Presentation Title

Analyzing the Significance of DIP α and Dpr 10 at Wiring the Nervous System

Faculty Mentor

Kai Zinn and Shuwa Xu

Start Date

23-11-2019 10:00 AM

End Date

23-11-2019 10:45 AM

Location

87

Session

poster 3

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

Analyzing the Significance of DIP α and Dpr 10 at Wiring the Nervous System

By Joshua Morales and Mentors: Kai Zinn, PhD, and Shuwa Xu, PhD

In developmental neurobiology, neurons must make correct synaptic connections with other neurons for the nervous system to develop properly. Many studies have shown that cell-surface proteins are responsible for guiding neurons to their correct synaptic formation. The Zinn Lab at Caltech has previously identified two classes of interacting cell surface proteins, known as the defective proboscis extension response proteins (Dprs) and Dpr-interacting proteins (DIPs), that function in neuronal targeting and synaptic specificity in the visual system of the central nervous system (CNS) and neuromuscular system of Drosophila melanogaster. Here, we sought to examine the roles of DIP/Dpr protein interactions in neuronal development. This was done by modifying the genome sequence of D. melanogaster using CRISPR/Cas9 technology which weakened the binding affinity DIP alpha. Through various screening tests by gel electrophoresis and sequencing, we were able to ensure that the guide RNA, its donor plasmid, and the RMCE donor plasmid were correctly made for the CRISPR/Cas 9 edits. Embryo fluorescence staining was also utilized to visualize where Dpr proteins were being expressed. Thus far, we have selected the EGFP UAS line because it gave the strongest visual signal. Future work includes conducting crosses with this UAS line along with other Dpr lines to see where these Dprs are being expressed. Overall, this work will lead to a better understanding of neuronal synaptic specificity.

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Nov 23rd, 10:00 AM Nov 23rd, 10:45 AM

Analyzing the Significance of DIP α and Dpr 10 at Wiring the Nervous System

87

Analyzing the Significance of DIP α and Dpr 10 at Wiring the Nervous System

By Joshua Morales and Mentors: Kai Zinn, PhD, and Shuwa Xu, PhD

In developmental neurobiology, neurons must make correct synaptic connections with other neurons for the nervous system to develop properly. Many studies have shown that cell-surface proteins are responsible for guiding neurons to their correct synaptic formation. The Zinn Lab at Caltech has previously identified two classes of interacting cell surface proteins, known as the defective proboscis extension response proteins (Dprs) and Dpr-interacting proteins (DIPs), that function in neuronal targeting and synaptic specificity in the visual system of the central nervous system (CNS) and neuromuscular system of Drosophila melanogaster. Here, we sought to examine the roles of DIP/Dpr protein interactions in neuronal development. This was done by modifying the genome sequence of D. melanogaster using CRISPR/Cas9 technology which weakened the binding affinity DIP alpha. Through various screening tests by gel electrophoresis and sequencing, we were able to ensure that the guide RNA, its donor plasmid, and the RMCE donor plasmid were correctly made for the CRISPR/Cas 9 edits. Embryo fluorescence staining was also utilized to visualize where Dpr proteins were being expressed. Thus far, we have selected the EGFP UAS line because it gave the strongest visual signal. Future work includes conducting crosses with this UAS line along with other Dpr lines to see where these Dprs are being expressed. Overall, this work will lead to a better understanding of neuronal synaptic specificity.