Glucose 6-phosphate dehydrogenase deficiency enhances germ cell apoptosis and causes defective embryogenesis in Caenorhabditis elegans

Yang, H-C; Chen, T-L; Wu, Y-H; Cheng, K-P; Lin, Y-H; Cheng, M-L; Ho, H-Y; Lo, S J; Chiu, D T-Y

Increased oxidative stress and DNA oxidative damage by G6PD knockdown in C. elegans. G6PD-depleted cells are highly susceptible to oxidative stress-induced cytotoxic effect with the concomitant generation of reactive oxygen species (ROS) and oxidative damage. 11, 12 In order to test whether G6PD knockdown increased oxidative stress in C. elegans, the level of oxidative stress and oxidative damage were determined ( Figure 3 ). By incubating first-day C. elegans adult with ROS-sensitive fluorescent dye 2 0 , 7 0 dichlorodihydrofluoroscein diacetate (DCFDA), increased ROS was detected in Gi C. elegans compared with mock C. elegans (1.2-fold, P ¼ 0.038, N ¼ 6) (Figure 3a ). To further investigate whether G6PD deficiency caused oxidative damage in macromolecues, malondialdehyde (MDA) and 8-hydroxy-2-deoxyguanosine (8-OHdG) were measured. While there was no significant difference of MDA level between mock and Gi C. elegans (data not shown), 8-OHdG level was significantly increased in Gi C. elegans compared to mock (1.5-fold, P ¼ 0.005, N ¼ 6) (Figure 3b ). This finding is consistent with a previous report in G6PD-deficient cell model, 11 indicating that G6PD may play a role in protecting cells against DNA oxidative damage in C. elegans.