Gene regulatory networks dictating the development and differentiation of the cardiac neural crest
Megan Martik, Professor
Molecular and Cell Biology
Closed. This professor is continuing with Spring 2024 apprentices on this project; no new apprentices needed for Fall 2024.
The neural crest (NC) is a transient stem cell population that emerges during early vertebrate embryogenesis. Characterized by its migratory behavior and multipotency, the NC gives rise to diverse cell types and tissue derivatives including elements of the peripheral nervous system, the craniofacial skeleton, and the cardiovascular system. The NC forms in a series of defined steps—specification, delamination, migration, and differentiation—that are controlled by a gene regulatory network (GRN). Additionally, the NC is partitioned into distinct subpopulations along the anterior-posterior axis corresponding to their final destination in the body plan.
One of the most unique subpopulations is the cardiac neural crest (CdNC), which migrates into the heart and differentiates into portions of the outflow tract, arteries, valves, and cardiomyocytes. We are using the zebrafish as a model to characterize the GRN during CdNC migration and differentiation into cardiomyocytes. Furthermore, we are testing validated genes in the network in the adult injured heart to begin understanding how the CdNC functions during heart regeneration. As a precursor to this work, Dr. Martik published evidence confirming that the CdNC contributes to cardiomyocytes across amniotes and that some canonical NC genes are present during heart regeneration in zebrafish: https://doi.org/10.7554/eLife.47929
Role: The student will be tasked with injecting already-cloned enhancer constructs specific to the migratory or differentiating CdNC into single cell zebrafish embryos. Following injections, embryos will be screened at the correct stage, imaged on a fluorescence microscope, and analyzed. These experiments will allow the student to gain an understanding of zebrafish transgenesis and enhancer biology. Alongside this, the student will also clone new enhancer sequences into a vector backbone amenable to transgenesis. This will involve Gibson assembly, bacterial work, and DNA sequence analysis. These new plasmids will then be injected into the single cell embryo to validate their expression at specific stages. Alongside this, the student will be tasked with reading and discussing with their mentor literature in the field to gain an understanding of neural crest and enhancer biology, genetic modifications in zebrafish, and gene regulatory networks. If the student can dedicate a lot of time to research, there are opportunities to learn additional experimental techniques such as hybridization chain reaction (HCR), confocal microscopy, and more.
Students will interact directly with their mentor, Luke Lyons, for day-to-day tasks and will meet with Dr. Martik weekly to discuss research progress.
Qualifications: We are looking for a highly motivated and focused student interested in developmental biology and gene regulation. Familiarity with molecular biology (restriction enzymes, PCR, gel electrophoresis, bacterial transformation, etc) is essential and an understanding of basic genetics is desirable. Previous wet lab experience is not required, but helpful. Ideally, this student would have a genuine interest and curiosity in the questions being asked, and a cogent understanding for how URAP will aid their professional development. It is paramount to that research is taken seriously and that the student can dedicate a substantial amount of time to the project.
MCB Major – Required
Bio 1A – Required
Biochemistry 102 or equivalent (e.g., C110) – Highly desirable but not essential
Genetics 104 or 140 – Highly desirable but not essential
Developmental Biology 141 – Desirable but not essential
Availability over the summer for research – Desirable but not essential
Students from underrepresented backgrounds in science are encouraged to apply.
Day-to-day supervisor for this project: Luke Lyons, Staff Researcher
Hours: 12 or more hours
Related website: www.martiklab.org
Biological & Health Sciences