Research Initiation Award: FKH-8 Control of Dopamine Signaling

研究启动奖(RIA)为开始建立研究计划的历史黑人学院和大学(HBCU)的初级教师提供支持，以及为需要重新指导和重新建立研究计划的职业生涯中期教师提供支持。Fisk大学的RIA项目与范德比尔特大学合作，测试了转录因子FKH-8调节多巴胺转运体DAT-1上游的多巴胺(DA)神经元功能的假设，基于一项令人兴奋的发现，即以前未描述的线虫FKH-8基因的突变会导致多巴胺依赖的瘫痪表型。多巴胺(DA)能神经元调节驱动运动和认知行为的许多方面的关键信号通路。在DA神经元中，开启正确的DA特异性基因和关闭某些前体或非神经元基因是关键的一步，但许多调控机制仍不清楚。线虫是研究DA神经元细胞命运和功能调控的一个高度易处理和强大的模型--它有一个简单、容易观察和操作的神经系统，同时仍然表达哺乳动物必要的生物合成酶、受体和转运蛋白的同源物。使用这个系统，发现FKH-8，一个以前在线虫中没有已知作用的分子，是正常DA功能所必需的。这项研究增加了对指定和/或维持DA神经元特性所涉及的基因调控的理解，并为后续测试遗传网络的提案提供了洞察力和数据，有助于生物工程努力更真实地概括诱导多能干细胞的DA神经元规格。该项目通过以下目标准确地定义了FKH-8在多巴胺信号通路中的位置，总体目标是获得适用于许多物种的DA神经元调控的关键知识。该项目的两个目标是：-通过进行遗传、药理学和生化分析，验证FKH-8作为同一通路的一部分调节DA神经元功能的假设。-测试FKH-8通过转录调控dat-1和其他DA特异性基因来控制DA功能的假设，通过多管齐下的方法识别FKH-8转录因子的靶标，包括新基因的识别。该项目在发育生物学方面的基础研究有可能为未来与成瘾、瘫痪、帕金森氏症和精神分裂症相关的研究和技术做出贡献。此外，通过该项目，FISK大学吸收了2-3名STEM领域代表性不足的本科生，将他们整合到由FISK硕士到博士桥项目学生组成的实验室中。本科生辅导包括深入的实践经验，遗传学和分子生物学技术的基础，许多生物学学科，学术和专业。更广泛的影响延伸到全州范围内，作为一项刺激竞争性研究(EPSCoR)管辖权的NSF实验计划，提高了研究能力和刺激州内学术研究的能力。

Research Initiation Awards (RIAs) provide support for junior faculty at Historically Black Colleges and Universities (HBCUs) who are starting to build a research program, as well as for mid-career faculty who need to re-direct and re-build a research program. It is expected that the award helps to further the faculty member's research capability and effectiveness, improves research and teaching at the researcher's home institution, and involves undergraduate students in research experiences.The RIA project at Fisk University, in collaboration with Vanderbilt University, tests the hypothesis that the transcription factor FKH-8 regulates dopaminergic (DA) neuron function upstream of the dopamine transporter, DAT-1, based on an exciting discovery that mutation of the previously uncharacterized C. elegans FKH-8 gene leads to a dopamine-dependent paralysis phenotype. Dopaminergic (DA) neurons regulate critical signaling pathways that drive many facets of motor and cognitive behavior. In DA neurons, switching on the right DA-specific genes and switching off certain progenitor or non-neuronal genes is a critical step, but much of the regulatory machinery is still unknown. The nematode C. elegans is a highly tractable and powerful model for studying regulation of DA neuron cell fate and function-- it has a simple, readily observed and manipulated nervous system, while still expressing homologs of the essential mammalian biosynthetic enzymes, receptors, and transporters. Using this system, the discovery that FKH-8, a molecule with no previously known role in C. elegans, is necessary for normal DA function. This research leads to increased understanding of gene regulation involved in specifying and/or maintaining DA neuron identity, and provide insight and data for subsequent proposals testing genetic networks that could help bioengineering efforts to more faithfully recapitulate DA neuron specification from induced pluripotent stem cells.The project precisely defines the position of FKH-8 in the dopamine signaling pathway through the following objectives, with the overall goal of gaining key knowledge of DA neuron regulation applicable across many species. The project's two objectives are to:-Test the hypothesis that FKH-8 regulates DA neuron function as part of the samepathway as dat-1 by performing genetic, pharmacological, and biochemical assays. -Test the hypothesis that FKH-8 controls DA function through transcriptional regulationof dat-1 and other DA-specific genes by a multi-pronged approach to identify targets of the FKH-8transcription factor to include identification of novel genes. The basic research from this project in developmental biology has the potential to contribute to future studies and technologies connected to addiction, paralysis, Parkinson's disease and schizophrenia. In addition, through this project, Fisk University involves 2-3 undergraduates from populations underrepresented in STEM fields, integrates them into the lab which is staffed by Fisk Masters-to-PhD Bridge Program students. Undergraduate mentoring involves in-depth hands-on experience with genetics and molecular biology techniques fundamental to many biological disciplines, academic and professional. The broader impact stretches statewide in increasing research capability and capacity to stimulate academic research within the state as an NSF Experimental Program to Stimulate Competitive Research (EPSCoR) jurisdiction.