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Effect of DSK gastro-peptide on feeding behaviors in fruit flies
- 1. The effect of the DSK gastro-peptide on feeding behaviors in Drosophila Melanogaster By: Christina Mary Joseph Mentor: Dr. Monica Dus, Ph.D Skirball Institute of BioMedicine, NYU
- 2. Laying down the genetics
- 3. Laying Down the genetics
- 4. Known Behaviors 1.000250 267 2.112347 667 Sated starved Amounteaten Condition Food ingestion assay of wildtype flies demonstrate the effect of satiety on ingestion rate
- 5. Known Behaviors 26.27422 605 87.88537 29 Sated starved Percentoffliesthatate Condition of fly W1118cs food availibilty assayFood availability assay of wildtype flies demonstrate the effect of satiety on commencement of eating under low availability circumstances
- 6. Experiments: PER Assay Proboscis Extension Reflex Assay. Aids in determining feeding behaviors
- 7. Experiments: Ingestion Assay The ingestion assay aids in determining quantity of food intake.
- 8. Experiments: Food availability assay The food availability assay aids in determining ingestion initiation behaviors
- 9. What we found Figure 1: Percentage of flies that extended their proboscis in both genotypes. Figure 2: Trends of proboscis extension (TRPA1/DSK- Activated DSK neurons)
- 10. What we found The amount of food eaten by the flies (1.DSK/KG80- down regulation of DSK 2.DSK/NaChBac- up regulation of DSK) 0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 DSK/+ DSK> kir2.1, tubulingal80ts DsK>NAchBacamounteaten Genotypes Ingestion assay
- 11. What we found Food availibility results (1.DSK/KG80- down regulation of DSK 2.DSK/NaChBac - up regulation of DSK) 0 10 20 30 40 50 60 DSK/+ DSK> kir2.1, tubulingal80ts 102039 CCKLR-17D3 Percentageoffliesthatate Genotypes Food availibility assay: starved
- 12. Conclusion • The food availability assay showed that starved flies with increased levels of DSK tend to choose not to eat but it did not show starved flies with decreased levels of DSK ate more • The food ingestion data showed amount eaten by the flies with less DSK ate greater portions than the control while flies with more DSK ate less portions than that of the control • The PER assay showed that silencing of the DSK neurons led to increased proboscis extension
- 13. Discussion • The data shows strong preliminary promise and a second round of experiments replicating the data shown here will allow us to move into a mammalian model; mice. • If the data is replicated in the mammalian model, than eating disorders can be prevented by genetic manipulation also thereby preventing long term health defects such as heart disease, kidney failure and even cancer.
- 14. THANK YOU
Editor's Notes
- f genetic modification of DNA is done, the result is called "knockdown organism." If the change in gene expression is caused by an oligonucleotide binding to an mRNA or temporarily binding to a gene, this leads to a temporary change in gene expression that does not modify the chromosomal DNA, and the result is referred to as a "transient knockdown".[1] In a transient knockdown, the binding of this oligonucleotide to the active gene or its transcripts causes decreased expression through a variety of processes. Binding can occur either through the blocking of transcription (in the case of gene-binding), the degradation of the mRNA transcript (e.g. by small interfering RNA (siRNA) or RNase-H dependent antisense), or through the blocking of either mRNA translation, pre-mRNA splicing sites, or nuclease cleavage sites used for maturation of other functional RNAs, including miRNA(e.g. by Morpholino oligos or other RNase-H independent antisense).[1][2] The most direct use of transient knockdowns is for learning about a gene that has been sequenced, but has an unknown or incompletely known function. This experimental approach is known as reverse genetics. Researchers draw inferences from how the knockdown differs from individuals in which the gene of interest is operational. Transient knockdowns are often used in developmental biology because oligos can be injected into single-celled zygotes and will be present in the daughter cells of the injected cell throughembryonic development.[3]