It is well known that two DNA molecules are wrapped around histone octamers and folded together to form a single chromosome. However, the nucleosome fiber folding within a chromosome remains an enigma, and the higher-order structure of chromosomes also is not understood. In this study, we employed electron diffraction which provides a noninvasive analysis to characterize the internal structure of chromosomes. The results revealed the presence of structures with 100–200 nm periodic features directionally perpendicular to the chromosome axis in unlabeled isolated human chromosomes. We also visualized the 100–200 nm periodic features perpendicular to the chromosome axis in an isolated chromosome whose DNA molecules were specifically labeled with OsO4 using electron tomography in 300 keV and 1 MeV transmission electron microscopes.

The spatial organization of chromatin has become recognized as a major factor in the regulation of gene expression. While chromosome conformation capture techniques, such as high-throughput chromosome conformation capture (Hi-C), have advanced our understanding of the role of 3D organization of chromatin on chromosomal function, Hi-C data are typically ensemble in nature and relies on intense computational methods to derive genomic structural information. The field is beginning to shift in recognizing the potential value of imaging methods to study spatial organization of chromatin. Imaging allows acquisition and analysis at the individual cell level while providing direct visualization and analysis of 3D chromatin organization. This feature will highlight how single-molecule localization microscopy (SMLM) using the Bruker Vutara SMLM system in conjunction with sequential labeling via microfluidics is revolutionizing our understanding of nuclear spatial organization and its role in regulation of gene function.

Approximately one-quarter of all herbicides that have been marketed affect mitosis as a primary mechanism of action. All of these herbicides appear to interact directly or indirectly with the microtubule. Dinitroaniline and phosphoric amide herbicides inhibit microtubule polymerization from free tubulin subunits. Because of the loss of spindle and kinetochore microtubules, chromosomes cannot move to the poles during mitosis, resulting in cells exhibiting an arrested prometaphase configuration. Nuclear membranes re-form around the chromosomal masses to form lobed nuclei. Cortical microtubules, which influence cell shape, are also absent, and, as a result, the cell expands isodiametrically. In root tips and other structures that are normally elongated, these herbicides induce a characteristic club-shaped swelling. Pronamide and MON 7200 induce similar effects, except that tufts of microtubules remain at the kinetochore region of the chromosomes. The carbamate herbicides barban, propham, and chlorpropham alter the organization of the spindle microtubules so that multiple spindles are formed. Chromosomes move to many poles and multiple nuclei result. Abnormal branched cell walls partly separate the nuclei. Terbutol induces “star anaphase” chromosome configurations in which the chromosomes are drawn into an area at the poles in a star-like aggregation. DCPA's most dramatic effect is on phragmoplast microtubule arrays. Multiple, branched, and curved phragmoplasts are found after herbicide treatment. These disrupters should prove to be useful tools in investigations of the proteins and structures required for a successful cell division.

Studies were conducted to identify and characterize different accessions of itchgrass. Seeds were collected in the counties of Aramina, Campinas, Dumont, Igarapava, Jaboticabal, and Ribeirão Preto, all in the state of São Paulo, Brazil. Accessions were characterized based on dimensions of their stomata, stomatal index (SI), and length and width of their seed (caryopses and husk). Chromosome number and length also were determined, and accessions were further differentiated using molecular markers (polymerase chain reaction [PCR]). Itchgrass from Ribeirão Preto had much longer and narrower seeds than those from the other locations, and their husks were longer as well. Accessions had similar SIs, both on the abaxial and adaxial leaf surfaces. Stomata from Campinas and Igarapava accessions were longer and wider, whereas those from Dumont and Ribeirão Preto were similar and smaller than all others. The accession from Ribeirão Preto is diploid (2n = 20); the rest are polyploid, with the total length of chromosomes smaller than all others. These differences were confirmed by molecular differentiation (PCR).

Chromosomes were counted for nine of the 24 taxa of Berberis (Berberidaceae) from Nepal, five of which were counted for the first time. The results show that all these species have somatic chromosome numbers of 2n = 28. The absence of polyploids suggests that polyploidy may not have been a major driver of speciation and diversification of Berberis in the Nepal Himalaya.

This paper explores evolving conceptions and depictions of the human genome among human and medical geneticists during the postwar period. Historians of science and medicine have shown significant interest in the use of informational approaches in postwar genetics, which treat the genome as an expansive digital data set composed of three billion DNA nucleotides. Since the 1950s, however, geneticists have largely interacted with the human genome at the microscopically visible level of chromosomes. Mindful of this, I examine the observational and representational approaches of postwar human and medical genetics. During the 1970s and 1980s, the genome increasingly came to be understood as, at once, a discrete part of the human anatomy and a standardised scientific object. This paper explores the role of influential medical geneticists in recasting the human genome as being a visible, tangible, and legible entity, which was highly relevant to traditional medical thinking and practice. I demonstrate how the human genome was established as an object amenable to laboratory and clinical research, and argue that the observational and representational approaches of postwar medical genetics reflect, more broadly, the interdisciplinary efforts underlying the development of contemporary biomedicine.

A dual cytogenetic and molecular analysis was performed in four species of Cyclocepala (Coleoptera: Scarabaeidae: Dynastinae) from Lesser Antilles (Martinique, Dominica and Guadeloupe). Two species/sub-species, C. mafaffa grandis and C. insulicola, are endemic to Guadeloupe. They have their own non-polymorphic karyotype and a fairly homogeneous haplotype of the COI gene. C. melanocephala rubiginosa has a distinct karyotype. Its COI haplotype is homogeneous in Guadeloupe and heterogeneous in Martinique. Finally, C. tridentata has highly different karyotypes and haplotypes in the three islands. In Martinique, its karyotype, composed of metacentrics, is monomorphic while its haplotype is fairly heterogeneous. Both are close to those of other Cyclocephala and Dynastinae species, thus fairly ancestral. In Guadeloupe, its karyotype is highly polymorphic, with many acrocentrics, and its haplotype fairly homogeneous. Both are highly derived. In Dominica, both the karyotype and the haplotype represent intermediate stages between those of Martinique and Guadeloupe. We conclude that several independent colonization episodes have occurred, which excludes that C. insulicola is a vicariant form of C. tridentata in Guadeloupe. Both chromosome and COI gene polymorphisms clearly indicate a recent colonization with a northward direction for C. tridentata.

In cockchafers of the genus Melolontha, there is a marked intraspecific polymorphism for morphological characters, making some specimens of one species resemble another. A cytogenetic and molecular (mitochondrial COI gene sequence) study of typical and atypical forms of M. melolontha and M. hippocastani, captured at the same period and area, was performed. Karyotypes and haplotypes clearly characterize each taxon, placing atypical specimens in one or the other species unambiguously. This formally discards the role of hybridization in phenotypic resemblance, as usually proposed. Karyotypes and haplotypes were compared to those of M. pectoralis and Phyllophaga pleei, a more distantly related Melolonthinae, and some Dynastinae species, to reconstruct their ancestral karyotype. The karyotype of M. melolontha is the most derivative and that of P. pleei the most conserved among the Melolonthinae studied, which fits with the phylogeny established by COI gene analysis. Both karyotypes and COI haplotypes demonstrate the proximity of M. pectoralis and M. melolontha. The karyotype of M. melolontha is polymorphic, without relationship with morphological variations. Finally, the existence of similar morphological variations in different Melolontha species and chromosomal polymorphism in M. melolontha is discussed in relation with a network (reticulated) mode of speciation.

Antarctic plunderfishes (Notothenioidei, Artedidraconidae) are important components of the Southern Ocean fish fauna. As a contribution to the Victoria Land Transect Project, we performed a cytogenetic analysis of six species from three of the four artedidraconid genera, Artedidraco glareobarbatus, A. orianae, A. skottsbergi, A. shackletoni, Histiodraco velifer, and Pogonophryne sp. We investigated the species-specific cytogenetic features and highlighted patterns of chromosomal evolutionary change using a molecular phylogeny based on mitochondrial and nuclear genes. Despite a conserved diploid number, some important karyotypic traits account for major differences among artedidraconid species. Specific cytogenetic features, including the chromosomal organization of ribosomal genes and the occurrence of sex chromosomes, are characteristics of A. skottsbergi, making this species distinct among those studied. These chromosomal peculiarities are consistent with the phylogenetic hypothesis resolving A. skottsbergi as the sister lineage of all other Artedidraconidae. A karyological similarity was found between A. glareobarbatus and A. shackletoni consistent with their inferred sister species relationship in the phylogeny. The results indicate that artedidraconids are not conservative in their genomic organization at the chromosomal level and provide new evidence for the degree of biological diversity in this notothenioid group.

Karyotypes for several East Pacific Rise hydrothermal vent invertebrates are described here for the first time: the vestimentiferans Riftia pachyptila and Oasisia alvinae, the alvinellid polychaetes Alvinella pompejana, A. caudata and Paralvinella grasslei, the polynoid polychaetes Branchinotogluma grasslei and Branchipolynoe symmytilida, the serpulid Laminatubus alvini and the mytilid bivalve Bathymodiolus thermophilus. For comparative purposes, the karyotype of the Atlantic vent mussel Bathymodiolus azoricus is also described here for the first time. Each species has its own unique chromosomal characteristics which can be interpreted both in terms of group characteristics and species divergence. From comparisons with published results on other vent species and closely-related coastal species, we identified a positive correlation between chromosome number variation and molecular divergence at two ribosomal ribonucleic acid gene loci (the 18S and 28S rRNA). Whilst the patterns of chromosome divergence we found were generally within the ranges previously reported for these taxonomic groupings, there was an apparent inconsistency in the case of Branchipolynoe symmytilida (EPR) and Branchipolynoe seepensis (MAR), which show a greater degree of divergence at the chromosome level compared with other members of the same genus. Moreover, polychaetes as a whole showed greater variation in the number and structural divergence of chromosomes compared to Mytilids (structural information only). Our findings highlight the great potential for chromosome analysis in future taxonomic and evolutionary studies of the deep-sea vent fauna.

The karyotype of Isthmiophora melis (Trematoda:Echinostomatidae) is described. The chromosome number in the diploid cell is 2n = 20. The absolute (La) and relative (Lr) lengths of the chromosomes, as well as the centromere position (Ic), are determined. The differences in length between the adjacent chromosomes in the ordered series are not great, with the exception of chromosome 1 which occupies 20.58% of the genome length. This chromosome is metacentric, with lightly stained and elongated centromere region with Giemsa staining. Chromosome 7 is submetacentric. Chromosomes 2 to 6 are defined as telo-subtelocentric, while chromosomes 8 to 10 are subtelo-submetacentric. The fundamental number of arms in the I. melis karyotype is NF-22.

Chromosomes of Nigerian Onchocerca volvulus were compared with those of Guatemalan O. volvulus. Both parasites had basically the same chromosomal construct (2n=8, XY type). Autosomes consisted of a pair of large and two smaller pairs. Sex chromosomes were made up of medium sized X chromosome and very small Y chromosome. It was not possible to infer the position of the centromeres.

Chromosomal complements of an oocyte and a fertilized ovum from the ootype of Isoparorchis hypselobagri Billet, 1898 have been studied. Twenty diploid and 10 haploid chromosomes have been observed. In the karyotype the presence of five submetacentric, four acentric and a pair of sex chromosomes provide a significant cytotaxonomic information.

Testicular examination of male Onchocerca dukei and German O. tarsicola revealed five pairs of chromosomes, which is typical of most filariae. In contrast to German O. tarsicola, specimens from Sweden had only three pairs of chromosomes.

The diploid chromosome number of Protostrongylus rufescens is 2n=11 for males and 2n=12 for females. So, the sex determinism mechanism is XO/XX. The study of the genetic behaviour of this species has been made. In diakinesis stage the bivalents show typical tetrads with cross, Ø, and lineal configurations. The division of the sexual chromosome is prereductional for the first meiotic division.

Introduction. Iran, with more than 10 species of Pyrus, is one of the important genetic resources for this genus in the world, and Fars province is one of the centers of origin. In our research, cytogenetical investigation of native wild pear species of Fars province, including P. glabra Boiss., P. syriaca Boiss. and, probably, a natural hybrid of them, (P. glabra × P  syriaca), was carried out using a video analysis system. Materials and methods. Seed germination of different species was done on damp filter paper in petri dishes, and root tips were collected for karyotypical investigation. After pretreatment, fixation, hydrolysis and staining, samples were prepared for observation under the microscope and chromosome morphology was studied. Analysis of the species genome (length of each chromosome, length of long and short arm, ratio of long arm to short arm and ratio of short arm to long arm) was done and the chromosomal type of each species was determined using Levan et al.'s method. Then the karyotypic symmetry of species was studied by Stebbins' method. Results and discussion. The results revealed that all of the genotypes, with 2n = 2x = 34, were diploid. Based on Stebbins' table, genotypes 1 and 2 of P. glabra were classified into group 1A, P. glabra × P. syriaca into group 2A and P. syriaca into group 2B. Genotype plots, according to the A1 and A2 parameters and Stebbins' cross classes, revealed the same results. Similarity and differences of species in chromosomal aspects were investigated and the results shown in a dendrogram.

Spindles were isolated from deposited eggs of the Mediterranean mealmoth, Ephestia Kuehniella. Their structure and development were studied using anti-tubulin immunofluorescence. The microtubules were labelled with three different monoclonal antibodies. These were directed against β-tubulin, tyrosinated α-tubulin and acetylated α-tubulin. Significant differences in the staining behaviour were not detected with the three antibodies. An unusual mode of spindle formation was observed during the first mitotic division after fusion of the pronuclei. Several of the ensuing embryonic divisions may show the same phenomenon. Prophase of these divisions was characterised by an irregular arrangement of microtubules in the nuclear area. The microtubule mass in the nuclear area increased concomitantly with chromosome condensation. Microtubular foci, comparable to the forming asters of canonical spindles, were not detected. The formation of an orderly pattern in the microtubule mass was signalled by the appearance of minispindles apparently developing around individual chromosomes. Several minispindles subsequently aligned and formed metaphase-like entities within the nuclear area. The metaphase-like entities, in turn, aligned with one another and gave rise to a conventional bipolar metaphase spindle with small asters. The further development of the spindle was conventional. The chromosomes migrated towards the spindle poles and finally daughter nuclei formed. The anaphase and telophase spindles possessed both a prominent array of interzone microtubules and asters. The events in prophase of early embryonic mitosis of E. kuehniella may represent a rare case of chromosomeinduced spindle formation.

Previously we have shown that mitogen-activated protein (MAP) kinase activity abruptly increases at the first metaphase (M1) and remains significantly higher than that at the germinal vesicle (GV) stage until the second metaphase (M2) in porcine oocytes cultured in vitro. The present paper describes how the mechanism of the blockage of meiotic maturation by protein sythesis inhibition involves MAP kinase regulation. Cycloheximide arrested both germinal vesicle breakdown (GVBD) and the normal transition from M1 to M2. MAP kinase activation was also reduced in these maturation-inhibited oocytes. By using immunofluorescence microscopy with the monoclonal antibody raised against rat α-tubulin, we showed that cycloheximide caused morphological abnormality in a spindle at M1, but not at M2. All these results indicate that in porcine oocytes: (1) GV blockage by protein synthesis inhibition involves the suppression of both histone H1 kinase and MAP kinase activation, (2) during the transition from M1 to M2, maintenance of a normal metaphasic spindle and high MAP kinase activity require protein synthesis, and (3) once the M2 cytoskeletal structures have been completed, and/or after the ‘critical period’, cytostatic factor activity is independent of protein synthesis.

Pretreatment of zona-free Chinese hamster (CH) oocytes with three kinds of lectin – concanavalin A (Con-A), phytohaemagglutinin-P (PHA) and wheat germ agglutinin (WGA) – was attempted in order to improve penetration by golden hamster (GH) spermatozoa in vitro. Con-A had no significant effect on penetration at 2 μg/ml, adequately facilitated oocyte-sperm fusion at 4 μg/ml, and caused excessive sperm binding and resultant severe polyspermy at 10 μg/ml. Neither PHA nor WGA had positive effects on sperm penetration at any concentrations (2–10 μg/ml) examined. Using the Con-A (4 μg/ml)pretreatment, high rates of interspecific fertilisation and subsequent chromosome analysis of hybrid 1-cell zygotes were achieved. Among 258 CH oocytes used, 212 (82.2%) were fertilised and 153 (72.2% of fertilised ova) developed to the first cleavage metaphase. Eventually, 132 CH-derived chromosome complements and 153 GH-derived ones were successfully karyoanalysed. Incidences of aneuploidy and structural anomaly were 3.1% and 2.3% in CH complements, and 1.4% and 6.5% in GH complements, respectively. These incidences were not significantly different from those obtained by intraspecific in vivo fertilisation, suggesting that our interspecific in vitro fertilisation system does not cause chromosome aberrations.