Daniel Johansen Professor Tory Herman Mentor: Jen Jeffress UO Dept. of Biology and Institute of Molecular Biology Investigating the Role of Syd-1 in Synapse.

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Presentation transcript:

Daniel Johansen Professor Tory Herman Mentor: Jen Jeffress UO Dept. of Biology and Institute of Molecular Biology Investigating the Role of Syd-1 in Synapse Development

Neuronal Communication Requires Proper Synapse Development

We are Studying Synapse Development Using Fruit Flies

Wild-type Synapses

The Protein Syd-1 is Required for Normal Synapse Development Syd-1 Mutant SynapsesWild-type Synapses Sigrist et. al 2010

Syd-1 has Sequence Similarity to GAPs Inactive Active Gap proteins “turn off” GTPases GTPase CYTOSKELETON

Does Syd-1 have GAP activity? Inactive Active GTPase CYTOSKELETON Over-express GTPase + Defect Worsens Over-express GEF Over-express GTPase Defect in flies Over-express GTPase + Defect Improves Over-express GAP

We Used Eye Development as a Read- out of GTPase Activity Wild-type Eye Rough Eye Death Rougher Eye

There are Six Different RhoGTPases that Syd-1 Might Regulate Multiple Rho-type GTPases in Drosophila Rac1 Rac2 Mtl Rho1 RhoL Cdc42

Genetic Crosses Were Done to Test the Interaction of Syd-1 with Some RhoGTPases Over-express Rac1 Over-express Rho1 + Syd-1 ??? Over-express Cdc42 OE GTPase and Syd-1

Syd-1 Appears to Not be Regulating Cdc42 Cdc42Cdc42 and Syd1

Syd-1 May be Acting Like a GAP for Rac1 and Rho1 Rho1 Rho1 and Syd1 Rac1 Rac1 and Syd1

Syd-1 Acts Upstream of Liprin- α Syd-1 Liprin-α Scaffolding Liprin-α Function Syd-1 Localizes Liprin-α to Synapses Wild-typeSyd-1 Mutant Sigrist et. al 2010

Liprin- α Regulates Axon Transport

Without Liprin-α, Axon Transport Does Not Occur Normally P value < 0.001

Does Syd-1 Help Liprin-α Promote Axon Transport? Syd-1 Liprin-α Scaffolding Axon Transport Liprin-α Functions Liprin-α Syd-1 ?

Our Preliminary Data Suggested Syd-1 May Also be Required for Axon Transport P value =

Vesicle Tracking was Done to Determine Syd-1’s Role in Axon Transport

Results Showed Differences Compared to Wild-type Samples Percent Anterograde Movement of Syd-1 Percent Anterograde Movement of Wild-type

Bimodal Results Due to Location of Sampling Along the Axon Syd-1 Wild-type

Syd-1 Does Not Regulate Liprin-α in Axon Transport Syd-1 Liprin-α Scaffolding Axon Transport Liprin-α Functions Liprin-α

Future Study Confirm the GAP activity of Syd-1 with the GTPases in vitro Mutated Syd-1 that affects its GAP domain Is the GAP domain necessary for synapse development? Why is there a difference in movement speed between the location of sampling in the axon?

Acknowledgements Herman Lab Tory Herman Jennifer Jeffress Sasha Feoktistov Eric Lyons Jon Kniss Alex Whitebirch