Academic literature on the topic 'Heteroploidia'

We found that salt water induces ploidy alterations in salmonid embryos. Flow cytometry analysis revealed significantly higher frequencies of haploids, triploids, heteroploid mosaics, and aneuploids in rainbow trout (Oncorhynchus mykiss) embryos experimentally exposed to salt water from fertilization to the two- and eight-cell stages of development. Heteroploid mosaics have been reported in diploid and triploid salmonid hybrids, although none were observed in the triploid coho salmon or diploid and triploid coho salmon (O. kisutch) × chinook salmon (O. tshawytscha) hybrids we examined. No mosaics were observed in intertidally spawned pink salmon (O. gorbuscha) embryos. Salt water could induce ploidy alterations by causing chromosome segregation errors during meiosis, mitosis, or both. Heteroploid embryos appeared morphologically normal although they may possess physiological deficiencies not immediately apparent.

In the present study, we aimed to check whether uterine leiomyomas (ULs) with an apparently normal karyotype in vitro comprise “hidden” cell subpopulations with numerical chromosome abnormalities (heteroploid cells). A total of 32 ULs obtained from 32 patients were analyzed in the study. Each UL was sampled for in vivo and in vitro cytogenetic studies. Karyotyping was performed on metaphase preparations from the cultured UL samples. A normal karyotype was revealed in 20 out of the 32 ULs, of which 9 were selected for further study based on the good quality of the interphase preparations. Then, using interphase FISH with centromeric DNA probes, we analyzed the copy number of chromosomes 7 and 16 in 1,000 uncultured and 1,000 cultured cells of each selected UL. All of the ULs included both disomic cells representing a predominant subpopulation and heteroploid cells reaching a maximum frequency of 21.6% (mean 9.8%) in vivo and 11.5% (mean 6.1%) in vitro. The spectrum of heteroploid cells was similar in vivo and in vitro and mostly consisted of monosomic and tetrasomic cells. However, their frequencies in the cultured samples differed from those in the uncultured ones: while the monosomic cells decreased in number, the tetrasomic cells became more numerous. The frequency of either monosomic or tetrasomic cells both in vivo and in vitro was not associated with the presence of <i>MED12</i> exon 2 mutations in the tumors. Our results suggest that ULs with an apparently normal karyotype consist of both karyotypically normal and heteroploid cells, implying that the occurrence of minor cell subpopulations with numerical chromosome abnormalities may be considered a characteristic of UL tumorigenesis. Different frequencies of heteroploid cells in vivo and in vitro suggest their dependence on microenvironmental conditions, thus providing a pathway for regulation of their propagation, which may be important for the UL pathogenesis.

Cytogenetic analysis of solid tumors with comparative genomic hybridization (CGH) is hampered by the dilution of DNA from individual tumor subpopulations with DNA from other cells. We investigated to what extent this dilution effect can be alleviated using fluorescence activated cell sorting (flow sorting) of experimental DNA heteroploid cell mixtures prior to CGH. From mixtures of normal lymphocytes with triploid K‐562 cells the individual components were sorted according to stemline DNA content and processed by CGH in comparison with pure K‐562 samples and the original mixtures. Compared with 30 autosome copy number imbalances found in pure K‐562 samples, a mixture with 32% K‐562 cells showed 16 imbalancies, and none were detected in mixtures with 13% or 5% K‐562 cells. In contrast, 29, 22 and 23 imbalances were detected in K‐562 nuclei sorted from the 32%, 13% and 5% mixtures, respectively. This indicate that CGH analysis of flow sorted DNA aneuploid subpopulations enables a specific cytogenetic analysis of the individual subclones in a DNA heteroploid cell population.

Orientador: Yoko Bomura Rosato<br>Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia<br>Made available in DSpace on 2018-07-15T09:03:12Z (GMT). No. of bitstreams: 1 Moscoso_IrmadeLourdes_M.pdf: 4592907 bytes, checksum: 7da4f18d51329da239213f41b89bdef9 (MD5) Previous issue date: 1987<br>Resumo: Linhagens haplóides, bem como heterocários e diplóides do fungo Aspergillus nidulans foram analisados quanto à produção das enzimas extracelulares lipase, amilase, protease, fosfatase e urease. Foram utilizados três linhagens normais e oito mutantes morfológicos, além de diferentes combinações entre ambos. Os ensaios foram efetuados em meios de cultura sólidos, contendo cada qual, um substrato especifico. As linhagens morfológicas do tipo compacto foram também analisadas microscopicamente, e comparadas às demais, previamente descritas. Pelos resultados obtidos, foi constatado que as colônias de crescimento reduzido em um ou mais meios de cultura foram sempre as mais alteradas também no aspecto enzimático, bem corno no microscópico. As compactas B-VIII e B-6, bem corno os heterocários pp+M-32 e pp+M-35, destacaram-se por apresentar a maior produção de lipase, amilase, protease, enquanto que outras linhagens e heterocários mostraram maior produção em relação a apenas uma ou outra dessas enzimas. A linhagem compacta pp-6 constituiu uma exceção, já que não apresentou produção de protease, embora o heterocário MSE + pp-6 tenha produzido o mais alto nível de atividade dessa mesma enzima. Para fosfatase e urease ocorreu o inverso, isto é, as colônias de menores dimensões apresentaram evidências de menor atividade enzimática. Quanto aos diplóides, houve restauração do fenótipo enzimático original bm ou pp em todos os casos. As alterações aqui descritas foram discutidas, considerando-se a possibilidade de problemas de permeabilidade nos mutantes morfológicos utilizados<br>Abstract: The production of extracellular enzynes such as lipase, amylase, protease, phosphatase and urease by haploids, heterocaryons and diploids of A. nidulans were analysed. Three normal strains, eight morphological mutants and differents combinations among them were utilized. The enzymatic test were carried out using solid culture medium containing specific substrate for each enzyme under assay. The results shwed that slow growing colonies in one or more culture medium showed also more alterations in the enzyme production. In general compact colonies (B-VIII and B-6) and some heterocaryons (pp+M-32 and pp+M-35) showed higher production of lipase, amylase and protease. For specific enzymes there are others strains and heterocaryons that must be considered. The strain pp-6 was a special compact strain showing no activity of protease at all; however in the heterocaryon MSE + pp-6 there was a strong interaction producing the highest level of protease activity. For phosphatase and urease the results were quite different, the smaller colonies having lower enzymatic activity. There were no differences in the diploids and the level of enzymes production returned to the respective original strain bm ou pp. All the alterations described, specially in haploids and heterocaryons were discussed considering permeability problems in the morphological mutants<br>Mestrado<br>Genetica<br>Mestre em Ciências Biológicas

The cell cycle is a highly complex process that is under the control of several pathways.  Failure to regulate and/or complete the cell cycle often leads to cell cycle arrest, which may be followed by programmed cell death (apoptosis). One cell type that has a variety of unique cell cycle properties is the horizontal cell of the chicken retina. In this thesis we aimed to characterize the final cell cycle of retinal horizontal cells. In addition, the regulation of the cell cycle and the resistance to apoptosis of retinal horizontal cells are investigated. Our results show that the final cell cycle of Lim1-expressing horizontal progenitor cells is heterogenic and three different cell cycle behaviors can be distinguished. The horizontal cells are generated by: (i) an interkinetic nuclear migration with an apical mitosis; (ii) a final cell cycle with an S-phase that is not followed by mitosis, such cells remain with a fully or partially replicated genome; or (iii) non-apical (basal) mitoses. Furthermore, we show that the DNA damage response pathway is not triggered during the heterogenic final cell cycle of horizontal progenitor cells. However, chemically induced DNA damage activated the DNA damage response pathway without leading to cell cycle arrest, and the horizontal progenitor cells entered mitosis in the presence of DNA damage. This was not followed by apoptosis, despite the horizontal cells being able to functionally activate p53, p21CIP1/waf1, and caspase-3. Finally, we show that FoxN4 is expressed in horizontal progenitor cells and is required for their generation. Over-expression of FoxN4 causes cell death in several neuronal retinal cell types, except horizontal cells, where it results in an overproduction. In conclusion, in this thesis, a novel cell cycle behavior, which includes endoreplication not caused by DNA damage and a basal mitosis that can proceed in the presence of DNA damage, is described. The cell cycle and cell survival processes are of particular interest since retinal horizontal cells are suggested to be the cell-of-origin for retinoblastoma.

Retinal progenitor cells undergo apical mitoses during the process of interkinetic nuclear migration and newly generated post-mitotic neurons migrate to their prospective retinal layer. Whereas this is valid for most types of retinal neurons, chicken horizontal cells are generated by delayed non-apical mitoses from dedicated progenitors. The regulation of such final cell cycle is not well understood and we have studied how Lim1 expressing horizontal progenitor cells (HPCs) exit the cell cycle. We have used markers for S-and G2/M-phase in combination with markers for cell cycle regulators Rb1, cyclin B1, cdc25C and p27Kip1 to characterise the final cell cycle of HPCs. The results show that Lim1+ HPCs are heterogenic with regards to when and during what phase they leave the final cell cycle. Not all horizontal cells were generated by a non-apical (basal) mitosis; instead, the HPCs exhibited three different behaviours during the final cell cycle. Thirty-five percent of the Lim1+ horizontal cells was estimated to be generated by non-apical mitoses. The other horizontal cells were either generated by an interkinetic nuclear migration with an apical mitosis or by a cell cycle with an S-phase that was not followed by any mitosis. Such cells remain with replicated DNA and may be regarded as somatic heteroploids. The observed heterogeneity of the final cell cycle was also seen in the expression of Rb1, cyclin B1, cdc25C and p27Kip1. Phosphorylated Rb1-Ser608 was restricted to the Lim1+ cells that entered S-phase while cyclin B1 and cdc25C were exclusively expressed in HPCs having a basal mitosis. Only HPCs that leave the cell cycle after an apical mitosis expressed p27Kip1. We speculate that the cell cycle heterogeneity with formation of heteroploid cells may present a cellular context that contributes to the suggested propensity of these cells to generate cancer when the retinoblastoma gene is mutated.

Desde el inicio de un cultivo celular, las células que lo conforman atraviesan una serie de presiones de selección donde sobrevivirán aquéllas que logren adaptarse a las condiciones desde el sistema in vivo al sistema in vitro. La mayoría de los cultivos celulares de vertebrados no progresan más allá de un número finito de divisiones y entran en senescencia replicativa. En muchas ocasiones, ya sea por inducción o espontáneamente, algunos cultivos celulares pueden adquirir una serie de características proliferativas anormales (independencia de anclaje, pérdida de inhibición de contacto, disminución de la limitación de la densidad de crecimiento, entre otras) proceso denominado transformación celular el cual a menudo se puede correlacionar con la tumorigenicidad. El objetivo general del presente plan de investigación es enunciar nuevos conocimientos en el campo de la Biología Celular y Molecular vinculados con la transformación celular in vitro acontecida en cultivos celulares provenientes de células fetales de conejo (Oryctolagus cuniculus). El cultivo inicial se originó a partir de células obtenidas de piel de la especie en cuestión mediante disgregación enzimática. La línea celular originada a partir de este cultivo se denominó JPCS1. Todos los estudios presentados en esta Tesis se realizaron cada cinco SCs (subcultivos), entre el SC 5 al SC 25 y cada diez SCs desde el SC 30 al SC 50. Para evidenciar la inestabilidad genética los estudios se realizaron tanto a nivel citogenético como molecular. A nivel citogenético se analizaron los cambios en la frecuencia del número cromosómico como así también la evolución del cariotipo de la línea celular durante su mantenimiento entre el SC 5 al 50. A nivel molecular, se utilizó como marcador de inestabilidad genética un conjunto de loci de microsatélites. Las características del control de crecimiento de la línea JPCS1 fueron evaluadas mediante el análisis de las curvas de crecimiento. En las mismas se obtuvo tanto el tiempo de duplicación poblacional media como la densidad limitante de proliferación celular. Asimismo, se realizó el estudio de curvas de crecimiento con independencia de suero donde se obtuvo la pendiente de la curva de crecimiento y la capacidad de proliferación celular en ausencia de suero. Asimismo, también fue analizado el crecimiento independiente de anclaje. Por último, la capacidad de migración celular fue investigada mediante la técnica de la herida. Los resultados de nuestras investigaciones llevados a cabo para dar cumplimiento con los objetivos de la presente Tesis Doctoral demuestran que la misma presenta inestabilidad cromosómica a partir del SC 10, previo a los cambios en los parámetros de crecimiento y migración celular, evidenciando una tendencia al aumento del número cromosómico (hiperdiploidización) cambiando el número modal desde 44 a 46 cromosomas. A partir de ganancias cromosómicas, en primera instancia por una trisomía en el cromosoma 18 a partir del SC 10 y en segunda instancia por trisomía en el cromosoma 6 a partir del SC 20 y posteriormente por tetrasomía del mismo cromosoma en el SC 40. Asimismo, la línea celular muestra un incremento en la frecuencia de la deleción de 10p -del(10)(p13)- a partir del SC 30. A nivel molecular, los resultados obtenidos demuestran que la línea JPCS1 exhibe estabilidad genética del panel de microsatélites utilizados, sin encontrarse nuevos alelos para estos marcadores ni desbalance de las variantes alélicas en los loci heterocigotas. En cuanto a los parámetros de crecimiento celular evaluados, la línea celular en cuestión presenta un acortamiento del ciclo celular demostrando un cambio abrupto entre los SCs 30 y 40, determinado por la disminución del tiempo de duplicación poblacional. Asimismo, se pudo evidenciar la existencia de una tendencia al aumento progresivo de la densidad limitante de la proliferación celular a medida que se incrementa el tiempo de cultivo, exhibiendo una disminución de la inhibición por contacto. Por otra parte, en las etapas más avanzadas del mantenimiento esta línea JPCS1 se caracteriza por exhibir crecimiento con independencia de las necesidades de factores de crecimiento, ya sea por producción autócrina o por presentar algún mecanismo de señalización intracelular alterado. La línea celular JPCS1 no demostró crecimiento con independencia de anclaje en ninguno de los SCs analizados. Por último la misma exhibió un aumento de la capacidad migratoria de las células a partir del SC 25, tendencia que se acentúa en el SC 40 demostrando una disminución de la inhibición por contacto en concordancia con los ensayos de proliferación celular. A partir de los resultados obtenidos en las investigaciones llevadas a cabo durante el desarrollo del presente trabajo de Tesis Doctoral se ha puesto en evidencia, mediante los diferentes estudios realizados que la línea celular JPCS1 originada a partir de células de embriones de Oryctolagus cuniculus luego de su mantenimiento en el sistema in vitro que el cultivo celular se encuentra transformado en el SC 40 proceso que no ha terminado, y que la inestabilidad genética a nivel cromosómico es un evento inicial del proceso de transformación.