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. 2019 Dec 6;10:5575. doi: 10.1038/s41467-019-13165-7

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 10 — a–f Top: representative z-projections of boutons of the indicated genotypes labeled with HRP (magenta) and Gbb-HA (green). Genotypes specifically are: control (D42>gbb-HA), α₂δ-3^DD106/k (α₂δ-3^DD106/k10814; D42>gbb-HA), and α₂δ-3^DD196/k (α₂δ-3^DD196/k10814; D42>gbb-HA). Scale bar: 1 µm. Middle: individual Gbb-HA channel shown in grayscale. Membrane-proximal Gbb-HA was measured between the two dashed white lines. Bottom: individual Gbb-HA channel with inverted colors. g Quantification of intracellular Gbb-HA normalized to control levels before (2 mM KCl) and after (90 mM KCl) neuronal stimulation. For all Gbb-HA release experiments, n is the number of NMJs scored. h Quantification of membrane-proximal Gbb-HA (within 400 nm) normalized to control levels before and after neuronal stimulation. n.s. not significantly different. **p < 0.01; ***p < 0.001 by a two-tailed Mann–Whitney test. i Proposed model for α₂δ-3 acting as a physical barrier to promote Gbb signaling. Upon release from the neuron, Gbb is released into the synaptic cleft. With the aid of α₂δ-3, Gbb remains in close proximity to the presynaptic membrane and is then able to activate the BMP Type II receptor Wit. The receptor complex in turn phosphorylates the transcription factor Mad, transducing a BMP signal back to the nucleus of the neuron.