investigating the molecular mechanisms that underlie tiling in drosophila r7 photoreceptors jennifer...

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Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

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Page 1: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Investigating the molecular mechanisms that underlie tiling in

Drosophila R7 photoreceptorsJennifer Salamé

George Fox University

Page 2: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Topographical maps: a major principle of organization in the nervous system

cell body

synapse

These synapses do not overlap (i.e. are "tiled").

Page 3: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Human retina is an example of a topographical map

Page 4: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Fly R7 photoreceptors: a great model to study tiling

Page 5: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Neuron circuit development occurs through two steps: axon pathfinding and synapse formation

Page 6: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Neuron circuit development occurs through two steps: axon pathfinding and synapse formation

Synapse Formation

1. filopodia search 2. cell adhesion stabilizes

3. pre-synaptic componentsaccumulate

4. post-synaptic assembly

Page 7: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

R7 axons require an Activin signal to tile

Wild-type R7s imp3 mutant R7s

Page 8: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Test the hypothesis that excessive synapse formation causes the tiling defect

Page 9: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Test the hypothesis that excessive synapse formation causes the tiling defect

Excessive synapse formation

Axon pathfinding error

Page 10: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Necessary

Approach 1: test whether synapse formation is necessaryfor the tiling defect

Page 11: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Necessary

Not necessary

Approach 1: test whether synapse formation is necessaryfor the tiling defect

Page 12: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Create syd-1, imp3 R7s

Wild type

imp3 mutant:tiling defect

imp3, syd-1double mutant:?????

syd-1 mutant:cannot form synapses

Page 13: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

imp3

syd-1

Test for non-complementation

Create a syd-1, imp3 double mutantchromosome

syd-1

Bal

xBal

imp3 Bal

Bal

x

imp3 syd-1

Bal

Page 14: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

syd-1, imp3 has a tiling defect

Page 15: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Approach 2: test whether excessive synapse formation is sufficient to cause a tiling defect

Page 16: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Approach 2: test whether excessive synapse formation is sufficient to cause a tiling defect

yes sufficient

not sufficient

Page 17: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

C. elegans rsy-1 mutant neurons form excessive synapses

Page 18: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Create drsy-1 mutant R7s in Drosophila

FRT82 drsy-1(Del)

Bal

x

xdrsy-1(Del)

FRT82 Bal

Bal

FRT82 drsy-1(Del)

Bal

Test with PCR and eye color

FRT82 Positive Control Candidates

Bal

Page 19: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

drsy-1 mutants do not have a tiling defect

Page 20: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Effect on synapse formation of drsy-1 mutation is inconclusive

Page 21: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Conclusions• Synapse formation is not necessary for a tiling defect• Excessive synapse formation may not be sufficient to

cause a tiling defect• Disrupting Activin pathway seems to affect axon

pathfinding

Axon pathfinding error

Page 22: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Future Research Directions

• Quantify synapses in drsy-1 mutants• Quantify synapses in tiling defect mutants• Identify genes regulated by Activin pathway in

R7s

Page 23: Investigating the molecular mechanisms that underlie tiling in Drosophila R7 photoreceptors Jennifer Salamé George Fox University

Acknowledgments

• UO Institute of Molecular Biology

• Dr. Tory Herman• Jennifer Jeffress• Erik Lyons• Jon Kniss• Scott Holbrook

•U

niversity of Oregon SPU

R Program

•D

r. Peter O’D

ay

•Chelsie Fish

•O

ther SPUR Interns!