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

Coppin State University, Baltimore MD

Investigators

Abstract

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.