Severe disorganization of cytoskeletal architecture in GAN mice. (A) Electron microscopic examination of the axoplasm of GAN nerves revealed a diminution in microtubule content (arrows), an abnormal orientation and an increase in the diameter of neurofilaments (individual neurofilaments indicated by an arrowhead are magnified in the inserts). (B-D) The quantification of the cytoskeletal alteration in 48 week-old GAN mice was performed in sciatic nerves, L5-ventral and dorsal roots (n = 4 mice per genotype; 4 axons per mouse; 3 random pictures of distinct regions per axon; representing a total of 12 fields per mouse). (B) The mean number of microtubules per field was significantly lower in GAN compared to WT nerves (*, p < 0.05, Mann-Whitney test). (C) The alteration of neurofilament orientation was assessed by the measurement of the circularity of individual neurofilaments (circ = 1 and circ<1 representing a perfect circle and an elongated shape, respectively). The left panel displays average circularity scores, measuring the general orientation of the neurofilaments, for individual mice in the three tissues (the mean score per genotype is represented by a bar). The right panels show the standard deviations of the average circularity scores for the same analysis, a measure that is representative of the variations in the orientation of individual neurofilaments within each tissue section (*, p < 0.05, Mann-Whitney test). (D) Neurofilament diameter is significantly increased in GAN mice (10 individual neurofilaments per field; *, p < 0.05, Mann-Whitney test).