Characterizing Tg[GT1: HTG] Line of Zebrafish
Mentor 1
Henry Tomasiewicz
Location
Union Wisconsin Room
Start Date
24-4-2015 10:30 AM
End Date
24-4-2015 11:45 AM
Description
The Tg[GT1: HGT] (GT1) line of zebrafish, which was created using a gene trap approach, expresses Human-tau Green Fluorescent Protein (HTG) from the promoter of an unknown. HTG serves as a rough indicator to show when the GT1 gene is normally expressed. In GT1, HTG is expressed only in the ovaries and in embryos from GT1+ females during the first 30 hours post-fertilization. During the first 4-5 hours of life, until the midblastula transition, when the paternal genes start to become active, zebrafish rely solely on maternal expressed genes. During the midblastula transition, the zygotic (embryonic) genome becomes active, and sometime after this, GT1 gene becomes inactivated and HTG expression ceases. When GT1 becomes activated again in the developing ovaries is unknown and of interest to us. Our goal was to determine when GT1 turns back on during zebrafish development. To do this, we first needed to spawn a GT1 female with a sheer male fish. The offspring were then examined under a fluorescent microscope to reveal the expression of HTG. Then, these offspring were examined at weekly intervals to determine when it appears again. We also used polymerase- chain reaction (PCR) to identify GT1 gene in male fish. Present findings show that 30+ day- old zebrafish have been expressing HTG. When the time line for HTG expression is finalized and the gene is identified, a better understanding of zebrafish development and their genetic makeup will be available. Because zebrafish are largely used as models for human disease, defining what these genes are and when they impact the zebrafish will be helpful for researchers in the biomedical field.
Characterizing Tg[GT1: HTG] Line of Zebrafish
Union Wisconsin Room
The Tg[GT1: HGT] (GT1) line of zebrafish, which was created using a gene trap approach, expresses Human-tau Green Fluorescent Protein (HTG) from the promoter of an unknown. HTG serves as a rough indicator to show when the GT1 gene is normally expressed. In GT1, HTG is expressed only in the ovaries and in embryos from GT1+ females during the first 30 hours post-fertilization. During the first 4-5 hours of life, until the midblastula transition, when the paternal genes start to become active, zebrafish rely solely on maternal expressed genes. During the midblastula transition, the zygotic (embryonic) genome becomes active, and sometime after this, GT1 gene becomes inactivated and HTG expression ceases. When GT1 becomes activated again in the developing ovaries is unknown and of interest to us. Our goal was to determine when GT1 turns back on during zebrafish development. To do this, we first needed to spawn a GT1 female with a sheer male fish. The offspring were then examined under a fluorescent microscope to reveal the expression of HTG. Then, these offspring were examined at weekly intervals to determine when it appears again. We also used polymerase- chain reaction (PCR) to identify GT1 gene in male fish. Present findings show that 30+ day- old zebrafish have been expressing HTG. When the time line for HTG expression is finalized and the gene is identified, a better understanding of zebrafish development and their genetic makeup will be available. Because zebrafish are largely used as models for human disease, defining what these genes are and when they impact the zebrafish will be helpful for researchers in the biomedical field.
