FIG. 4.
Change in the distribution of mitochondria over time in living embryos with altered pHi. A) These single optical sections of live 2-cell embryos labeled with Mitotracker and imaged with TPLSM illustrate how mitochondria move away from their perinuclear localization shortly after the addition of the pHi-altering compounds, while embryos cultured in HECM-10 retain the perinuclear configuration. The HECM-10 embryo shown in the top row was cultured in the same dish as the HTMA-cultured embryo shown in the time series in the second row. The HECM-10 embryo shown in the bottom row was cultured in the same dish as the HDMO-cultured embryo shown in the time series in the third row. The t = 0-h images (left column) were collected prior to the addition of the pHi-altering compounds. Scale bar = 50 µm. The graph (B) illustrates the change of mitochondria distribution over time in living embryos. The bar graph shows data from individual images from each of the three treatments collected prior to the time-lapse imaging (0 h), as well as at 3 and 6 h following the addition of the pHi-altering compounds. The data shown represent six replicates. *Statistical difference (P < 0.05) from control within a time point. Bars indicate standard error. The line graph shows the quantitation of the mitochondrial pattern of individual images from a time-lapse sequence on a single culture. Each line represents three replicates. Bars indicate standard error. Arrows show the first time point that is statistically different (P < 0.01) from Time 0 within a treatment.