Research Initiation Award: FKH-8 Control of Dopamine Signaling

研究启动奖（RIAs）为传统黑人学院和大学（HBCUs）中开始建立研究项目的初级教师提供支持，也为需要重新指导和重建研究项目的职业中期教师提供支持。期望该奖项有助于进一步提高教师的研究能力和效率，改善研究人员所在机构的研究和教学，并使本科生参与研究经验。菲斯克大学（Fisk University）的RIA项目与范德堡大学（Vanderbilt University）合作，测试了转录因子FKH-8调节多巴胺转运蛋白DAT-1上游多巴胺能（DA）神经元功能的假设，这是基于一项令人兴奋的发现，即以前未表征的秀丽隐杆线虫FKH-8基因突变导致多巴胺依赖性麻痹表型。多巴胺能（DA）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特异性基因并关闭某些祖基因或非神经元基因是一个关键步骤，但许多调控机制仍然未知。秀丽线虫是研究DA神经元细胞命运和功能调控的一种高度易处理和强大的模型——它有一个简单、易于观察和操纵的神经系统，同时仍然表达必要的哺乳动物生物合成酶、受体和转运体的同源物。使用这个系统，发现FKH-8，一个以前在秀丽隐杆线虫中没有已知作用的分子，是正常DA功能所必需的。本研究增加了对参与指定和/或维持DA神经元特性的基因调控的理解，并为后续测试基因网络的建议提供了见解和数据，这些建议可以帮助生物工程工作更忠实地概括诱导多能干细胞的DA神经元特征。该项目通过以下目标精确定义了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神经元调节的关键知识。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dat-1的一部分调节DA神经元功能的假设。-通过多管齐下的方法鉴定FKH-8转录因子靶点，包括鉴定新基因，验证FKH-8通过转录调节dat-1和其他DA特异性基因来控制DA功能的假设。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、帕金森病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，菲斯克大学招募了2-3名来自STEM领域代表性不足人群的本科生，将他们整合到由菲斯克硕士到博士桥梁项目学生组成的实验室中。本科生指导包括对遗传学和分子生物学技术的深入实践经验，这些技术是许多生物学学科，学术和专业的基础。更广泛的影响延伸到全州范围内，增加了研究能力和能力，以刺激州内的学术研究，作为NSF刺激竞争性研究的实验计划（EPSCoR）管辖权。

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.