Figure 5: Histopathological change in the CNS of Hsf-1-knockout AR-97Q mice.

(a–d) Histopathological analyses of AR-97Q and Hsf-1-knockout AR-97Q mice (13 weeks old). Immunohistochemistry for ChAT in the anterior horn of the spinal cord (a). Immunohistochemistry for calbindin in Purkinje cells (b). Immunohistochemistry for NeuN in the striatum (c). Nissl staining of the cerebral cortex (d). (e–h) Quantitative analysis of the size of neurons revealed that the neurons in each part of the CNS became atrophied in the Hsf-1-knockout AR-97Q mice compared with the AR-97Q mice. (i) Immunoblotting for ChAT in the spinal cord of AR-97Q and Hsf-1-knockout AR-97Q mice (13 weeks old). (j) Quantitative analysis of the signal intensity of the ChAT-immunoreactive bands. (k) Immunofluorescent staining of NMJs in 13-week-old AR-97Q and heterozygous Hsf-1-knockout AR-97Q mice (red, bungarotoxin; green, synaptophysin and phopsho-neurofilament H).The terminal of motor axons (green) are merged with virtually all the acetylcholine receptors labelled by bungarotoxin (red) in wild-type mice, indicating that NMJs are fully innervated. By contrast, some NMJs of AR-97Q mice lack synaptophysin and phopsho-neurofilament H staining owing to denervation (arrowheads), and this phenomenon was further enhanced by Hsf-1 depletion. (l) The frequency of denervation at NMJ is significantly increased in heterozygous Hsf-1-knockout AR-97Q mice. (m) Immunohistochemistry for Gfap in the anterior horn of the spinal cord. (n) Quantitative analysis of anti-Gfap immunoreactivity. (o) Immunoblotting for Gfap in the spinal cord of AR-97Q and Hsf-1-knockout AR-97Q mice (13 weeks old). (p) Quantitative analysis of the signal intensity of the Gfap-immunoreactive bands. Unpaired t-test. More than 500 neurons from three brains were analysed in each group (e–h), n=3 (j). Unpaired t-test. n=3 for each group (l), n=5 (n). Error bars indicate s.e.m. (e–h,j,l,n,p). Scale bars, 50 μm (a–d,k,m).