Fig. 3
From: Motoneurons innervation determines the distinct gene expressions in multinucleated myofibers
Motoneuron innervation is required for specialization of synaptic nucleus. A Whole-mount staining of TA muscles in ChATBCA-eGFP mice (2-month-old) after denervation for 3, 7, 16, or 30 days. BTX (red), DAPI (blue), ChAT-eGFP (green). Note that synaptic nucleus were gradually dispersed. D, denervation. B Quantification in A. n = 19 NMJs in Sham, n = 21 NMJs in D3, n = 21 NMJs in D7, n = 30 AChR cluster area in D16, n = 38 AChR cluster area in D30. Images were from three mice. One-way ANOVA with Tukey’s post hoc test for multiple comparisons F (4,127) = 26.46. C DAPI staining in SR/NSR in sham and denervated muscles. DAPI (blue), R-BTX (red). D Quantification in C. Sham, n = 63 nuclei in NSR and n = 63 nuclei in SR; D30, n = 74 nuclei in NSR and n = 49 nuclei in AChR cluster area. Images were from three mice. Two-way ANOVA with Tukey’s post hoc test for multiple comparisons. SR&NSR: F (1,245) = 183.3; Innervation & Denervation: F (1,245) = 22.01. E Whole-mount staining of TA muscles in 4-month-old SOD1G93A mice and littermate controls. BTX (red), DAPI (blue), SV2 (green). SV2 (Synaptic vesicle glycoprotein 2) is a synaptic protein to indicate presynapse at nerve terminals. F Quantification of synaptic nucleus number in SOD1G93A mice at indicated ages. Control mice: n = 25 NMJs (or images) in P60, n = 20 in P90, n = 21 in P120, n = 13 in P150. SOD1G93A mice: n = 25 NMJs (or images) in P60, n = 25 in P90, n = 26 in P120, n = 24 in P150. Images were from three littermate mice. Two-way ANOVA with Sidak’s post hoc test for multiple comparisons. Age: F (3,171) = 14.2; genotype: F (1,171) = 64.38. G Quantification of synaptic nucleus number in SOD1G93A mice at indicated ages. Control mice: n = 130 nuclei in NSR and 120 in SR in P60, n = 102 nuclei in NSR and 94 in SR in P90, n = 72 nuclei in NSR and 55 in SR in P120, n = 70 nuclei in NSR and 60 in SR in P150. SOD1G93A mice: n = 96 nuclei in NSR and 89 in SR in P60, n = 103 nuclei in NSR and 70 in SR in P90, n = 109 nuclei in NSR and 86 in SR in P120, n = 128 nuclei in NSR and 78 in SR in P150. Images were from three littermate mice. Two-way ANOVA with Tukey’s post hoc test for multiple comparisons. Age: F (3,1446) = 9.626; SR&NSR: F (3,1446) = 313.1. Unless otherwise specified, three independent experiments were performed; the mean ± SEM is shown. ***p < 0.001, ns, no significance