After fusion with an egg, the haploid genome of a mammalian sperm expands from a volume of a few cubic microns to a volume of several thousand cubic microns in a fully developed zygote pronucleus. Using fluorescent in situ hybridization we studied two aspects of the chromatin organization of the sperm-derived genome during this process in a model system involving fusion of human sperm with hamster eggs. We found that (a) from the beginning of sperm head decondensation to early pronuclear stages, the hybridization signal of a probe targeted to the satellite III heterochromatin of chromosome 1 appeared as an irregularly shaped domain. In fully developed pronuclei, the signal became a fiber as much as 20-fold more extended compared with interphase somatic nuclei. The signal appeared as a string of non-uniformly distributed "beads" with interspersed gaps. A similar spatial distribution and appearance recurred after the first cleavage division. (b) Male- and female-derived genomes assembled non-randomly on the first mitotic spindle, and continued to be sequestered from each other in the nuclei of two-cell stages.