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DCN – their axonal behaviour

This overview presents research on axon extension and retraction, experiments on dorsal cluster neurons, axon positioning, computational models, and the impact of mutations like Chimo, Rab5.DN, Rab7.DN, dORK, and Shi.DN. Detailed data includes neuron and axon analysis, experimental conditions, genotype variations, and statistical results. The study also explores the computational model's algorithm, gene interactions, and hypotheses regarding axon distribution in Lobula and Medulla.

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DCN – their axonal behaviour

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  1. DCN – their axonal behaviour LabRetreat 04.06.2007

  2. Overview • Axon Extension & Retraction • Experiments: • Number of Dorsal Cluster Neurons • Axons in ME • Neuron vs. Axon positioning • Computation: • Wildtype/Mutation Overview • Chinmo • Axon Branching • Experiments • Rab5 DN • Rab7 DN • dORK • Shi DN

  3. Number of Dorsal Cluster Neurons • Genotype: w1118/yw;UAS-lacZ.NZ(20b);atoGal-14a & w1118/yw;;UAS-lacZ.NZ(J312)/atoGal-14a • Condition: room temperature • Sorting: • Age: 2, 7, 14, 28, 42 days • Gender: ♀ and ♂ • Staining: β-gal

  4. Number of Dorsal Cluster Neurons Genotype: atoGal4-14a X UAS::nlacZ Staining:α-βgal Gender: male Age: 2 days old Location: right side Neurons: 56

  5. Number of Dorsal Cluster Neurons → There is no statistical significant difference between the two Dorsal Cluster of each brain regarding age and gender.

  6. Graphical Results DCNs Mean + SD ** indicates statistical significant differences, (not all stat. signif. differences are shown)

  7. Axons in the Medulla • Genotype: w/+;;UAS-lacZ,atoGal-14a/+ • Condition: room temperature • Sorting: • Age: 2 days • Gender: ♀ and ♂ • Staining: β-gal

  8. Axons in the Medulla Genotype: atoGal4-14a X UAS::lacZ Staining:α-βgal 1 2 3 4 5 6 7 8 9 Gender: male Age: 2 days old Location: right side Axons: 12 10 11 12

  9. Distribution: Axons in Medulla

  10. I II III IV V VI VII VIII IX X XI XII Axons in Medulla

  11. Axons in Medulla

  12. Axons in Medulla

  13. Neuron vs. Axon Position • Genotype: w/yw,hsFlp;+/(Sp v Cyo); UAS-lacZ,ato-Gal4-14a/UAS>CD2,y>mCD8-Gfp • Condition: 28°C • Age: 2 days • Staining: β-gal (mouse), gfp (rabbit)

  14. a1 a2 a3 a4 a5 a1 a2 a3 a4 a5

  15. a1 a2 a3 a4 a5

  16. Computational Model • Assumptions • Number of Neurons: 56 • Number of Axons in Medulla: 12 • Algorithm • GIN • Neighbourhood Competition + Timing

  17. Wnt FGF Fz FGFR DSH RAC P-JNK P-JNK EXTENSIONRETRACTION Lobula Medulla Gene Interaction Network Srahna et al. 2006

  18. Computational Model - Overview Number Of DCNs Wnt based on Connectivity Of DCNs FGF GIN uses

  19. GIN – Implemented Logic before Lobula Lobula Medulla Time wnt5 & wnt2 & fz & fz2 Signals: None fgf & fgfr true false false true false Rac: false true true false true JNK:

  20. Neighbourhood Competition • Neighbourhood consists of • 8 + • the current neuron

  21. Neighbourhood Competition • Logic for competition • Choose one neuron ni randomly (it must have permission from GIN) • If (ni-4+ni-3+ni-2+ni-1+ ni+ni+1+ni+2+ni+3+ni+4 >=2) • Then ni→ Lobula = 0 • Else ni→ Medulla = 1 • Repeat these two steps until there is a stable pattern or a time dependent pattern has been found.

  22. wnt fgf Computational Model Lobula Medulla 1 2 3 4 5 6 7 8 9 10 11 12

  23. Computational Model

  24. Computational Model – Wnt Signal Mean + SD

  25. Computational Model – FGF Signal Mean + SD

  26. Chinmo Hypothesis: Only those axons are in the Medulla which lacks Chinmo !

  27. Chinmo • Assumptions: • 56 Neurons • 12 Axons remain in the Medulla • There is a neighbourhood communication • GIN extended by Chinmo

  28. Chinmo – A teamplayer of GIN? Wnt FGF Fz FGFR DSH Chinmo RAC P-JNK P-JNK EXTENSIONRETRACTION Srahna et al. 2006 Lobula Medulla

  29. GIN + Chinmo – Implemented Logic before Lobula Lobula Medulla Time wnt5 & wnt2 & fz & fz2 Signals: None fgf & fgfr & Chinmo true false false true false Rac: false true true false true JNK:

  30. Chinmo • Some thoughts: 56! 56! = 5,583833073 * 1011= 558.383.307.300 = 12! * 44! 12! * (56-12)! • Model: • 40.000 different settings (<< 1% ) • 1.019 settings had 12 Axons in the Medulla • 1.019 are not unique ! • 24 fits the spatial structure of „12 Axons in ME“ • BUT: Since these 24 settings are not unique, it might be that there is no competition and therefore CHINMO+GIN decide about axonal fate.

  31. Chinmo

  32. Comparision

  33. Axon Branching

  34. A18♀ X Rab5DN♂w;;UAS-lacZ,atoGal4-14a/UAS-Drab5S43N Rab5DN ♂ Wildtype

  35. Clathrin♀ X A18♂w/w1118chc4;;UAS-lacZ,atoGal4-14a ♀ FM7aGFP;;UAS-lacZ,atoGal4-14a ♂ Wildtype Clathrin ♀ Clathrin ♂

  36. A18♀ X Rab7DN♂w;UAS-Drab7;UAS-lacZ,atoGal4-14a Rab7DN ♀ Rab7DN ♂ Wildtype

  37. A18♀ X dORK♂w;;UAS-lacZ,atoGal4-14a/UAS dORKΔc1 dORK ♀ dORK ♂ Wildtype

  38. A18♀ X ShiDN♂w/yw,UAS-ShiK44a(4-1);UAS-ShiK44a(3-7);UAS-lacZ,atoGal4-14a ♀w;UAS-ShiK44a(3-7);UAS-lacZ,atoGal4-14a ♂ ShiDN ♀ ShiDN ♂ Wildtype

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