Combined immunocytogenetic and molecular cytogenetic analysis of meiosis I human spermatocytes

Abstract

We have used a combination of immunocytogenetic and molecular cytogenetic technology on human spermatocytes to investigate (1) meiosis I chromosome pairing, and (2) organization of synaptonemal complex (SC)-associated chromatin with respect to whole chromosome paints, unique DNA sequences and repetitive DNA of heterochromatic blocks, centromeres and telomeres. It is evident that synapsis normally starts at the termini of homologues. In general, synapsis proceeds synchronously from termini towards the centre of bivalents without any indication of interstitial initiation. Some aberrant meiosis I spermatocytes showed asynchronous pairing, demonstrating not only large differences in the degree of SC formation between bivalents, but also chromosome alignment without synapsis as well as clear interstitial synaptic initiation. It may be the case that alignment normally takes place along the entire length of homologues before synapsis occurs and that the potential for synaptic initiation exists along the length of chromosomes. Telomeric sequences were seen tightly associated with the SCs, as might be expected considering their kinetic properties in relation to the nuclear membrane. Other repetitive DNA, i.e. centromeric alpha-satellites and classical satellites of the heterochromatic blocks 1qh and 9qh, were all found to form loops that are associated with SCs only at their bases. A unique DNA cosmid probe (21q22.3) was found to produce a hybridization pattern consisting of spots located outside SC. The fluorescence in situ hybridization (FISH) signals of these spread DNA sequences have a granular appearance, probably reflecting the pattern of coiling and chromatin condensation of the target DNAs.