Investigating The Role Of Dnc-2 And Dli-1 On Ampa Receptor Mediated Behaviors In C. Elegans

γ-amino-3-hydroxy-5methylisoxazole-4-proprionic acid (AMPA) receptors (AMPARs) are protein complexes involved in excitatory neurotransmission. AMPARs are tetrameric structures consisting of the pairings of GluA1, GluA2, GluA3, and GluA4 subunits. Dctn2 and Dync1li1 were found to interact with GluA2 in an immunoprecipitation screen that was performed on postnatal day 14 rat brains. DNC-2 and DLI-1 were identified as the C. elegans homologs of Dctn2 and Dync1li1, respectively, and are components of motor proteins that transport cargo throughout the cell. The goal of this project was to determine whether knocking down DNC-2 and DLI-1 by RNAi affects AMPA Receptor-mediated behaviors, including mechanosensation and chemosensation, in C. elegans

Analysis of FBP1 Gene Expression in UV Light-exposed Tetrahymena thermophila Cultures

In this study, Tetrahymena thermophila cultures were exposed to an acute treatment of UV light and expression of the FBP1 gene was examined. The FBP1 gene encodes Fructose- 1,6-biphosphatase which plays an important role in the process of gluconeogenesis. It was hypothesized that exposure of Tetrahymena to UV light would cause an upregulation in DNA repair enzymes, thereby causing these cells to require higher levels of glucose. The higher demand for glucose would then result in the upregulation of gluconeogenesis and FBP1. Following acute exposure of Tetrahymena cultures to UV light, the expression of FBP1 was analyzed using reverse transcription and semi-quantitative PCR. It was predicted that the levels of FBP1 would be upregulated following acute UV light treatment

UV Light Exposure and Tetrahymena thermophilia: Expression of the PHR2 Gene

The PHR2 (photolyase repair) gene functions in the repair of DNA in Tetrahymena thermophila. It was hypothesized that when Tetrahymena cultures are exposed to UV light, increased expression of PHR2 would occur in response to the damage incurred. UV light has been known to cause the formation of pyrimidine dimers and other disruptions in DNA. Through two rounds of experimentation, the acute impact of UV light exposure was monitored. The cultures underwent RNA extraction, reverse transcription, and semi-quantitative PCR analysis. We predicted that UV exposure would result in the increased expression of PHR2

Put Your Phone Down: Effects of Cellular Radiation on Drosophila melanogaster

In this project, we aimed to answer the question: Does cellular radiation affect offspring production and expression of the Rad51 gene in the organism Drosophila melanogaster? It was hypothesized that if Drosophila larvae were exposed to cellular radiation, the total offspring production would decrease, along with a corresponding increase in Rad51 expression. The Rad51 protein is crucial to the propagation of strand invasion and exchange steps in homologous recombination, resulting in the repair of double stranded DNA breaks. To test our hypothesis, we exposed the experimental group of Drosophila larvae to doses of cellular radiation emitted from an iPhone 6 or 7. Exposure to cellular radiation occurred for 6-minute durations, twice daily, for 3 consecutive days. At the end of the treatment period, RNA extraction from larvae and complimentary Reverse Transcription Polymerase Chain Reaction (RT-PCR) was performed on both control and treatment groups to measure the expression of Rad51. Further analyses were performed to look at the fertility of Drosophila larvae that were exposed to cellular radiation. Due to prior research on cellular radiation exposure and its connection to DNA and sperm damage, it was predicted that expression of the Rad51 gene would show an increase in our treatment groups, along with a corresponding decrease in reproductive ability

Effects of the Everyday Toxin, Titanium Dioxide, on Drosophila melanogaster Nervous System Development

Titanium dioxide (TiO2) is a widely used compound found in everything from food packaging to sunscreens. When ingested, TiO2 is readily transported across membranes and efficiently stored within cells. Previous studies showed that exposure to TiO2 results in underdeveloped nervous systems. For our study, we attempted to answer the following question: Will exposing Drosophila melanogaster larvae to TiO2 affect expression of the Neur gene and development of the central nervous system? The Neur gene is crucial during the celldetermination stage of development as its encoded protein helps specify neuroblast development and aids in nervous system and sensory organ development. It was hypothesized that expression of the Neur gene would decrease in Drosophila larvae exposed to TiO2 and that nervous system development would be abnormal compared to control larvae. To test this hypothesis, Drosophila larvae were randomly assigned to either a control group, which was cultured under ideal conditions, or a treatment group, which was exposed to a non-lethal concentration of TiO2. Following exposure, RNA extraction and Reverse Transcription Polymerase Chain Reaction (RT-PCR) was conducted to analyze expression of Neur. To quantify nervous system development, Drosophila larvae were subjected to a touch-response assay. Because TiO2 likely interferes with neural development, it was predicted that Drosophila larvae would show decreased expression of Neur and that they would respond poorly to a mechanical touch-response assay

Effect of Nitrate Pollution on Tetrahymena thermophila

The question under investigation in this project was: Does an elevated concentration of nitrates have a negative eect on cell growth and expression of the CDC16 gene in the organism Tetrahymena thermophila? Exposure of Tetrahymena cultures to nitrates mimics the exposure of these organisms to nitrate pollution in the environment, which can occur due to septic tank leakage, nitrogen-rich fertilizer run o, and agricultural processes. The EPA standard for nitrate concentrations in drinking water is a maximum of 10ppm. In our experiment, Tetrahymena cultures were treated with media that contained 30ppm of nitrates for one week. The 30ppm nitrate concentration has been proven acceptable for freshwater fish; therefore, we predicted that it would have more of an impact on our Tetrahymena cells which are far less complex. The CDC16 gene encodes a Cell Division Cycle protein that contains an anaphase-promoting complex (APC). This APC is involved in Cyclin degradation and promotes the movement of the cell out of mitosis. During the oneweek exposure to 30ppm nitrates, the growth rate of Tetrahymena cultures was determined using a hemocytometer. Furthermore, expression of the CDC16 gene was analyzed using semi-quantitative RT-PCR following the weeklong exposure. We predicted that a high concentration of nitrates would decrease the growth rate and expression of CDC16 in our Tetrahymena cultures

Effects of Sulfur Dioxide on abdA Gene Expression and Larval Development in Drosophila melanogaster

In this experiment, we tested the effects sulfur dioxide exposure had on abd-A gene expression and larval development in Drosophila melanogaster. Previous studies showed that sulfur dioxide exposure inhibits development of gonads in adult flies and increases development time. We hypothesized that expression of abd-A would decrease in developing Drosophila exposed to sulfur dioxide. The abd-A gene is in the Hox gene family, which functions in the development of body segmentation. The abd-A gene is specific to the abdominal region of the fly, and is also important to the development of gonads and fat bodies. To test our hypothesis, we randomly selected three sets of mating pairs and allowed them to lay eggs in larval culture dishes. The experimental group of larvae was subjected to sulfur dioxide continuously for seven days, while the control group was not exposed to sulfur dioxide, but kept under the exact same conditions. After the exposure, fly larvae were extracted from their culture dishes, sorted by developmental stage, and counted. RNA was then extracted from the larvae after counting. Reverse-Transcription Polymerase Chain Reaction (RT-PCR) was performed to determine abd-A expression in experimental and control groups. Due to sulfur dioxide having detrimental effects on gonad and larval development, it was predicted that, in the presence of sulfur dioxide, abd-A gene expression would decrease, and developmental time course of larval development would be delayed

The Effects of Warming Temperatures on Rad51 Expression in Tetrahymena thermophila

Our experiment explored whether raised temperatures affected expression of the Rad51 gene in Tetrahymena thermophila. Our hypothesis was that raised temperatures would result in a decrease in expression of the Rad51 gene. The protein product of this gene functions in double stranded DNA repair. We chose this experiment to see if the rising average temperatures around the world would affect expression of genes involved in DNA repair. Our test samples were kept in a growth medium at 37C while our control samples were kept in a growth medium at 24C. Afterwards, the transcribed mRNA was reverse transcribed into DNA and primers were used to amplify the Rad51 gene. Semi-quantitative PCR was then used to analyze expression of the Rad51 gene. Our prediction was that higher temperatures would decrease the expression of the Rad51 gene

The Effect of Basic Conditions on CDK1 Expression in Tetrahymena thermophila

The goal of this project was to discover whether expression of the CDK1 gene in Tetrahymena thermophila, as well as their growth, are aected by changing the pH of the growth media. We hypothesized that expression of the CDK1 gene would decrease significantly when grown in basic media. The CDK1 gene encodes the protein Cyclin-Dependent Kinase, which functions to advance cells through various stages of the cell cycle. To test our hypothesis, Tetrahymena cultures were randomly assigned to either the control group, which was grown in media with a neutral pH, or a treatment group, which was grown in media with a basic pH of 8.5. Previous experiments showed that Tetrahymena were able to survive in pH levels up to 9.0, although their optimal survival was at a neutral pH. The control and experimental cultures were maintained for one week before being processed. Following the weeklong incubation, RNA extraction, reverse transcription, and PCR were conducted on the control and experimental groups to measure expression of CDK1. In order to look at growth rate, we used a hemocytometer to perform cell counts throughout the weeklong incubation. Because CDKs are a major component of the cell cycle, we predicted that expression of the CDK1 gene would decrease, and that growth rate would also decrease, in Tetrahymena cells cultured in a basic environment

Effects of UVB light on Tfb5 gene expression in developing Drosophila melanogaster

UVB light is known to cause DNA damage that results in gene mutations. Most regions of DNA damage are corrected using Nucleotide Excision Repair mechanisms (NER). In Drosophila melanogaster, the Tfb5 gene codes for an essential subunit of a protein involved in NER-mediated DNA repair. In this experiment, we attempted to answer the question: will overexpression of the Tfb5 gene in Drosophila compensate for the damage done to DNA by UVB light exposure? In testing this hypothesis, we exposed our Drosophila larvae cultures to 7-minute periods of UVB light once during each stage of larval development. Our control group grew in conditions without UVB light. Larvae counts in both the experimental and control groups served as our quantitative analysis of survivability. Reverse Transcription Polymerase Chair Reaction (RT-PCR) was performed on the extracted larval RNA to determine the level of expressed Tfb5 gene in our control and experimental groups. Because our Drosophila larvae were exposed to UVB light early on in development, we predicted there would be an overexpression of the Tfb5 gene in the experimental group, along with similar numbers of living larvae in both groups