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Figure 1 N-Glycosylation occupancy of Kv3.1b alters its spatial arrangement in motor neurons. (A) Schematic of a spinal hemi-segment illustrating the morphological innervation of the caudal primary motor neuron (CaP) of the ventral musculature. (B) A confocal projection (~40 µm stack) of one spinal hemi-segment illustrating the IHC co-staining of the Kv3.1b channel (Alexa Fluor 488, green) and Znp-1 (Alexa Fluor 555, red) in a CaP motor neuron. Arrowheads denote over-lapping expression of Kv3.1b and Znp-1. (C) Confocal projections of CaP neurons expressing EGFP, WT and N220Q Kv3.1b proteins. The bottom row shows confocal images, and top row images are overlaid with transmitted images to show the horizontal myoseptum (dashed line). (D) 3D digital rendering of the reconstructed neurons of images in (C) shown in both sagittal and transverse orientation. (E) Average axonal length, neuron axonal volume and number of nodes of CaP neurons expressing EGFP (n = 12), WT (n = 10) and N220Q (n = 11) Kv3.1b proteins. Graphs denote mean ± SEM and were compared by one-way ANOVA followed by Kruskal-Wallis adjustment (** p < 0.0005, *** p < 0.0001). See also Movies Figures S1–S3.