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. 2024 Jun 7;15:4867. doi: 10.1038/s41467-024-48925-7

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© The Author(s) 2024

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 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, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — Quantifications of inhibitory synapses on spared motor neurons at P112 upon AAV8-hSYN-DIO-hESYT1 injections. Synaptic densities are normalized by motor neuron perimeter. Examples of Vgat positive synaptic densities in the different conditions are shown in (a–d). a, b Show synaptic density in WT and En1^cre mice respectively. c and (d) show differences in synaptic densities between SOD1^G93A and SOD1^G93A;En1^cre mice. e Synaptic density in SOD1^G93A;En1^cre is significantly increased compared to SOD1^G93A and comparable to the synaptic densities of WT and En1^cre mice (One-way ANOVA and Dunnett’s post hoc WT P = 0.045, En1^cre P = 0.0119, SOD1^G93A;En1^cre P = 0.0306, WT = 4, SOD1^G93A N = 5, En1^cre N = 4, SOD1^G93A;En1^cre N = 5). Merge microphotographs in (f–i) show examples of quantified Chat positive motor neurons in the different genotypes. Vgat in orange, Chat in magenta, DAPI in blue. Motor neuron quantifications performed at P112 in (j) WT, (k) En1^cre, (l) SOD1^G93A and (m) SOD1^G93A;En1^cre mice. Fluoro-Nissl in red. n SOD1^G93A;En1^cre mice show increased motor neuron survival upon ESYT1 overexpression when compared to the SOD1^G93A mice (One-way ANOVA and Dunnett’s post hoc SOD1^G93A; En1^cre P = 0.0080, WT P = 0.0023, N = 6). A minimum of 170 motor neurons per condition was quantified. En1^cre mice overexpressing hEsyt1 show a trend in lower number of large motor neurons in the lumbar spinal cord (One-way ANOVA and Dunnett’s post hoc, En1^cre P = 0.2732, N = 5). o Comparison of motor neuron size in En1^cre non-injected and En1^cre injected mice with AAV8-hSYN-DIO-hESYT1 shows shrinkage of motor neurons upon ESYT1 overexpression (t-test, two-tailed, P = 0.0073, En1^cre non-injected N = 4, En1^cre injected N = 5). Scale bar in a–d = 10 μm, f–i = 10 μm and in j–m = 50 μm. All graphs show mean values ± SEM. Source data are provided as a Source Data file.