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

历史上的黑人学院和大学本科项目（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期职业教师提供支持，他们正在建立新的研究项目或重新指导和重建现有的研究项目。期望该奖项有助于进一步提高教师的研究能力和效率，并改善本国机构的研究和教学。该奖项授予北卡罗来纳农工州立大学，为建立一个分子神经科学实验室提供了一个机会，该实验室将研究学习和记忆的潜在机制。具体来说，该项目旨在研究l型钙离子通道（允许钙进入神经元的蛋白质）是如何在长期突触抑制（神经元可塑性的一种形式）期间被阻止形成的。该奖项还包括培养本科生，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，比如区分新旧经验的能力，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。神经元快速调节其蛋白质水平的一种机制是通过rna结合蛋白（rbp）。然而，目前关于这些rbp的身份，它们的信使RNA （mRNA）靶标，以及这些rbp如何调节其靶标的蛋白质表达的知识知之甚少。最近的研究表明，DJ-1是一种未被充分表征的RBP，它存在于树突和突触中，并调节一些树突钙通道的活性。因此，本项目旨在研究DJ-1在长期突触抑制（LTD）期间调节组成l型电压依赖性钙通道（L-VDCC）的不同蛋白质亚基编码mrna翻译中的作用。具体而言，本研究将评估以下l - vdcc相关mrna的翻译：Cacna1c、Cacna1d、Cacna2d1、Cacna2d2和Cacna2d3。DJ-1在长期突触抑制中的作用将通过病毒控制DJ-1的表达和使用受损的小鼠LTD模型来确定。为了模拟LTD，分离的海马神经元将用(RS)-3,5-二羟基苯基甘氨酸（DHPG）处理。这些研究将使用一种新的蛋白质合成试验，表面翻译传感（SUnSET）结合接近结扎试验（PLA），以确定DJ-1如何通过时间和空间分辨率控制初级海马神经元中l - vdcc相关mrna的翻译。这些研究的结果将促进我们对海马体有限责任的机制的理解，这是我们如何将新经历编码为习惯的基础。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。该奖项反映了美国国家科学基金会的法定使命，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。

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.