Leukocyte Nucleolus and Anisakis pegreffii—When Falling Apart Means Falling in Place

The view of the nucleolus as a mere ribosomal factory has been recently expanded, highlighting its essential role in immune and stress-related signalling and orchestrating. It has been shown that the nucleolus structure, formed around nucleolus organiser regions (NORs) and attributed Cajal bodies, is prone to disassembly and reassembly correlated to various physiological and pathological stimuli. To evaluate the effect of parasite stimulus on the structure of the leukocyte nucleolus, we exposed rat peripheral blood mononuclear cells (PBMC) to the crude extract of the nematode A. pegreffii (Anisakidae), and compared the observed changes to the effect of control (RPMI-1640 media), immunosuppressive (MPA) and immunostimulant treatment (bacterial lipopolysaccharide (LPS) and viral analogue polyinosinic:polycytidylic acid (poly I:C)) by confocal microscopy. Poly I:C triggered the most accentuated changes such as nucleolar fragmentation and structural unravelling, LPS induced nucleolus thickening reminiscent of cell activation, while MPA induced disassembly of dense fibrillar and granular components. A. pegreffii crude extract triggered nucleolar segregation, expectedly more enhanced in treatment with a higher dose. This is the first evidence that leukocyte nucleoli already undergo structural changes 12 h post-parasitic stimuli, although these are likely to subside after successful cell activation.

Meiotic Recombination in the Giraffe (G. reticulata)

Recently, the reticulated giraffe (G. reticulata) was identified as a distinct species, which emphasized the need for intensive research in this interesting animal. To shed light on the meiotic process as a source of biodiversity, we analysed the frequency and distribution of meiotic recombination in 2 reticulated giraffe males. We used immunofluorescence detection of synaptonemal complex protein (SYCP3), meiotic double strand breaks (DSB, marked as RAD51 foci) in leptonema, and crossovers (COs, as MLH1 foci) in pachynema. The mean number of autosomal MLH1 foci per cell (27), which resulted from a single, distally located MLH1 focus observed on most chromosome arms, is one of the lowest among mammalian species analysed so far. The CO/DSB conversion ratio was 0.32. The pseudoautosomal region was localised in the Xq and Yp termini by FISH and showed an MLH1 focus in 83% of the pachytene cells. Chromatin structures corresponding to the nucleolus organiser regions were observed in the pachytene spermatocytes. The results are discussed in the context of known data on meiosis in Cetartiodactyla, depicting that the variation in CO frequency among species of this taxonomic group is mostly associated with their diploid chromosome number.

The Dicentric Chromosome dic(20;22) Is a Recurrent Abnormality in Myelodysplastic Syndromes and Is a Product of Telomere Fusion

We describe a recurrent dicentric chromosome formed by telomere fusion between chromosome 20 and chromosome 22 in 4 cases of myelodysplastic syndromes (MDS) or acute myeloid leukaemia (AML). In particular, the presence of residual telomere sequences at the site of translocation in 3 of the 4 cases makes a compelling case for telomere fusion. This is the first description of a recurrent telomere fusion event in any malignant condition. The 20q subtelomeric region was retained in all 4 examples despite deletion of the 20q12 region closer to the centromere. The original dicentric chromosome in all 4 cases contained nucleolus organiser region material from the short arm of chromosome 22 and had also undergone secondary rearrangements that produced amplification of the common gained region on 20q. We propose that the sequence of events producing this chromosome abnormality is: degradation of the telomeres, formation of an unstable dicentric chromosome by 20q and 22p telomere fusion, breakage-fusion-bridge cycles causing copy number aberration between the centromeres, selection of cells with 20q12 deletion, and further selection of cells with 20q11.2 gain. The last 2 steps are driver events responsible for the abnormal chromosomes found in the malignant cells. Finding recurrent patterns in the complex genome reorganisation events that characterise poor-prognosis, complex-karyotype AML and MDS will help us understand the mechanisms and oncogenic driver mutations in these poorly understood malignancies.

Karyotype of Araucaria angustifolia and the decondensation/activation mode of its nucleolus organiser region

The chromosomes of the gymnosperm Araucaria angustifolia (Bertol.) Kuntze were analysed with the fluorochromes chromomycin A3 (CMA) and 4′,6-diamidino-2-phenylindole (DAPI) and with C-banding. This species contains a diploid complement made of 26 chromosomes, with 18 larger metacentric, four smaller metacentric and four submetacentric chromosomes. The only CMA+/DAPI– region observed corresponded to the nucleolus organiser region (NOR) localised at the proximal portion of a large metacentric chromosome pair. C-banding marked the NOR as well as a terminal region of another chromosome pair. In addition, small C-bands were occasionally seen interspersed in many chromosomes. The NOR appeared to condense at approximately the same rate as the rest of the chromosome from prophase throughout metaphase. In interphase nuclei, NOR decondensation and activation was characterised by the formation of CMA+ blocks that resembled a string of beads inside the nucleolus. The number and size of beads was inversely proportional to the size of the nucleolus, suggesting that transcriptional activation of the nucleolar cistrons starts simultaneously at several points of the NOR. The mode of NOR activation in A. angustifolia differs from that observed in most species, providing a unique opportunity to study activation and transcriptional control of rRNA genes.

A supernumerary chromosome segment in Locusta migratoria

A single female of Locusta migratoria was found to be heterozygous for a supernumerary heterochromatic segment distally located on the M6 autosome close to its nucleolus organiser region (NOR). Reactions to several chromosome banding techniques revealed its heterochromatic nature and its composition of GC-rich DNA sequences and likewise the NORs in this species. This suggests an origin for the extra segment by amplification of GC-rich DNA sequences contained in the distal NOR of the M6 chromosome, which is reinforced by the observation that the NOR of segmented M6 chromosomes produced the larger nucleolus in embryo prophase cells, such as would be expected from the presence of rRNA genes in the extra segment. No accumulation mechanism was detected in this female after analyzing the 213 embryo offspring produced, but an increase in the number of nucleoli per interphase nucleus was noted in heterozygous embryos in respect to standard homozygous ones.Key words: Locusta migratoria, supernumerary segments, nucleolar organizing regions, heterochromatin.

Clustering of rDNA containing type 1 insertion sequence in the distal nucleolus organiser of Drosophila melanogaster: implications for the evolution of X and Y rDNA arrays

SummaryThe ribosomal RNAs produced by the multigene families on the X and Y chromosomes of Drosophila melanogaster are very similar despite the apparent evolutionary isolation of the X and Y chromosomal rDNA. X–Y exchange through the rDNA is one mechanism that may promote co-evolution of the two gene clusters by transferring Y rDNA copies to the X chromosome. This hypothesis predicts that the proximal rDNA of X chromosomes will be Y-like. Consequently, rDNA variants found only on the X chromosome (such as those interrupted by type 1 insertions) should be significantly clustered in the distal X nucleolus organizer. Proximal and distal portions of the X chromosome nucleolus organizer were separated by recombination between the inverted chromosomes In(1)scv2 (breakpoint in the centre of the rDNA) and In(1)sc4Lsc8R (no rDNA). Molecular analyses of the resulting stocks demonstrated that rRNA genes containing type 1 insertions were predominantly located on the chromosome carrying the distal portion of the X rDNA, thus confirming a prediction of the X–Y exchange hypothesis for the co-evolution of X and Y chromosomal rDNA. Distal clustering is not predicted by the alternative hypotheses of selection or gene conversion.