Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology
Yuriyah Reed-Harris - Oregon State University
Co-Author(s): John Gamble and Julie Greenwood, Oregon State University, Corvallis, OR
Glioblastoma cells retain the exceptional ability to permeate the brain and migrate long distances from the original tumor. This migration makes traditional treatments such as chemotherapy and radiation therapy ineffective in most cases and complete surgical resection virtually impossible. Little is known about how the glioblastoma cells form attachments and migrate along the basement membrane in the complex microenvironment of the brain. We are using Fli-fish, which are called Tg(fli1:eGFP)y1 zebrafish. This specific notation is used for realization that the transgene or gene added in was eGFP (enhanced green fluorescent protein) with the fli1 promoter; Y1 is the specific allele. Fli-fish use is key because they express green florescent protein (GFP) which labels the blood vessels. It is thought that laminins mediate the attachment, migration, and organization of cells into tissues during embryonic development by interacting with both cellular and extracellular matrix components. This is a common thought because the endothelial cells organize the extracellular matrix as well as bind to the laminin α-5. It is known that glioblastoma binds to laminin α-5 via the integrin binding site, so in order to investigate the mechanisms by which the glioblastoma cells migrate and proliferate, the integrin binding site of laminin α-5 was knocked down by injecting a splice blocking morpholino into a single cell fertilized zebrafish embryo. Then human glioblastoma cells, dyed with CM-dil, were injected into the zebrafish brain two days post fertilization (dpf).
We’re testing the hypothesis that by knocking down laminin α-5 we will understand the role and mechanism by which the laminin alpha 5 is regulating invasion. Time lapse microscopy and 4D imaging, conducted 3-4 dpf, were used to determine cell morphology and the mechanisms of movement through increased magnification. Results measuring the tumor cell velocity, directionality, and proximity to blood vessels will be presented. Future research involves precise measurements in order to assure the maximum consistency of embryo quality, quantification of the exact volume of cells being injected into every fish, and exploration of the Image J software for measurements.
Funder Acknowledgement(s): This research was supported by a grant from the NSF awarded to, Julie Greenwood Ph.D, Associate Dean of the College of Science, Oregon State University, Corvallis, OR. This study was propelled thanks to the resources of the SURE Science program, directed by Julie Greenwood Ph.D, Associate Dean of the College of Science, Oregon State University, Corvallis, OR. The research opportunity as a whole was presented and made possible by the STEM Leaders program lead by Dr. Kevin Ahern, Director of Undergraduate Research, Oregon State University, Corvallis, OR.
Faculty Advisor: Julie Greenwood,