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. 2015 Jun 26;4(8):947–953. doi: 10.1242/bio.011262

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© 2015. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 3. — Arl1, Garz, and Arfip function in the same genetic pathway during synapse growth. (A-D) Immunofluorescent images of the neuromuscular junctions (NMJs) formed on muscle 4 from larvae of the indicated genotypes co-stained for presynaptic VGluT (green; insets i) and postsynaptic Discs-large (Dlg) (red; insets ii). Insets are higher magnification images of the terminal boutons of the synapse shown. Scale bar=10 μm. (E-I) Graphs represent the average number of boutons at the muscle 4 NMJs normalized to control values. (E) The expression of wild type Arfip (wt) but not a GTPase-binding mutant Arfip (GBD) rescued synapse growth defects in arfip^12/71 mutant neurons using either the Ok6-Gal4 (Ok6 rescue) or C155-Gal4 (C155 rescue). (F) Transheterozygotic genotypes consisting of Arl1¹ and arfip¹², Arl1^FRT and arfip¹² have reduced synapse growth compared to heterozygotic Arl1 controls (Arl1¹/+ and Arl1^FRT/+). Expression of the pUAS-Arl1-HA transgene in the Arl1^FRT/arfip¹² heterozygotic background using the OK6-Gal4 driver [Arl1^FRT/arfip¹² (+Arl1)] restores synapse growth to control levels compared to the control background harboring only the OK6-Gal4 driver [Arl1^FRT/arfip¹² (+ctrl)]. (G) Transheterozygotic genotypes consisting of Arl1^FRT and garz^W982 or of Arl1^FRT and garz¹³⁷ have reduced synapse growth compared to heterozygotic garz controls (garz^W982/+ and garz¹³⁷/+). (H) Transheterozygotic genotypes containing heterozygotic mutations in arfip (arfip¹² or arfip⁷¹) and garz¹³⁷ have reduced synapse growth compared to arfip/+. (I) The expression of a UAS construct encoding a dsRNA to garz (UAS-garz^RNAi) in motor neurons using either the Ok6-Gal4 motor neuron driver or the C155-Gal4 panneuronal driver results in reduced synapse growth. (J) Transheterozygotic genotypes between mutations in arfip⁷¹ and the exocyst genes sec5^E10 or sec15¹ have no effect on synapse growth. Error bars=s.e.m. *P<0.05; significant difference from controls by ANOVA.