GREAT Day

2024 Virtual GREAT Day Program

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  • Aguilar, Cesar
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Poster

Loss of Function Mutation for tp53 does not Rescue the chaf1b^nt2 Small-eye Phenotype in Danio rerio

Alexander Parks

FACULTY SPONSOR: Travis Bailey, Biology

In Zebrafish, the chromosome assembly factor 1b (chaf1b) gene is in part responsible for the development of the eye. In homozygous chaf1bt24412 mutants retinal cell death is promoted through cell-death promoting activity of the gene, tumor suppressor protein p53 (tp53), resulting in a small-eye phenotype. Another allele chaf1bnt2, was found to also result in the small-eye phenotype when in a homozygous state. We found that knockdown of Tp53 protein via morpholino antisense oligonucleotide injection of 1-2 cell stage embryos failed to rescue retinal cell death of chaf1bnt2 homozygous mutants as detected by TUNEL labeling. Because morpholinos may fail to fully inhibit target gene function we crossed carrier fish heterozygous for both the nt2 and a cell-death induction deficient allele of tp53 (zdf1) and compared double homozygous mutants to siblings which had chaf1b mutant homozygosity and functional tp53. Restriction fragment length polymorphism analysis was used to verify zygosity of the nt2 and zdf1 alleles. We found that loss of tp53 function failed to rescue the chaf1bnt2 small-eye phenotype.

Poster

Analysis of Her4.1 and Ascl1a in gef Mutants

Tessa Beiter , Brynn Johnson , Lin Kai Ye

FACULTY SPONSOR: Travis Bailey, Biology

Zebrafish make an ideal model organism for studying the development of the retina. The mutant good effort (gef) zebrafish have smaller eyes compared to the wild-type zebrafish embryos at 2 days post-fertilization. This is due to retinal degradation because of the lack of a functioning Chaf1b protein. The Chaf1b protein-coding region is disrupted due to the deletion of its coding intronic DNA which causes an exon to be lost. Chaf1b is important to the cell because it makes up one of the three parts of the chromosome assembly factor 1 (CAF-1). CAF-1 functions to regulate chromatin and load histones onto DNA, but cannot function without Chaf1b. While it has been hypothesized that Tp53-mediated apoptosis is responsible for the gef mutants’ small eyes, tp53-morphants and homozygous gef double mutants have been studied and the cell death seen in gef mutants is not correlated with Tp53 activity. Retinal regeneration signaling pathway members, Her4.1 and Ascl1a, promote retinal cell fate specification. Histone deacetylases selectively regulate her4.1 and ascl1a during retinal development, making these genes possible affected genes of the gef phenotype. These two genes were visualized in gef and wild-type zebrafish embryos via the in situ hybridization process.

Poster

Single Presentation/Performance

Identifying Non-Regenerating Zebrafish Mutants

Austin Feasley , Arianna Falletta

FACULTY SPONSOR: Travis Bailey, Biology

Zebrafish are a useful model organism in studying developmental biology. One topic of interest is their ability to regenerate tissues, including retinal cells. Uncovering genetic pathways for this disease could be applicable to humans, as humans and zebrafish have numerous analogous genes. Scientists use mutants with non-functioning genes to elucidate where genes lie in a pathway. We studied mutants, which are suspected to lack the regenerative abilities of wild type zebrafish; the specific gene is unknown. Our goal was to confirm our family of fish as regenerative mutants. First, they were dark adapted and then placed in intense light for three days. We examined the retinas for regeneration under a fluorescence microscope. If they are regenerating they should be brightly expressing GFP (protein that glows under fluorescent light), but fail to express bright GFP if they are not. The eyes were surgically removed and cryosectioned. From here, the slides will be further examined under a confocal microscope in order to see if the antibodies bind to lower numbers of retinal cells in our mutant line compared with normal fish. We then plan on repeating this experiment for the other zebrafish in the same line as these parent zebrafish.

Poster

Synchronous Q&A: Biology Posters & Talks

Lydia Fregosi , Jessica Palmeri

FACULTY SPONSORS: Mackenzie Gerringer, Biology
Varuni Jamburuthugoda, Biology
Sara Burch, Biology
Travis Bailey, Biology

Join us on Zoom on Friday, April 30th at 2:30 pm for a synchronous discussion to celebrate undergraduate research in Biology at this year's GREAT Day. We invite you to view the Biology talks and posters ahead of time and bring your questions for all presenters to this live session. See you there! Q&A Moderators: Lydia Fregosi ('21) and Jess Palmeri ('21).

Manipulating Cancer Stem Cells: Their Role in Cancer Progression Turned to Therapeutics

Sakura Hamazaki

FACULTY SPONSOR: Travis Bailey, Biology

Breast cancer is the most prevalent disease in the world. This popularity has pushed scientists to make advances in early detection methods, treatments, and public awareness to help alleviate the fatality of this disease; however, the number of new cases per year continue to rise with 250,520 cases reported to the Center of Disease Control in 2017 compared to 196,628 new cases recorded in 1999. This high frequency is commonly attributed to the physiology of the breast, consisting of tissue cells of heterogenetic nature, as this provides an opportunistic microenvironment prone to cancer metastasis. Cancer stem cells have drawn the attention of researchers due to their unique ability to retain characteristics found in stem cells: the ability to self-renew, influence neighboring cells, and be unaffected by traditional targeted therapeutics like chemotherapy (Clarke, et.al). Due to these characteristics, if cancer stem cells are not properly targeted, the disease is likely to reoccur. Interactions between aldehyde dehydrogenase and cytokines IL-6 and CXCL7 provide a foundational example of the relationship between cancer stem cells and the surrounding tissue, promoting cancer progression and possessing the potential to be a biomarker for cancer stem cell identification.

Video (pre-recorded)