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. 2014 Mar 31;204(7):1141–1156. doi: 10.1083/jcb.201310090

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© 2014 Kasprowicz et al.

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Figure 7. — Invaginated pits in shi^ts1 mutants are converted into bulk cisternae upon Dynamin photoinactivation at the ultrastructural level. (A–D) Electron micrographs of shi^ts1/Y; shi-4C larvae treated with FlAsH for 10 min (+; B–D) or not treated (−; A) and illuminated for 2 min (+; C and D) or not illuminated (−; A and B) at permissive (RT; A) or restrictive (33°C) temperature (B–D) stimulated for 5 min with KCl. Bars: (A–C) 0.5 µm; (D) 0.25 µm. Arrowheads, submembrane inclusions; arrows, T bar; m, mitochondria; asterisks, invaginated pits. (E and F) Model of a bouton after surface rendering of a tomogram of shi^ts1/Y; shi-4C larvae after FALI (+/+) at restrictive temperature 33°C stimulated for 5 min with KCl (see also Video 1). (E) Note that some of the membrane inclusions are so massive that they are intertwined and folded inside each other. (F) Individual tomography models of different membrane inclusions. Gray, plasma membrane; blue, red, yellow, purple, and green, membrane inclusions. (G–I) Quantification of the number of synaptic vesicles <80 nm per area (G), the number of synaptic cisternae >80 nm per area (H), and the number of invaginated pits per area (I) in shi^ts1/Y; shi-4C larvae at 22°C (RT) not treated with FALI (−/−), at 33°C not treated with FALI (−/−), and at 33°C after FALI (+/+). Error bars show SEMs; ANOVA (post hoc Tukey’s test): *, P < 0.01; ***, P < 0.0001. n = 13, 10, and 14 bouton profiles from three larvae each.