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. 2022 Apr 27;59(7):4044–4064. doi: 10.1007/s12035-022-02818-2

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© The Author(s) 2022

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visithttp://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — In (a) we show the percentage of singly- and polyinnervated NMJ after 4 applications over the LAL surface (one application every day between P5–P8 (observation at P9) of one of the following VGCC inhibitor substances: nitrendipine (NT 1 μM, an L-type channel blocker), ω-conotoxin-GVIA (ω-CON 1 μM, N-type channel blocker), and ω-agatoxin-IVA (ω-AGA 100 nM, P/Q-type blocker). Also, the L activator Bay-K8644 (5 μM), the P/Q- and N-type activator GV-58 (20 μM), and the intracellular calcium chelator BAPTA-AM (5 μM). The histogram in (b) shows the percentage of S1-S4 clusters in the untreated control mice (PBS) and after the 4 applications of the aforesaid substances. Data were presented as percentages of NMJ ± SD. Fisher’s test: * p < 0.05, ** p < 0.01, *** p < 0.005. The confocal images in (c) show examples of representative NMJ areas with singly, dually, and innervated by three or more axons (the corresponding number of asterisks) from YFP muscles. At the left, the L-type channel blocker nitrendipine (NT) delays axon loss because many multi-innervated NMJs persist. By the contrary, at the right, the L activator Bay-K8644 increases the number of monoinnervated junctions. The bar indicates 10 μm