Date of Award

Spring 4-25-2016

Degree Type

Thesis

Degree Name

Bachelor of Science

Department

Biology

First Advisor

Dr. Matthew DeGennaro

Abstract

Aedes aegypti is a vector for the majority of mosquito-borne illnesses. The capability of this vector depends on the need for female mosquitoes to feed on human blood to nourish their eggs. The female mosquitoes ability to find human hosts is dependent on both odor detection and changes in neural circuits in the brain. In A. aegypti, Juvenile Hormone is stimulated in vitro by the neuropeptides allatotropins (AT) and inhibited by allatostatins (AS), which are secreted from the brain and target the corpora allata. Allatotropin in A. aegypti has been shown to stimulate JH synthesis in the corpora allata in ex vivo studies. Due to its connection with Juvenile Hormone synthesis, allatotropin is likely an essential factor contributing to reproductive maturation and the gonotrophic cycle in A. aegypti. Although addition of allatotropin to the corpora allata ex vivo has been shown to stimulate synthesis of JH, its role in vivo has not been described. Here I use CRISPR/Cas9 genome editing to excise the allatotropin neuropeptide and make two null mutant lines. A deletion formed by non-homologous end joining, ATdel, and a 50bp insertion of a docking strain using homology dependent repair, ATattP. These mutants show a significant divergence from expected Mendelian ratios (ATdel p=2.05x10- 16, df=2 and ATattP p=0.003, df=2). The genotypic divergence from expected genetic ratios seen in the offspring of two heterozygous progenitors in both mutant lines shows lethality of the mutation suggesting that the Allatotropin neuropeptide plays a significant role in the physiology of the mosquito. Apart from these results, the ATdel and ATattP mutants have the ability to produce eggs and undergo ovarian maturation. Further understanding of this neuropeptide can uncover its role in nutrition, development and its contribution to JH biosynthesis.

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