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Specific nuclear elimination in polyploid plasmodia of the slime mold Physarum polycephalum.

In growing plasmodia of the myxomycete Physarum polycephalum (G2-phase), three distinct classes of nuclei with a relative DNA content of 1x, 2x, and 4x are observed in the presumed haploid strain CL. The 2x and 4x species comprise up to 35% and 5% of the nuclei. Quantitative cytofluorometric studies of nuclei isolated in either G2- or S-phase or after FUDR treatment (G1 arrest) show that the three nuclear populations undergo a synchronous mitotic cycle and that the relative DNA content of the nuclear fractions in G-2 phase reflects the 2c, 4c, and 8c state. The heterogeneity of the nuclear population does, however, seem to be restricted to the growth phase. During a starvation period of 4 days that always preceeds sporulation (and also meiosis), the 4c nuclear population is reduced to 7%, 8c nuclei are no longer detected. These results suggest that a mechanism exists in Physarum for the selective detection and elimination of polyploid nuclei.

Cell Nucleus

Control of DNA synthesis in polyploid mammalian cells.

To test the hypothesis that the duration of DNA synthesis is an inverse function of nuclear size or DNA content, the S phase was calculated from PLM analysis for pseudodiploid, tetraploid, and octaploid lines of Chinese hamster cells growing as a monolayer or in suspension. S phase times were found not to be significantly different between polyploid lines and the diploid lines from which they were derived, regardless of the conformation of the nucleus. There is no evidence, therefore, that would implicate the nuclear membrane, or nuclear surface area/volume relationships, in the control of DNA synthesis.

Animals

Mitotically unstable polyploids in the yeast Pichia guilliermondii.

Attempts to obtain triploids or tetraploids of P. guilliermondii by sexual hybridization led to mitotically stable hybrids. However, their DNA content per cell was not higher than in diploids. The results of random spore analysis demonstrate that these hybrids were in fact aneuploids which obviously suffered drastic chromosome losses immediately after mating. This phenomenon could have been caused either by aneuploidy already present in the parental strains or it might have been due to a general inability of P. guilliermondii to maintain a polyploid genome.

Aneuploidy

Recombination between several polyploid loci.

The population genetics of polyploids classified at several loci studied in terms of the probabilistic mechanism of crossing over and segregation. The methods of genetic algebra are used.

Genetic Linkage

Chromosome organisation in polyploid mouse trophoblast nuclei.

At least one-third of mouse trophoblast cells undergo endoreduplication during the first half of gestation. It has been suggested that the endoreduplicated chromosomes may be polytenised. Here it is shown, using in situ hybridisation to the alpha-1 antitrypsin genes, which map at a unique site, that while there is a tendency for duplicated chromosomes to cluster, this does not involve the complete fusion of replicated chromatids found in fully polytene chromosomes, and in a substantial proportion of homologues the sites on the chromosome arms corresponding to these genes are widely separated. The centromeres do not fuse into a single chromocentre but the possibility is not ruled out that individual chromosomes may be polytenised in the centromeric region. Evidence is also presented showing that endoreduplication in trophoblast nuclei is not accompanied by the formation of new prekinetochore structures, in contrast to the situation in polyploid mouse liver and C127 cells.

Animals

Proportional polyploidization of 5S RNA genes in the ovary of Drosophila melanogaster mutants containing three 5S RNA gene loci.

The 5S RNA gene content of polyploid cells of the ovary of Drosophila melanogaster has been compared in animals with two or three gene clusters. The amount of 5S RNA genes is exactly proportional to the number of gene clusters as determined by DNA-RNA filter hybridization. In contrast, the number of rDNA genes in endomitotic cells remains constant regardless of different numbers of nucleolus organizer regions (Spear, 1974).

Animals

Polyploidization of G2M phase cells separated from aerobically and anaerobically grown Ehrlich ascites tumor cells.

G2-enriched fractions of Ehrlich ascites tumor cells (up to 80%-85% G2 cells) separated from anaerobically and aerobically cultured asynchronous populations by centrifugal elutriation revealed the same growth characteristics after recultivation under standard conditions: a significant proportion of cells with increased DNA (DNA content greater than 4C) emerged. Interruption of DNA synthesis by deprivation of oxygen may account for polyploidization (over-replication) of DNA but other mechanisms must be taken into consideration.

Aerobiosis

Focal necroses, fatty degeneration and subendocardial nuclear polyploidization of the myocardium in newborns after beta-sympathicomimetic suppression of premature labor.

It has been well documented in laboratory animals that beta-sympathicomimetics, such as isoprenalin, can cause myocardial lesions. Other so called "beta2-selective" symphaticomimetic drugs, which nevertheless induce beta1-cardiostimulatory side effects, are now widely used for suppression of premature labor. We examined the hearts of 25 newborns whose mothers had been treated with beta-sympathicomimetics for various lengths of time (24h to 8 weeks). Three types of lesions were detected: (1) focal subendocardial necroses (3 cases), similar to isoprenalin-induced myocardial necroses in animal experiments, (2) diffuse fatty degeneration of myocardial cells (3 cases), and (3) nuclear polyploidization in the subendocardial layer of the right ventricular wall (14 cases). However, the immediate causes of death could not be directly related to the tocolytic treatment in any of the cases.

Adrenergic beta-Agonists

Polyploidization and localisation of poly(A)+ RNA in the different cell types of the vitellogenic meroistic ovary of the fleshfly, Sarcophaga bullata.

The degree of polyploidization, the level of transcriptional activity and the volume of the different cell types present in the meroistic ovary of Sarcophaga bullata were measured during different vitellogenic stages. The nurse cells and the germinal vesicle exhibited very pronounced differences with regard to DNA content and mRNA synthesis, even though they are genetically identical. During the 4C stage (late vitellogenesis), we observed different degrees of polyploidy in follicle cells adjacent to the oocyte and those surrounding the nurse cells. Although the chromatin of the germinal vesicle is condensed into a karyosome, in situ hybridisation revealed the presence of transcriptional activity. The volume of the germinal vesicle, which contains only 4C DNA, is big enough to contain 2048C DNA. The meroistic ovary is a highly polarized differentiating system. Our results are discussed in the light of the fact that the polytrophic ovary is a miniature electrophoresis chamber.

Animals

Genome-wide cyclin gene evolution in Arabidopsis and Brassica reveals polyploidization-driven duplication and flowering-time associations.

Cyclin genes are plant cell cycle regulators that play essential roles in growth, development, and reproduction. However, the evolutionary dynamics and genomic organization of cyclin genes across the Brassicaceae family remain poorly understood, particularly in the context of allotetraploid genome evolution. Here, we investigated the diversity, expansion mechanisms, and potential functional diversification of cyclin genes across ten Brassicaceae genomes, including four Arabidopsis and six Brassica species. A total of 1087 cyclin genes representing 23 cyclin types were identified. Comparative genomic analyses revealed that cyclin gene expansion was strongly influenced by polyploidization in Brassica species, with 1845 duplication events involving 1063 genes. Whole-genome duplication was the predominant mechanism driving expansion, while both inter- and intra-genomic duplications contributed to gene retention in tetraploid Brassica species, with the highest duplication frequency observed in Brassica juncea. Across genomes, 120 physical gene clusters were identified, including homogeneous and heterogeneous types. Ortholog analysis between progenitor and allotetraploid species identified 852 orthologous pairs involving 366 genes, indicating extensive conservation following allotetraploid formation. Phylogenetic analysis resolved cyclins into three major clades, while expression-based clustering in Brassica napus grouped genes into four major clusters, suggesting functional diversification. Integration of pan-genomic and flowering-time QTL analyses further identified two cyclin genes, Bna21cycA2 and Bna113cycD4, which contain amino acid polymorphisms and represent putative candidate variations potentially associated with flowering-time variation across multiple genomes. These findings provide new insights into the evolutionary expansion, retention, and potential functional divergence of cyclin genes in Brassicaceae and highlight candidate loci for future functional studies and crop improvement.

Evolution, Molecular

Angiotensin evokes in polyploid rat glioma cells hyperpolarization-depolarization responses and cross-desensitization with bradykinin.

Angiotensins I, II and III induced a hyperpolarizing response of up to 1 min duration followed by a depolarizing response of up to 4 min when applied by pressure pulses or iontophoresis to polyploid rat glioma cells C6-4-2. The hyperpolarization (depolarization) was associated with a 50% decrease (no measurable change) in membrane resistance. The reversal potential (ca.-90 mV) of the hyperpolarization most likely points to an increase K+ conductance. Cells desensitized to angiotensins on application of high doses of either angiotensins or bradykinin.

Angiotensin II

A general treatment of chromosome synapsis in even-numbered polyploids.

Relationships between frequencies of fully paired pachytene configurations are examined for even-numbered polyploids, up to decaploids and beyond. Sequential association is assumed at any given number of equidistant synaptic sites showing uniform propensities for partner exchange. Only with two sites per chromosome, can each configuration represent an equal proportion of the nucleus. In the limit as the number of sites tends to infinity and the intervals between them tend to zero, pachytene multivalent frequency is seen to be a function of bivalent frequency, the minimum number of changes in pairing partner and ploidy. Each type of multivalent may represent the entire nucleus only in the lowest ploidy which can support it, thereafter becoming rare. Sets of homologous multivalents are never likely to be common and large multivalents are expected to be scarce, unless representing maximal configurations. Only bivalents can prevail at every even-numbered ploidy level. At constant bivalent frequency, the transition from discrete to continuous pairing favours the largest type of multivalent. These deductions provide a basis for investigations of the mechanics of chromosome synapsis and may lead to a comprehensive theory of genome analysis.

Cell Nucleus

Cytogenetic and molecular approaches of polyploidization in colorectal adenocarcinomas.

We present the cytogenetic analysis of 23 cases of polyploid colorectal adenocarcinomas. We took advantage of the high intratumoral heterogeneity of the karyotypes to identify clones, subclones, and cell-to-cell variations. This allowed us to reconstruct the chromosomal evolution of each tumor and to propose a schema of the chromosomal changes in relation to the endoreduplication process. All but one case were characterized by a relative deficiency of chromosomes 17p and 18. Other deficiencies affecting the late-replicating X, and to a lesser degree, 1p, 5q, 14, 15, 8p, 10, 21, and 4, and excesses affecting the early-replicating X, 8q, 13, 16, 17q, and 11 were frequently associated. This pattern of imbalances is very similar to that of the monosomic type previously described in near-diploid tumors. The pattern of the 23rd tumor corresponded to those of the trisomic type tumors. These data largely confirm the existence of two distinct processes of chromosomal evolution in colorectal adenocarcinomas, with a strong tendency to undergo endoreduplication for the monosomic type near-diploid tumors. To correlate cytogenetic and molecular data, allelic losses analyses were investigated for probes of chromosomes 17p and 18. In all 12 informative tumors, a loss of heterozygosity for probes of the short arm of chromosome 17 indicated the occurrence of a rearrangement of chromosome 17 before the endoreduplication. The same was true for allelic losses for probes of chromosome 18 found in 11 of 12 informative tumors. The correlation between cytogenetic and molecular data is thus excellent and indicates that losses of 17p and 18 are early events in the tumor process.

Adenocarcinoma

Metabolic utilization of diets by polyploid rainbow trout (Salmo gairdneri).

1. Nitrogen (NH4) excretion and oxygen consumption were measured in four groups of juvenile rainbow trout (36-40 g): triploids obtained by a heat shock treatment (3n); triploids obtained by mating diploid females with tetraploid males (3n/4n); tetraploids (4n) and a diploid control (2n). 2. No differences in daily N excretion patterns and in N excretion rate measured as a percentage of N intake were detected between groups. 3. Oxygen consumption of the 2n group was significantly inferior to that of the 3n and 4n groups but not to the 3n/4n group. 4. SDA, specific dynamic action (measured as a percentage of the energy intake), ranged from 5.6 to 8.6% and was not different between groups. 5. Protein energy catabolism measured as per cent of total energy expenditure was significantly higher in the 2n group than in polyploid groups.

Ammonia

Ribosomal RNA structure in the diploid and phylogenetically polyploid amphibian species Hyla and Odontophrynus.

Ribosomal RNA of the diploid amphibian species Hyla chrysoscelis and Odontophrynus americanus is structurally modified by hidden breaks. Phylogenetically polyploid related species like the tetraploid Hyla versicolor, the tetraploid Odontophrynus americanus and the octoploid Ceratophrys ornata do not show hidden breaks in ribosomal RNA. Structural modifications of rRNA molecules in diploid amphibians has no detectable effect on the ribosomal activity in vitro.

Amphibians

Transposable element-driven expansion of enhancer RNA repertoires underlies regulatory innovation and polyploid adaptation in cereal crops.

Cereal genomes have undergone repeated polyploidization and transposable element (TE) proliferation, collectively generating complex regulatory landscapes. However, the evolutionary trajectories and functional implications of these landscapes remain largely unexplored. Using chromatin-bound RNA sequencing across seven cereal species, we systematically mapped 45,952 regulatory element transcripts (RETs), including 32,867 distal RETs corresponding to enhancer RNAs (eRNAs). Our analysis revealed that 56% of lineage-specific eRNAs originated from TE expansions, indicating that TEs serve as major reservoirs of species-specific regulatory innovation in cereals. Notably, we identified remarkable conservation in defense-related functions, root-specific expression, and TE-derived origins of eRNAs across both ancient and recent evolutionary layers of Triticeae, suggesting recurrent recruitment of TE-derived, root-associated regulatory elements throughout Triticeae evolution. Furthermore, we found that young eRNA pairs in hexaploid wheat with high sequence similarity, many originating from RLG_famc8.3 and DTC_famc4.3, exhibited pronounced root specificity and coordinated expression, suggesting targeted amplification and refinement of successful ancestral regulatory strategies established after Triticeae divergence. To facilitate community access, we developed Cereal-eRNAdb (http://bioinfo.cemps.ac.cn/Cereal-eRNAdb/), a comprehensive database integrating 69,426 eRNAs with functional annotations across 296 samples. Our findings suggest that TE-mediated innovation of root-specific eRNAs may contribute to Triticeae adaptation and provide a foundational resource for exploiting regulatory variation in cereal crop breeding.

Enhancer RNAs

Comparative transmission genetics of introgressed chromatin in reciprocal advanced backcross populations in Gossypium (cotton) polyploids.

Introgression is a potential source of valuable genetic variation and interspecific introgression lines are important resources for plant breeders to access novel alleles. Experimental advanced-generation backcross populations contain individuals with genomic compositions similar to those resulting from natural interspecific hybridization and provide opportunities to study the nature and transmission pattern of donor chromatin in recipient genomes. Here, we analyze transmission of donor chromatin in reciprocal backcrosses between G. hirsutum and G. barbadense. Across the genome, recurrent backcrossing in both backgrounds yielded donor chromatin at slightly higher frequencies than the Mendelian expectation in BC5F1 plants, while the average frequency of donor alleles in BC5F2 segregating families was less than expected. In the two subgenomes of polyploid cotton, the rate of donor chromatin introgression was similar. Although donor chromatin was tolerated over much of the recipient genomes, 21 regions recalcitrant to donor alleles were identified. Only limited correspondence is observed between the recalcitrant regions in the two backgrounds, suggesting the effect of species background on introgression of donor segments. Genetic breakdown was progressive, with floral abscission and seed inviability ongoing during backcrossing cycles. Regions of either high or low introgression tended to be in terminal chromosomal regions that are generally rich in both genes and crossover events, with long stretches around the centromere having limited crossover activity resulting in relatively constant low introgression frequencies. Constraints on fixation and selection of donor alleles highlights the challenges of utilizing introgression breeding in crop improvement.

Humans

Reduced proliferative activity of polyploid cells in primary hepatocellular carcinoma.

The proliferative activity of tumor cells differing in DNA content (ploidy) and nuclearity was investigated in primary hepatocellular carcinomas of the rat by flow cytometric analysis of collagenase-isolated cells immunostained after labelling with bromodeoxyuridine (BrdU) in vivo. The diploid cell fraction in these euploid tumours was higher than in normal liver, and the rate of binucleation as well as the proliferative activity of the binuclear cells was very low. The highest proliferative activity (BrdU labelling index) was found among the diploid tumour cells. The activity in mononuclear tetraploid and octoploid cells was reduced in inverse proportion to their increasing DNA content, possibly suggesting a loss of proliferative potential associated with polyploidization. There was a significant correlation between the proliferative activity of hepatocellular carcinoma cells and nonparenchymal liver cells in the different tumours, indicating that different cell types within a tumour may respond to common growth stimuli. Treatment of tumour-bearing rats with a promoting carcinogen (2-acetylaminofluorene) resulted in significant stimulation of tumour cell proliferation (all ploidy classes), whereas the proliferation of non-parenchymal (stromal) cells in the tumour was slightly inhibited.

2-Acetylaminofluorene