Presentation Title

Genetic Analysis of Two Potential Suppressors of tba-1(ju89)dlk-1(tm2024) Neural Defects

Faculty Mentor

Renee Baran

Start Date

17-11-2018 8:30 AM

End Date

17-11-2018 10:30 AM

Location

CREVELING 116

Session

POSTER 1

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

The microtubule cytoskeleton is crucial for the development and function of neurons including transport of synaptic components and vesicles. Regulation of microtubule (MT) dynamics in developing and mature neurons and how MTs form different types of arrays in neurons is not well understood. Our lab showed that a gain-of-function mutation in the alpha-tubulin gene tba-1(ju89) results in axon and synapse defects in both inhibitory GABAergic and excitatory cholinergic motor neurons in C. elegans (Baran et. al., 2010). Kurup et al. (2015) found that tba-1 in combination with dlk-1(tm2024) blocks synaptic remodeling of the DD motor neurons. In wild-type C. elegans, these neurons change their synapses from ventral muscles to dorsal muscles. The goal of this study is to determine if two potential suppressors of tba-1(ju89) also suppress the synaptic remodeling defects of the tba-1dlk-1 double mutant. Because the suppressors have no phenotype on their own, I am using balancer chromosomes marked with GFP in a series of genetic crosses to make C. elegans strains with the right genotypes and synaptic markers for the remodeling experiment. Suppressor/balancer strains were successfully created for each suppressor, and crosses to complete the tba-1(ju89)dlk-1(tm2024); suppressor triple mutants are in process. One of the suppressors has been cloned and codes for an E3 ubiquitin ligase involved in protein degradation. I designed PCR primers that will be used to amplify and sequence the region around the mutation in this gene to confirm that the suppressor mutation is still present in the triple mutant strain. Fluorescent microscopy will then be used to analyse the DD synapse remodeling pattern.

References

Baran, Renee, et. al (2010). PloS One 5: e9655.

Kurup Naina, et. al (2015). Curr Biol 25 (12): 1594-1605.

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Nov 17th, 8:30 AM Nov 17th, 10:30 AM

Genetic Analysis of Two Potential Suppressors of tba-1(ju89)dlk-1(tm2024) Neural Defects

CREVELING 116

The microtubule cytoskeleton is crucial for the development and function of neurons including transport of synaptic components and vesicles. Regulation of microtubule (MT) dynamics in developing and mature neurons and how MTs form different types of arrays in neurons is not well understood. Our lab showed that a gain-of-function mutation in the alpha-tubulin gene tba-1(ju89) results in axon and synapse defects in both inhibitory GABAergic and excitatory cholinergic motor neurons in C. elegans (Baran et. al., 2010). Kurup et al. (2015) found that tba-1 in combination with dlk-1(tm2024) blocks synaptic remodeling of the DD motor neurons. In wild-type C. elegans, these neurons change their synapses from ventral muscles to dorsal muscles. The goal of this study is to determine if two potential suppressors of tba-1(ju89) also suppress the synaptic remodeling defects of the tba-1dlk-1 double mutant. Because the suppressors have no phenotype on their own, I am using balancer chromosomes marked with GFP in a series of genetic crosses to make C. elegans strains with the right genotypes and synaptic markers for the remodeling experiment. Suppressor/balancer strains were successfully created for each suppressor, and crosses to complete the tba-1(ju89)dlk-1(tm2024); suppressor triple mutants are in process. One of the suppressors has been cloned and codes for an E3 ubiquitin ligase involved in protein degradation. I designed PCR primers that will be used to amplify and sequence the region around the mutation in this gene to confirm that the suppressor mutation is still present in the triple mutant strain. Fluorescent microscopy will then be used to analyse the DD synapse remodeling pattern.

References

Baran, Renee, et. al (2010). PloS One 5: e9655.

Kurup Naina, et. al (2015). Curr Biol 25 (12): 1594-1605.