Figure 4. M6 or MBγ activation induces a persistent calcium response in aSP13.

(A) Experimental protocol for M6 activation and aSP13 imaging, and representative calcium responses in aSP13 axons in the γ5 compartment. Scale bar, 10 μm. (B) Average ΔF/F responses in aSP13 axons, mean ± s.e.m. n = 22. (C) Average ΔF/F responses during the SFOCatCh ON periods in trials with (n = 9) or without 50 μM D-AP-5 (n = 22). ***p<0.001, t-test. (D) Average ΔF/F responses, imaged at 1 Hz after 200 s of post-stimulus section. (E) Experimental protocol for MBγ activation and aSP13 imaging, and representative calcium responses in aSP13 axons in the γ5 compartment. Scale bar, 10 μm. (F) Average ΔF/F responses in aSP13 axons, mean ± s.e.m. n = 13. (G) Average ΔF/F responses during the SFOCatCh ON periods in trials with or without 1 mM DA, 50 μM AP-5, or 150 μM mecamylamine (Mec). n = 13, 13, 5,10, respectively. **p<0.01, ***p<0.001, t-test. (H) Average ΔF/F responses, imaged at 1 Hz after 200 s of post-stimulus section. (I) Suppression indices (SI) of male flies in which shi^ts is expressed in M6 neurons, shifted to 32°C during training or testing, as indicated. ***p<0.001, **p<0.01, *p<0.05, permutation tests, see Supplementary file 2.

Figure 4—figure supplement 1. Stimulation of aSP13 does not elicit a persistent autonomous calcium response.

Average ΔF/F responses in the γ5 compartment of males expressing both SFOCatCh and GCaMP6s in aSP13 neurons. Experimental protocol as in Figure 3A. Mean ± s.e.m. n = 6.

Figure 4—figure supplement 2. Response in M6 after repetitive activation of MBγ is blocked by NMDA-R antagonist.

Average ΔF/F responses during the SFOCatCh ON periods with or without 50 μM AP-5. Mean ± s.e.m. n = 6 and 9, respectively.