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Jiangnan Luo Lee Lab 30 Dec 2014 1 Lab meeting. Ongoing projects 2 1.Single cell morphology of lobular neurons Screening Gal4 lines (Janelia Farm & Kyoto.

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Presentation on theme: "Jiangnan Luo Lee Lab 30 Dec 2014 1 Lab meeting. Ongoing projects 2 1.Single cell morphology of lobular neurons Screening Gal4 lines (Janelia Farm & Kyoto."— Presentation transcript:

1 Jiangnan Luo Lee Lab 30 Dec 2014 1 Lab meeting

2 Ongoing projects 2 1.Single cell morphology of lobular neurons Screening Gal4 lines (Janelia Farm & Kyoto DGRC) Cell types and numbers labeled by Gal4 lines Dendritic morphology of single cell (lobula layer, dendritic field, orientation…) 2. Morphology of Tm5c is activity-dependent on photoreceptors? (R1-6/7-8) 3. Screening Mutants that affect dendrites of Tm20 neurons

3 3 Downstream targets for trans-medulla neurons (Tm5/20) Lin et al., 2014 (unpublished data) A P Li Ventral view Layer Dendritic field Orientation

4 4 Janelia Farm, FlyLight NC82 GFP D V ~ 12/lobe Li3 (R43E05) ~30 / lobe (Clustered) Li4 (R89D06) Anterior view

5 5 Li4 (NP1582) Otsuna et al., 2014 Frontiers in Neural Circuits

6 6 Li5 (R84C01) Janelia Farm, FlyLight Li5: ~20 / lobe NC82 GFP

7 Ongoing project 1 7 Single cell morphology of lobular neurons Li3/4/5-Gal4 > hsp1-FLP; UAS-FRT-CD2-y + -FRT-mCD8-GFP 3 rd instar larvae (mid- / late-) Adult (2-4 days old) Heat shock (2-3 min, 37°C) Dissect Li3

8 8 Single cell morphology – Li4 (R89D06) Mi T/Y? 10μm 24B10 Fas III GFP

9 9 Single cell morphology – (Li3/4/5) Li3 Con GFP 34 1 2 Li4: Lo4, 5 20μm Li5 Ventral view A P Li3: Lo5,6 Li4 (NP1582) Li5: Lo4, 5,6

10 10 Single cell morphology – (Li3/4/5) Li3Li4 Li5

11 11 Whole pattern – (Li3/4/5) Li3 Li5 1 2 3 4 Fas III Con GFP 20μm #1 #2 #3 #1 #2 N=12 N=11 N=13 N=19 N=21

12 12 Li5 (R84C01) Li3 (R43E05) Gal4 expression in the larval CNS Janelia Farm, FlyLight

13 13 Whole pattern – (Li3/4/5) Li4 (NP1582) GFP 24B10 FasIII Lin et al., 2014 (unpublished data) GFP 24B10 Pupae

14 Single cell morphology 14 Gal4 lines Sample Number Positive hitsPercentageCell type flipped out Heat shock time (min) Li3 (R43E05) 27415%Li2 40615%Li2.5 5958%Li3 Li4 (R89D06) 1700%Mi2 1200%Mi, T?2.5 Li5 (R84C01) 2827%Li2 7257%Li2.5 2115%Li3 Li4 (NP1582)301033%Li2

15 15 Future work – Project 1 1.Behavior study 2.Check Li3-Gal4 expression under developmental stages Manipulation of Li3 and test UV preference

16 Ongoing project 2 16 1. Single cell morphology of lobular neurons 2. Morphology of Tm neurons is activity-dependent? (on R1-6/7-8) De- or hyper- polarization of photoceceptors -> Dendrites of Tm5/20 neurons 3. Screening Mutants that affect dendrites of Tm neurons L-GMR-Gal4; NaChBac or L-GMR-Gal4; Kir2.1 > hsp1-FLP; C1-LexA; pLots2-stop-cd2-cherry, pLots2-stop-cd8-gfp

17 Morphology of Tm neurons are activity dependent on R7-8 ? 17 Increase sodium conductance Depolarization (NaChBac, bacterial sodium channel) Flies ready, soon have the results Hyperpolarization (Kir2.1, Inward-rectifying potassium channel) Prevent membrane polarization Genetics almost done L-GMR-Gal4; NaChBac> hsp1-FLP; C1-LexA; pLots2-s-cd2-cherry, pLots2-s-cd8-gfp L-GMR-Gal4; Kir2.1 > hsp1-FLP; C1-LexA; pLots2-s-cd2-cherry, pLots2-s-cd8-gfp

18 Ongoing project 3 18 1.Single cell morphology of lobular neurons 2. Morphology of Tm neurons is activity-dependent? (on R1-6/7-8) 3. Screening mutants that affect dendrites of Tm20 neurons

19 Ongoing project 3 19 Screening Mutants that affect dendrites of Tm20 neurons Mutant -FRT > hsp122-FLP; FRT40,42,G13-Gal80; ortC1-gal4, UAS-cd8-cherry Late 3 rd instar larvae Adult (2-4 days old) Heat shock (1 min, 37°C) Dissect

20 20 Screening Mutants that affect dendrites of Tm neurons Sema1a -/- Tm20 Tm5(a/b) 24B10 RFP

21 21 Mutants from Yuh Nung Jan’s Lab that affect dendritic development of PNS neurons

22 22 Future work – project 2 & 3 1.Dendritic study 2. Activity dependent study Hyper- or de-polarization of photoceceptors ---> Dendrites of Tm5c Request / search mutants (Jan lab & Kyoto stock center) Test the effects of mutations on Tm20 neurons


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