Research Initiation Award:Transcriptome Analysis of Microtubule Actin Crosslinking Factor 1 (MACF1) Stage-Specific oocytes in Zebrafish

研究启动奖：斑马鱼微管肌动蛋白交联因子 1 (MACF1) 阶段特异性卵母细胞的转录组分析

• 批准号: 2300505
• 负责人: Dondra Bailey
• 金额: $ 29.17万
• 依托单位: Coppin State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-10-01 至 2026-09-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2300505&HistoricalAwards=false
• 关键词: Research; Initiation; Award; Transcriptome; Analysis

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 Coppin State University provides an opportunity to establish a zebrafish laboratory that investigates the mechanisms underlying oogenesis. Specifically, the project aims to examine the function of Microtubule Actin Crosslinking Factor 1 (MACF1) in Balbiani body (Bb) disassembly. With this award, efforts also include the training of graduate, undergraduate, and high school students, thereby contributing to the next generation of scientists who can contribute to scientific innovation, creativity and productivity.The embryonic axis is first determined during oogenesis by polarity cues established in the oocyte, preceding embryonic genome activation. This distinct asymmetry is determined from a vegetally localized structure, called the Balbiani body (Bb). The Bb is conserved in oocytes from vertebrates to invertebrates and is composed of an aggregation of organelles and proteins including RNA-binding proteins. However, the mechanism by which the Bb establishes polarity during oogenesis and disassembles is unknown. There is evidence to demonstrate that a magellan (mgn) mutant causes an egg polarity defect and is characterized by an enlarged Bb and a mis-localized nucleus. The mgn mutant specifically, disrupts the microtubule actin crosslinking factor (macf1a) gene, known to function in other polarized cells. The goal of the proposed study will focus on the earliest stages of oogenesis that result in a mature and viable oocyte for successful fertilization. To address Bb disassembly and the interaction with MACF1a this study will 1) employ Next-Generation Sequencing (NGS) technology to measure differential expression of transcripts in macf1a stage-specific oocytes, and 2) visualize MACF1a protein in combination with the Bb at the ultrastructural level. The proposed project will uncover for the first time the transcriptome of a zebrafish maternal effect mutant (macf1) which may lead to greater insight into the genes controlling polarity while providing high-resolution characterization of the MACF1 protein/Balbiani body.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。该奖项授予了Coppin州立大学，为建立斑马鱼实验室研究卵子发生机制提供了机会。具体而言，该项目旨在研究微管肌动蛋白交联因子1 （MACF1）在巴尔比亚尼体（Bb）分解中的功能。有了这个奖项，还包括研究生、本科生和高中生的培训，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。胚胎轴首先在卵形成过程中由卵母细胞中建立的极性线索决定，在胚胎基因组激活之前。这种明显的不对称是由一种被称为巴尔比亚尼体（Bb）的植物局部结构决定的。Bb在从脊椎动物到无脊椎动物的卵母细胞中都是保守的，它由细胞器和蛋白质（包括rna结合蛋白）的聚集组成。然而，Bb在卵发生和解体过程中建立极性的机制尚不清楚。有证据表明麦哲伦（mgn）突变体导致卵子极性缺陷，其特征是Bb增大和细胞核定位错误。mgn突变体特别破坏了微管肌动蛋白交联因子（macf1a）基因，该基因已知在其他极化细胞中起作用。拟议研究的目标将集中在卵子发生的最早阶段，这将导致成熟和有活力的卵母细胞成功受精。为了研究Bb的分解和与MACF1a的相互作用，本研究将1)采用下一代测序（NGS）技术来测量MACF1a阶段特异性卵母细胞中转录本的差异表达，2)在超微结构水平上可视化MACF1a蛋白与Bb的结合。该项目将首次揭示斑马鱼母体效应突变体（macf1）的转录组，这可能有助于更深入地了解控制极性的基因，同时提供macf1蛋白/Balbiani体的高分辨率特征。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。