Karyotype analysis

Goldstein, P. and Triantaphyllou, A.C. (1979) Karyotype analysis of the plant-parasitic nematode Heteroderaglycinesby electron microscopy. II. The tetraploid and an aneuploid hybrid. Journal of Cell Science 43, 225. 

During mitosis, aU the DNA is highly condensed to allow separation of the sister chromatids. This is the only time in the ceE cycle when the chromosome structure is visible. Chromosome abnormalities may be assessed on mitotic chromosomes by karyotype analysis (metaphase chromosomes) and by banding techniques (prophase or prometaphase), which identify aneu-ploidy, translocations, deletions, inversions, and duplications. 

Karyotypic analysis Oncogene screening Gene stability Infection DNA screens... 

Mitterbauer M, Kusec R, Schwarzinger I, Haas OA, Lechner K, )aeger U. Comparison of karyotype analysis and RT-PCR for AMLl/ETO in 204 unselected patients with AML. Ann Hematol 1998 76 139-43. 

A particularly relevant study by De Carli et al. (D9) is described. A karyotype analysis was made of both a very high alkaline phosphatase strain of an embryonic clone of strain EUE cells and two sublines (1 and 2) of cells exhibiting very low activity. Cells of lines Sub 1 and Sub 2 show a consistent reduction of one or two units in long acrocentric and a much more pronounced reduction in short acrocentric chromosomes. The high EUE cells accordingly have twice as many short acrocentric chromosomes. Finally, there was no simple relationship between chromosome dose and levels of enzyme activity in these cell lines, although this relationship has been seriously considered by Alter et al. (A12, A13) for the leukocytes. 

The answer is c. (Murray, pp 812-828. Scriver, pp 3-45. Sack, pp 57-76. Wilson, pp 123-149.) Fluorescent in situ hybridization (FISH) analysis is a technique in which molecular probes that are specific for individual chromosomes or chromosomal regions are used to identify these regions. FISH probes frequently identify chromosomal regions that are submicro-scopic and therefore may be useful when standard karyotypic analysis is normal. In this case, the fact that only one signal is present, despite the fact that there are two number 22 chromosomes, indicates that a submicro-scopic deletion has occurred. The parental chromosome of origin cannot be determined using this technique unless that parent also carries a similar deletion and his or her chromosomes are evaluated. 

Kawauchi S, Fukuda T, Miyamoto S, et al. Peripheral primitive neuroectodermal tumor of the ovary confirmed by CD99 immunostaining, karyotypic analysis, and RT-PCR for EWS/FLI-1 chimeric mRNA. Am J Surg Pathol. 1998 22 1417-1422. 

Fig. 2.4 Chromosome analysis of the heterohybri-doma CB03. GTG banding (upper left), spectral karyotype analysis (upper right) and identification of human chromosomes by hybridization with specifically labeled human chromosome libraries...

Strategies to experimentally detect translocations are impoilant because of the numerous cases of genes in leukemia-associated translocations. Such methods include Southeni blot analysis, which is not as sensitive as PCR, karyotype analysis and fluorescence in situ hybridization (FISH) with specific probes. Revei se transcriptase (RT)-PCR with gene-specific primers detects only a fraction of translocations because there are no primers available for many of the genes involved. 

It must be emphasized that the dominant lethal assay should be applied together with other in vivo- mammalian mutagenicity tests, notably in vivo cytogenetics, karyotype analysis of bone marrow, and the host-mediated assay, in addition to ancillary submammalian tests. Such mammalian systems can be simply and practically incorporated in the course of routine toxicity testing. 

Schwarzacher, T., Ambros, P., and Schweizer, D. (1980) Application of Giemsa banding to orchid karyotype analysis. Plant Syst. Evol. 134,293-297. 

PCR for the ZJy gene detects the presence of a Y chromosome. However, it should be noted that an EG cell line that is negative for this PCR product may actually be an XO cell line, not a normal XX female cell line. It would be necessary to perform karyotype analysis to determine the difference between an XO and an XX cell line accurately. 

Skinner D.Z., Budde, A.D. and Leong, S.A. (1991) Molecular karyotype analysis of fungi. In Bennet, J.W. and Lasure, L.L.(eds.) More Gene Manipulation in Fungi. Academic Press, New. York, p. 86-98. 

K. R. Castleman, J. Melnyk, and H. J. Frieden, Karyotype analysis by computer and its application to mutagenicity testing of environmental chemicals, Mutat. Res. 41, 153-161... 

Saylor, L.C. 1972. Karyotype analysis of the genus Finns - subgenus Finns. Silvae Genet. 19 155-163. 

Nkongolo, K.K., and K. Klimaszewska. 1993. Karyotype analysis and optimization of mitotic index in Picea mariana (black spruce) preparations from seedling root tips and embryonic cultures. Heredity 73 11-17. 

Karyotype Cells were treated with 0,1 pg Colcemid in 20 minutes and incubated in KCl 0,075M. Following this, cells were fixed in fixative s solution (methanol acetic acid). G banding technique Slides were treated in 0,5% Trypsin and then in 5% staining Giemsa s solution in 5 minutes. The results were analyzed by Cytovision, Nikon 600 karyotyping analysis system. 

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