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Nucleocytoplasmic ratio of fully grown germinal vesicle oocytes is essential for mouse meiotic chromosome segregation and alignment, spindle shape and early embryonic development.

BACKGROUND: This study examined the effect of nucleocytoplasmic ratio of fully grown germinal vesicle (GV) oocytes on meiotic chromosome segregation and alignment, spindle shape, Ca(2+) oscillations and capacity of early embryonic development in mouse. METHODS: GV oocytes with reduced volume (equal to 1/5 to 4/5 of an intact oocyte) were made by micromanipulation to remove different amounts of cytoplasm, and then matured and fertilized in vitro. RESULTS: When >1/2 of GV oocyte cytoplasm was removed, the time-course of GV breakdown (GVBD) was delayed and oocyte maturation rate decreased significantly. Abnormal chromosome segregation rate increased if >1/2 of the cytoplasm was removed from the oocyte. Length and structure of meiotic spindle and chromosome alignment were also impaired by the reduction of cytoplasmic volume. Once matured in vitro, the oocytes could undergo Sr(2+)-induced Ca(2+) oscillations and form pronuclei in a manner independent of nucleocytoplasmic ratio, but their ability to develop to 2-cell embryos was affected if >1/2 of their cytoplasm was removed from the GV oocytes. CONCLUSIONS: These results suggest that nucleocytoplasmic ratio is essential for normal meiotic chromosome segregation, spindle formation and chromosome alignment over the metaphase spindle, and development to 2-cell stage, for which 1/2 of the volume of the GV oocyte appears to be a threshold.

Animals↗

Meiotic segregation analysis in spermatozoa of pericentric inversion carriers using fluorescence in-situ hybridization.

BACKGROUND: Pericentric inversions are structural chromosomal abnormalities resulting from two breaks, one on either side of the centromere, within the same chromosome, followed by 180 degrees rotation and reunion of the inverted segment. They can perturb spermatogenesis and lead to the production of unbalanced gametes through the formation of an inversion loop. METHODS: We report here the analysis of the meiotic segregation in spermatozoa from six pericentric inversion carriers by multicolour fluorescence in-situ hybridization (FISH) and review the literature. RESULTS: The frequencies of the non-recombinant products (inversion or normal chromosomes) were 80% for the inv(20), 91.41% for the inv(12), 99.43% for the inv(2), 68.12% for the inv(1), 97% for the inv(8)(p12q21) and 60.94% for the inv(8)(p12q24.1). The meiotic segregation of 20 pericentric inversions (including ours) is now available. The frequency of unbalanced spermatozoa varies from 0 to 37.85%. The probability of a crossover within the inverted segment is affected by the chromosome and region involved, the length of the inverted segment and the location of the breakpoints. CONCLUSIONS: No recombinant chromosomes were produced when the inverted segment involved <30% of the chromosome length (independent of the size of the inverted segment). Between 30 and 50%, few recombinant chromosomes were produced, inducing a slightly increased risk of aneusomy of recombination in the offspring. The risk of aneusomy became very important when the inverted segment was >50% of the chromosome length. Studies on spermatozoa from inversion carriers help in the comprehension of the mechanisms of meiotic segregation. They should be integrated in the genetic exploration of the infertile men to give them a personalized risk assessment of unbalanced spermatozoa.

Adult↗

Segregation analysis of breast cancer from the cancer and steroid hormone study: histologic subtypes.

The segregation pattern of breast cancer in white families from the Cancer and Steroid Hormone Study was investigated. Families were categorized into four groups based on the histologic type of breast cancer in the probands:ductal cancer, lobular cancer, adenocarcinoma, and medullary cancer. The ductal cancer sample was further split into a premenopausal-proband and a postmenopausal-proband subset. Results for six complex segregation analyses are presented; the findings suggest heterogeneity in the transmission of breast cancer. For all analyses, there was no evidence for a multifactorial component in the mixed model, ie, a major locus plus other transmission, genetic and/or cultural. Interpretation of the medullary cancer, adenocarcinoma, and lobular cancer analyses does not permit discrimination among the major locus models. Segregation of breast cancer in the entire ductal sample was consistent with autosomal recessive transmission. In the ductal subanalyses, a recessive gene was sufficient to explain the breast cancer distribution when the proband had postmenopausal breast cancer. In contrast, when the proband had premenopausal breast cancer, the transmission model was consistent with a dominant major gene, with sporadic cases of disease.

Adenocarcinoma↗

On the mechanism of mitotic segregation induction in Aspergillus nidulans by benzene hydroxy metabolites.

The principal hydroxy-metabolites of benzene--hydroquinone, catechol and phenol--were assayed in tests for mitotic segregation induction in Aspergillus nidulans diploid strain 19. Hydroquinone was the most effective chemical, increasing the frequency of mitotic segregants up to 10-fold at 1-3 mM. Catechol was similarly active at 10-20 mM and phenol was weakly positive at 15 mM. Genetic characterization of induced abnormal segregating colonies by replating and complementary assays with haploid strain 35 suggest that gross chromosomal aberrations, instead of numerical abnormalities, are the primary genetic damages induced by hydroxybenzenes in A. nidulans. The protecting activity exerted by L-cysteine against equimolar concentrations of hydroquinone supports a free radical mechanism for hydroxybenzenes genotoxicity in A. nidulans.

Aspergillus nidulans↗

Fission yeast genes which disrupt mitotic chromosome segregation when overexpressed.

An interference assay has been devised in Schizosaccharomyces pombe to rapidly identify and clone genes involved in chromosome segregation. Random S.pombe cDNAs were overexpressed from an inducible promoter in a strain carrying an additional, non-essential minichromosome. Overexpression of cDNAs derived from four genes, two known (nda3+and ubc4+, encoding beta-tubulin and a ubiquitin conjugating enzyme, respectively) and two unknown, named mlo2+ and mlo3+ (missegregation & lethal when over expressed) caused phenotypes consistent with a failure to segregate chromosomes. Full overexpression of all four cDNAs was lethal. Cells overexpressing nda3+ and ubc4+ cDNAs arrested with condensed unsegregated chromosomes and cells overexpressing mlo2+ displayed an asymmetric distribution of nuclear chromatin. Sublethal levels of overexpression of nda3+, ubc4+ and mlo2+ cDNAs caused elevated rates of minichromosome loss. A third cDNA mlo3+, displayed no increase in the frequency of minichromosome loss at sublethal levels of overexpression but full overexpression caused a complete failure to segregate chromosomes. Our results confirm the assumption that beta-tubulin overexpression is lethal in S.pombe, implicate ubc4+ in the control of metaphase-anaphase transition in fission yeast and finally identify two new genes, mlo2+and mlo3+, likely to play an important role for chromosome transmission fidelity in mitosis.

Chromosomes, Fungal↗

Segregation analysis of prostate cancer in Sweden: support for dominant inheritance.

Several epidemiologic studies have shown familial aggregation of prostate cancer. To assess the nature of familial clustering of prostate cancer, a complex segregation analysis was performed on a population-based sample of 2,857 nuclear families ascertained through an affected father diagnosed with prostate cancer in Sweden during 1959-1963. The segregation analysis, using a large, unselected population of prostate cancer families, revealed that the observed clustering of prostate cancer was best explained by a high risk allele inherited in a dominant mode, with a high population frequency (1.67%) and a moderate lifetime penetrance (63%). This study confirms the result from one earlier published segregation analysis and provides the context for interpreting the recently published linkage of hereditary prostate cancer families to chromosome 1q 24-25 (HPC1).

Adult↗

The social adjustment of deaf adolescents in segregated parially integrated and mainstreamed settings.

This study examined the social adjustment of deaf adolescents enrolled in segregated (&egr; = 39), partially integrated (&egr; = 15), and mainstreamed (&egr; = 17) settings, comparing them with a control group of hearing students (&egr; = 88). Segregated students showed the lowest levels of adjustment overall. Partially integrated students reported better adjustment overall. Partially integrated students reported better adjustment than mainstreamed students with deaf peers; mainstreamed students reported better adjustment than partially integrated students with hearing peers, showing the same levels of adjustment with hearing peers as hearing students. Regardless of placement, deaf students reported better or equal adjustment with deaf than with hearing peers. Social adjustment with deaf peers was related to American Sign Language (ASL) skill and adjustment with hearing peers to spoken English. These findings suggest tht deaf students can benefit from both segregated and integrated placements as complememtary forms of social experience that each contribute to overal adjustment.

Journal Article↗

On the grain boundary segregation of Sn in indium-tin-oxide thin films

The determination of Sn segregation by means of electron energy loss spectroscopy and X-ray photoelectron spectroscopy has been attempted on indium-tin-oxide thin films prepared by various processes: (a) as-deposited at room temperature; (b) after annealing at 533 K in vacuum; (c) after annealing in air; and (d) as-deposited at 473 K. The occurrence of Sn segregation both at the surface and at the grain boundary was proved. The degree of segregation detected, varied depending on the preparation process of the films.

Journal Article↗

OCAM reveals segregated mitral/tufted cell pathways in developing accessory olfactory bulb.

Two functional subsets of vomeronasal sensory neurons project their axons to two segregated zones in the accessory olfactory bulb (AOB). Using immunohistochemical methods with antibodies against the novel cell adhesion molecule OCAM, we provide evidence that the segregation of functional pathways is maintained at the level of mitral/tufted (M/T) cells of the mouse AOB and that this pattern emerges early in ontogeny. During embryonic and postnatal development OCAM was strongly expressed by M/T cells in the caudal zone of the AOB where OCAM-negative vomeronasal axons terminated. In contrast, rostral zone M/T cells innervated by OCAM-positive vomeronasal axons displayed no or faint OCAM immunoreactivity. Differential expression of OCAM in segregated M/T cell pathways suggests that OCAM may be involved in defining compartments of connectivity and setting up functional subdivisions in the developing AOB.

Aging↗

Auditory stream segregation relying on timbre involves left auditory cortex.

An important aspect of auditory scene analysis is sequential grouping of sounds that are similar to one another in preference to sounds that follow one another. This grouping problem is captured by stream segregation tasks with alternating distinct sounds. We examined human auditory cortex activity with low noise fMRI in a stream segregation experiment relying on timbre differences of alternating harmonic tones (organ-like and trumpet-like). We found that stream segregation performance in comparison to monitoring a non-separable control stream increased activation exclusively in left auditory cortex and particularly in posterior areas. Our results suggest that left auditory cortex is selectively involved in this complex sequential task although the available cue for sequential grouping was timbre, usually attributed to right hemisphere analysis.

Adult↗

The mating consequences of sexual segregation within inflorescences of flowering plants.

Many co-sexual plants segregate female and male function among flowers on an inflorescence through dichogamy or the production of unisexual flowers. Sexual segregation may reduce self-pollination among flowers within inflorescences (geitonogamy), thereby increasing the pollen available for export to other plants. To assess these complementary roles we manipulated the simultaneously hermaphroditic (adichogamous) flowers of Eichhornia paniculata to produce ten-flowered inflorescences with either female above male flowers (female/male inflorescences) or male/female inflorescences, which competed for mating opportunities with five-flowered adichogamous inflorescences. Because of the upward movement of bumble-bees, selfing increased upward in adichogamous inflorescences (overall female selfing rate s+/-s.e.=0.320+/-0.026). Female flowers of male/female inflorescences selfed less than flowers in corresponding positions in adichogamous inflorescences so s fell to 0.135+/-0.027. In contrast, all-female flowers of female/male inflorescences selfed similarly to upper flowers on adichogamous inflorescences, elevating s (0.437+/-0.043). During 1997, male/female inflorescences sired more outcrossed seeds than female/male or adichogamous inflorescences, whereas during 1994 flowers on male/female inflorescences received fewer visits than those of adichogamous inflorescences, reducing their outcross siring success. Hence, sexual segregation limits geitonogamy and enhances outcross siring success when it does not affect pollinator behaviour, illustrating the importance of both female and male function in inflorescence design.

Animals↗

The segregation and integration of colour in motion processing revealed by motion after-effects.

Analysis of the colour and motion of objects is widely believed to take place within segregated processing pathways in the primate visual system. However, it is apparent that this segregation cannot remain absolute and that there must be some capacity for integration across these sub-modalities. In this study, we have assessed the extent to which colour constitutes a separable entity in human motion processing by measuring the chromatic selectivity of two kinds of after-effect resulting from motion adaptation. First, the traditional motion after-effect, where prolonged inspection of a unidirectional moving stimulus results in illusory motion in the opposite direction, was found to exhibit a high degree of chromatic selectivity. The second type of after-effect, in which motion adaptation induces misperceptions in the spatial position of stationary objects, was completely insensitive to chromatic composition. This dissociation between the chromatic selectivities of these after-effects shows that chromatic inputs remain segregated at early stages of motion analysis, while at higher levels of cortical processing there is integration across chromatic, as well as achromatic inputs, to produce a unified perceptual output.

Adaptation, Physiological↗

Sexual segregation of seasonal foraging habitats in a non-migratory marine mammal.

Many animal species segregate by sex. Such segregation may be social in nature, or ecological, or both. Grey seals (Halichoerus grypus), like many large mammals, are sexually size dimorphic. In size dimorphic species, allometric differences in morphology, metabolic rate and reproductive costs are likely. Such differences may require the sexes to use different foraging strategies or different habitats. To investigate sexual segregation of habitat in grey seals, we used satellite tracks from 95 (male 46; female 49) adults breeding at Sable Island, Nova Scotia (44 degrees N, 60 degrees W) collected from 1995 to 2005. Location estimates were made from satellite fixes using a state-space movement model to estimate true locations and regularize them in time. Location estimates were used to calculate home range kernels of male and female habitat use each month. Month by sex kernel home ranges revealed striking differences and dynamics in habitat use between males and females on spatial scales broader than most terrestrial examples and at temporal and spatial resolutions rarely available for marine species. Differences were most pronounced just before (October-December) and immediately after breeding (February-March). During both periods, males primarily used areas along the continental shelf break, while females mainly used mid-shelf regions. Coupled with previously identified sex-specific seasonal patterns of energy storage, diving and diet, our findings suggest that males and females differ profoundly in their spatial foraging strategies. These differences may serve to maximize fitness by reducing intersexual competition during key foraging periods.

Animal Migration↗

Auditory segregation of competing voices: absence of effects of FM or AM coherence.

Four experiments sought evidence that listeners can use coherent changes in the frequency or amplitude of harmonics to segregate concurrent vowels. Segregation was not helped by giving the harmonics of competing vowels different patterns of frequency or amplitude modulation. However, modulating the frequencies of the components of one vowel was beneficial when the other vowel was not modulated, provided that both vowels were composed of components placed randomly in frequency. In addition, staggering the onsets of the two vowels, so that the amplitude of one vowel increased abruptly while the amplitude of the other was stationary, was also beneficial. Thus, the results demonstrate that listeners can group changing harmonics and can segregate them from stationary harmonics, but cannot use coherence of change to separate two sets of changing harmonics.

Acoustic Stimulation↗

Loss of the toluene-xylene catabolic genes of TOL plasmid pWW0 during growth of Pseudomonas putida on benzoate is due to a selective growth advantage of 'cured' segregants.

During growth on benzoate-minimal medium Pseudomonas putida mt-2 (PaW1) segregates derivative ('cured') strains which have lost the ability to use the pathway encoded by its resident catabolic plasmid pWW0. Experiments with two plasmids identical to pWW0 but each with an insert of Tn401, which confers resistance to carbenicillin, suggested that the 'benzoate curing' occurs far more frequently by the specific deletion of the 39 kbp region carrying the catabolic genes than by total plasmid loss. This effect was not pH-dependent, and was not produced during growth on other weak organic acids, such as succinate or propionate, or when benzoate was present in the medium with an alternative, preferentially used carbon source such as succinate. Growth on benzoate did not cause loss from strain PaW174 of the plasmid pWW0174, a derivative of pWW0 which has deleted the 39 kbp region but carries Tn401. Similarly the naphthalene-catabolic plasmid pWW60-1, of the same incompatibility group as pWW0, was not lost from PaW701 during growth on benzoate. Competition between wild-type PaW1 and PaW174, which has the 'cured' phenotype, showed that the latter has a distinct growth advantage on benzoate over the wild-type even when initially present as only 1% of the population: when PaW174 was seeded at lower cell ratios, spontaneously 'cured' derivatives of PaW1 took over the culture after 60-80 generations, indicating that they are present in PaW1 cultures at frequencies between 10(-2) and 10(-3). We conclude that the progressive takeover of populations of PaW1 only occurs when benzoate is present as the sole growth source and that neither benzoate, nor other weak acids, affect plasmid segregation or deletion events: a sufficient explanation is that the 'cured' segregants grow faster than the wild-type using the chromosomally determined beta-ketoadipate pathway.

Benzoates↗

Paternal DNA strands segregate to both trophectoderm and inner cell mass of the developing mouse embryo.

The localization of sperm DNA strands was examined in preimplantation mouse embryos. Male mice were treated with bromodeoxyuridine (BrdU) to label germ-cell-line DNA and were then mated with unlabeled females. Sperm DNA strands in early embryos derived from these matings could be detected by means of a fluorescent antibody specific to BrdU. The position and number of fluorescent spots detected in the developing embryos are consistent with a model in which paternal DNA strands segregate at random into both the trophectoderm and the inner cell mass. Although we could not follow the segregation of individual paternal chromosomes, we could detect no overall segregation pattern of the sperm DNA strands that could be obviously related to chromosome imprinting.

Animals↗

Size segregation of granular matter in silo discharges.

We present an experimental study of segregation of granular matter in a quasi-two-dimensional silo emptying out of an orifice. Size separation is observed when multisized particles are used with the larger particles found in the center of the silo in the region of fastest flow. We use imaging to study the flow inside the silo and quantitatively measure the concentration profiles of bidisperse beads as a function of position and time. The angle of the surface is given by the angle of repose of the particles, and the flow occurs in a few layers only near the top of this inclined surface. The flowing region becomes deeper near the center of the silo and is confined to a parabolic region centered at the orifice which is approximately described by the kinematic model. The experimental evidence suggests that the segregation occurs on the surface and not in the flow deep inside the silo where velocity gradients also are present. We report the time development of the concentrations of the bidisperse particles as a function of size ratios, flow rate, and the ratio of initial mixture. The qualitative aspects of the observed phenomena may be explained by a void filling model of segregation.

Journal Article↗

Segregation of granular binary mixtures by a ratchet mechanism.

We report on a segregation scheme for granular binary mixtures, where the segregation is performed by a ratchet mechanism realized by a vertically shaken asymmetric sawtooth-shaped base in a quasi-two-dimensional box. We have studied this system by computer simulations and found that most binary mixtures can be segregated using an appropriately chosen ratchet, even when the particles in the two components have the same size and differ only in their normal restitution coefficient or friction coefficient. These results suggest that the components of otherwise nonsegregating granular mixtures may be separated using our method.

Journal Article↗