进一步提高教师的研究能力和有效性，并改善所在机构的研究和教学。这项授予北卡罗来纳A&T州立大学的奖项提供了一个建立分子神经科学实验室的机会，该实验室研究学习和记忆的潜在机制。具体地说，该项目旨在研究L型钙离子通道是如何在长期突触抑制期间阻止钙进入神经元的蛋白质的，突触抑制是神经元可塑性的一种形式。获得该奖项的努力还包括培养本科生，从而为下一代科学家做出贡献，他们可以为科学创新、创造力和生产力做出贡献。编码学习和记忆的能力，如区分新旧经历的能力，需要神经元合成一些蛋白质，并抑制突触和树突中的其他蛋白质。神经元快速调节蛋白质水平的一个潜在机制是通过RNA结合蛋白(RBPs)。然而，目前对这些限制性商业惯例的身份、它们的信使RNA(MRNA)靶标以及这些限制性商业惯例如何调控其靶标的蛋白质表达的了解很少。最近的研究表明，DJ-1是一种功能不强的RBP，存在于树突和突触中，调节一些树突状钙通道的活动。因此，本研究旨在探讨DJ-1在长期突触抑制(LTD)过程中对L型电压依赖性钙通道(L-VDCC)不同蛋白质亚基表达的调控作用。具体地说，本研究将评估以下与L VDCC相关的mRNAs的翻译：CACNA1C、CacNA1d、Cacna 2d1、Cacna 2d2和Cacna 2d3。对DJ-1在长期突触抑制中的作用的评估将通过病毒操纵DJ-1的表达和使用受损有限公司的小鼠模型来确定。为了模拟LTD，分离的海马神经元将被(RS)-3，5-二羟基苯甘氨酸(DHPG)处理。本研究将使用一种新的蛋白质合成实验，即翻译表面感觉(日落)与邻近连接分析(PLA)相结合，以确定DJ-1如何控制原代海马神经元中具有时间和空间分辨率的L-VDCC相关mRNAs的翻译。这些研究的结果将促进我们对海马体LTD潜在机制的理解，这是我们如何将新体验编码为习惯化的基础。此外，这些研究将为DJ-1建立一个与学习和记忆相关的钙动力学调节器的新角色。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.