Fig. 6: The absence of Cx43 and Cx45 in cultured skeletal myofibers increases the percentage of innervation in co-culture. DRG explants were co-cultured with skeletal myofibers.
From: Active acetylcholine receptors prevent the atrophy of skeletal muscles and favor reinnervation
a DRG explants were cultured in Campenot chambers in the central compartment (C), at 5 days of DRGs culture; skeletal myofibers are added in the lateral compartments (L) together with the distal axons (0 days of co-culture), up to 10 days of DRGs culture (5 days of co-culture). b Immunofluorescence of myofibers from Cx43fl/flCx45fl/fl mice under control conditions (Control cond.; top row), or treated with morpholinos to Cx43 and Cx45 (Cx43 + Cx45 Mor; middle row), and Cx43fl/flCx45fl/fl:MC mice (down row) at 5 days of co-culture of DRGs and myofibers, using antibodies against neurofilament 200 (NF; red; left column), and nicotinic acetylcholine receptor epsilon subunit (AChRε; green; middle left column). Merged images to identify innervated myofibers (middle right column). Arrows: innervated myofibers, arrowheads: non-innervated myofibers. Scale bar: 100 μm. Dotted rectangle is the magnification of innervated myofibers (right column). c Percentage of innervated myofibers. N = 5; at least 35 myofibers recorded in each independent experiment, each value is the mean ± SEM. The results obtained from Cx43fl/flCx45fl/fl mice are represented by white bars and from Cx43fl/flCx45fl/fl:MC mice by black bars. ***p < 0.001, for Cx43 + Cx45 Mor compared with Control cond. ###p < 0.001, for 43fl/flCx45fl/fl:MC compared with Control cond. by two-way ANOVA with Bonferroni post hoc test.
