Accurate chromosome segregation at mitosis is ensured both by the intrinsic fidelity of the mitotic machinery and by the operation of checkpoints that monitor chromosomemicrotubule attachment. When unattached kinetochores are present, anaphase is delayed and the time available for chromosome-microtubule capture increases. Genes required for this delay first were identified in budding yeast (the MAD and BUB genes), but it is not yet known how the checkpoint senses unattached chromosomes or how it signals cell-cycle arrest. We report the isolation and analysis of a murine homologue of BUB3, a gene whose deletion abolishes mitotic checkpoint function in Saccharomyces cerevisiae. mBub3 belongs to a small gene family that has been highly conserved through evolution. By expressing recombinant proteins in insect cells, we show that mBub3, like yeast Bub3p, binds to Bub1 to form a complex with protein kinase activity. During prophase and prometaphase, preceding kinetochore-microtubule attachment, Bub3 localizes to kinetochores. High levels of mBub3 remain associated with lagging chromosomes but not with correctly aligned chromosomes during metaphase, consistent with a role for Bub3 in sensing microtubule attachment. Intriguingly, the number of lagging chromosomes with high Bub3 staining increases dramatically in cells treated with low (and pharmacologically relevant) concentrations of the chemotherapeutic taxol and the microtubule poison nocodazole.

Sperm cells from 45 infertile patients were investigated for disomy rates of chromosomes 1, 7, 10, 17, X and Y as well as for diploidy by single- and double-target in-situ hybridization. The patients who attended the infertility clinic were aged 23-46 years. Semenograms showed that the patients had oligo-, astheno-, oligoastheno-, oligoterato-, oligoasthenoterato-, or asthenoteratozoospermia. The average disomy rates determined in the patients were similar for all chromosomes, ranging from 0.10% (chromosome Y) to 0.14% (chromosomes 10 and X). Diploidy was detected with a mean incidence of 0.1%. With the exception of two patients who exhibited significantly increased diploidy rates of 0.35 and 1.6%, neither disomy nor diploidy was increased in the group of infertile patients as compared to healthy, fertile males.

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Sertoli cells of adult male laboratory mice were examined with a number of banding techniques and by nonradioactive in situ hybridization applying different repetitive DNA probes. All banding methods revealed the typical features of mouse Sertoli cells, i.e., a central nucleolus, usually with two chromocenters associated at diametrically opposed sides in which the centromeric regions of the chromosomes are clustered. Silver staining as well as in situ hydridization with rDNA labeled part of the chromocenters and the nucleolus, indicating transcriptional activity of at least some of the nucleolus organizer regions. In situ hybridization with X- and Y-specific DNA probes showed both sex chromosomes to be undercondensed in Sertoli cells This decondensation suggests expression of sex chromosomal genes in Sertoli cells. While the X chromosome appeared to occupy a central position near one of the chromocenters, the Y chromosome was found at the periphery of the nucleus in the majority of cells. Hybridization with telomeric sequences resulted in strong labeling of the chromocenters and dispersed signals at the nuclear periphery.

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The chromosomes of Brachidontes rodriguezi were analysed by means of direct Giemsa staining, silver staining, fluorescent in-situ hybridization (FISH) with 18S + 28S rDNA probes, replication banding and chromomycin A3 (CMA) and DAPI fluorescence banding techniques. The diploid chromosome number in this species is 32 and the karyotype is composed of two pairs of metacentric chromosomes, 2 pairs of telo/subtelocentric chromosomes and 12 pairs of subtelocentric chromosomes. 18S + 28S rDNA clusters were located on the short arms of the two pairs of telo/subtelocentric chromosomes. The replication band pattern induced in this species facilitates chromosome pairing and differentiation. The nucleolar organizing regions (NORs) replicate late in the S phase and were associated with bright CMA fluorescence and dull DAPI fluorescence, but not all the four NORs showed bright CMA fluorescence in a given cell; intra- and interindividual variability was found for this character.

Abstract. Mytilus galloprovincialis were collected from an intertidal population in N W Spain in 1988, and chromosomes from the gill tissue of 37 individuals were studied. The present paper describes the banding pattern of a M.galloprovincialis population which enabled us to identify all pairs of chromosomes. Banding was induced by means of a 2 x SSC (0.3 M sodium chloride :0.03 M sodium citrate)/Giemsa-staining technique, and a diagrammatic representation was constructed based on mean number of bands. The application of this banding technique may prove useful in future research related to the cytogenetics and cytotaxonomy of mussels and other molluscs.

The chromosomes of the mussel Mytilus galloprovincialis were analysed by means of chromomycin A3 (CMA), distamycin A/DAPI (DA/DAPI), DAPI/actinomycin D (DAPI/AMD) and chromomycin A3/distamycin A/DAPI (CDD) fluorescence banding techniques, C-banding, silver staining, N-banding and in situ hybridization with 18S+28S rDNA and telomere probes. 18S+28S rDNA clusters were located on the telomeres of two pairs of submeta/subtelocentric chromosomes. The nucleolar organizing regions (NORs) were associated with bright CMA fluorescence, dull DAPI fluorescence and C- and N-positive bands, but not all four NOR-associated heterochromatin bands showed bright CMA fluorescence in a given cell; intra- and interindividual variability was found in this character. Additional non-ribosomal C-bands did not show any differential fluorescent behaviour.