Research Paper Volume 11, Issue 22 pp 10338—10355

Figure 4. Mitochondrial content under basal conditions in control, wt/ _N370SGBA and _N370SGBA/_N370SGBA neurospheres was assessed by western blotting for the mitochondrial proteins VDAC1 (outer mitochondrial membrane), SDHA (inner mitochondrial membrane), and TFAM (matrix), normalized by β-actin content. All markers tended to increase in _N370SGBA/_N370SGBA neurospheres in basal conditions, when compared to control lines (A) (SDHA was significant). As expected in control lines, protein levels of VDAC1, SDHA and TFAM and ATP levels decreased following CCCP treatment for 24 hours (B and C). Despite an apparent increase in mitochondrial mass in untreated conditions, ATP levels remained unchanged in wt/_N370SGBA and _N370SGBA/_N370SGBA neurospheres (C). Mitochondrial markers of wt/_N370SGBA did not show a marked response to CCCP uncoupling, in fact, TFAM significantly increased in the heterozygous lines and the decrease in ATP levels were not so pronounced (B and C) and ATP levels of homozygous _N370SGBA/_N370SGBA neurospheres showed a null response to CCCP uncoupling. All of the above data suggest that mitochondrial function may be impaired in GBA mutant neurospheres. AU: arbitrary units. Thin line indicates p value <0.05. Results are expressed by mean± SEM.