Fig. 5

The impact of folate deficiency to embryonic cell proliferation and one-carbon pools.

(A-I) Embryos at the indicated stages were subjected to whole mount immunological staining with anti-pH3 antibodies for proliferating cells in whole body and especially in the cardiac area (dashed line). The green fluorescent puncta represent the positive signal of pH3, which were distinctive and distinguishable from the homogeneous green fluorescent background raised from the heat-shock induced EGFP-γGH. Images were taken from dorsal view (A-C) and lateral view (A’-C’ and D-I) with anterior to the left. (J) Quantification of phenotypes (normal, mild, and severe) for embryonic cell proliferation in each group was based on the signal intensity detected at 26 hpf, as shown in (A-C). Data were collected from at least 3 independent experiments with the total sample number of 9–15 for each group. Larvae of 31 hpf (K-M) and 3 dpf (N-P) were subjected to intracellular folate content measurement with HPLC. (Q) Larvae of 32 hpf (left) and 55 hpf (right) were analyzed for homocysteine content. Data presented were the average of at least three independent trials and were normalized by embryo weights. CTL, heat-shocked wild-type control; MFD, mild folate deficiency; SFD, severe folate deficiency; FD, folate deficiency. 5-CHO, 5-formyltetrahydrofolate; THF, tetrahydrofolate; 10-CHO-THF, 10-formyltetrahydrofolate; 5-CH3-THF, 5-methyltetrahydrofolate. *, p<0.05; **, p<0.01; ***, p<0.001.