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Neuro2a cells co-transfected with FLAG-SOD1 and HA-CHGB were immunostained using anti-HA and anti-FLAG (C) or anti-misf SOD1 antibodies (B8H10) (D).

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Post-ANOVA Turkey test was used in each sex (same for all figures).

(B) The reduced sizes of α-motor neurons (α-MNs) in SOD13–4) was analyzed.

(B) The reduced sizes of α-motor neurons (α-MNs) in SOD1 mice (Fig. This effect was more pronounced in female than male mice (Fig. The neuromuscular junctions of gastrocnemius muscles were examined at 345 days of age.

It is noteworthy that female SOD1 mice are consistent with a precipitation of disease progression (Fig. As accumulation of misf SOD1 is known to induce ER stress in motor neurons of ALS (39,40), we examined the effects of variants CHGB variants caused an up-regulation of ER stress markers (Bip, p PERK, CHOP, cleaved caspase 12 (C12)) detected by Western blotting.

Most ALS patients die within 2 to 5 years after onset of symptoms since no effective treatment exists. The most frequent genetic causes of familial ALS are mutations in the gene coding for superoxide dismutase 1 (SOD1) (1,2) and an expansion of hexanucleotide repeats in noncoding region of are also known to be associated with ALS (5–26).

Chromogranins, which are major constituents of secretory large dense-core vesicles in neurons, act as chaperone-like proteins that bind mutant SOD1 proteins to promote their secretion (27).This finding led us to search for chromogranin genetic variations in ALS and control subjects.A few years ago we reported that a common chromogranin B (CHGB) variation was associated with higher risk to develop ALS and with earlier age of onset in both sporadic and familial ALS cases in cohorts of French-Canadian origin (28).These transgenic mice also exhibited similar excess CHGB protein levels in the microsomal fraction of the spinal cord when examined at 210 days of age ( showed no deficiency on the rotarod test or on body weight loss until 500 days of age (data not shown).The disease onset, as defined by 30% loss of maximum recorded time on rotarod (37,38), occurred at mean age of 365.7 ± 3.0 days in SOD1 mice (Fig. Furthermore, we have stratified the tested group of mice according to their sex status and determined whether sex had an effect of disease onset by analyzing rotarod performance. To investigate the degeneration of spinal α-motor neurons, lumbar cord sections at 345 days of age were immunostained with both anti-Ch AT and anti-Neu N antibody.Neuro2A cells were co-transfected with FLAG-tagged SOD1 and HA-tagged CHGB.


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