Figure 4

Tracking sperm organelles after fertilization. Crosses and microscopy were performed as described in Figure 2. Mitotracker-labeled mitochondria and GFP vesicle counts were taken from maximum projection images. Images were used to determine whether mitochondria or GFP-positive vesicles were clustered (within 2 microns of each other) or dispersed. The stacked bar graphs indicate the percentage of embryos at each stage that showed the phenotype indicated. A) Mitochondrial numbers decrease most dramatically between the pronuclear and mitotic phases. B) GFP::Ub vesicles (assumed to be MOs) were counted from the same images used for mitochondrial counts. MO numbers decrease with the same kinetics as mitochondria. The slight increase seen in later embryos may indicate vesicles that are not sperm-derived. C). A trend toward dispersion of sperm-derived mitochondria coincides with the decrease in numbers as shown in A. D) Ubiquitinated MO vesicles also become dispersed as embryos develop. E) The clustering of MOs and mitochondria together was assessed. Initially, the mitochondria and MOs cluster together. Following the first mitosis, these two organelles are no longer clustered together. For each trait, ten embryos of each stage were analyzed.