Molecular cytogenetic characterization of the mouse cell line WMP2 by spectral karyotyping and multicolor banding applying murine probes

Abstract

The Moloney murine leukemia virus-transformed suspension cell line WMP2 is derived from wild mice (Mus musculus) of the WMP/WMP strain. These mice carry nine pairs of metacentric Robertsonian translocation chromosomes. As the chromosomes of the wild-type mouse are all acrocentric, metaphase spreads of the WMP2 cells seam to be highly suited for physical gene mapping. Here we studied the WMP2 line using spectral karyotyping (SKY) combined with new established mouse specific multicolor banding (mcb) probes for the chromosomes X, 3, 4, 6 and 18. SKY revealed that the WMP2 cell line developed further four derivative chromosomes. After application of mcb five previously unrecognizable intrachromosomal rearrangements with 9 breakpoints were detected for the studied chromosomes.

New insights into the karyotypic evolution in muroid rodents revealed by multicolor banding applying murine probes

“…For cross-species hybridizations, mcb probe sets for chromosomes 3, 6, 18, and 19 available at the Institute of Human Genetics and Anthropology (Jena), were applied as previously described Trifonov et al 2005, Karst et al 2006. A twofold-increased probe concentration and hybridization for up to 96 h was necessary for some hybridization.…”

“…Recently, we described the generation of similar libraries for murine multicolour banding (mcb-the lower case is used here to distinguish these probes from MCB with human probes; Trifonov et al 2005;Karst et al 2006), as also reported by Benedek et al (2004). Here, we applied four selected autosomal mcb probes to study the evolution of chromosomes 3, 6, 18, and 19 in nine muroid species, belonging to subfamilies Murinae (Norway rat, striped field mouse), Cricetinae (golden, Chinese, greater long-tailed hamsters), and Arvicolinae (tundra vole, southern vole, collared lemming, mole vole).…”

First Molecular Cytogenetic High Resolution Characterization of the NIH 3T3 Cell Line by Murine Multicolor Banding

“…As FISH approaches using murine wcp probes are limited when the exact localization of chromosomal breakpoints is required, a previously reported, more sensitive approach to murine FISH banding, the murine multicolor banding (mcb) (Trifonov et al 2005; Karst et al 2006; Trifonov et al 2010), was established for all murine chromosomes.…”

Generation of multicolor banding probes for chromosomes of different species

“…When the doubles were excluded we got 19 unique wcp probes for autosomes and wcp probes for the X and Y chromosomes (which were identified earlier). To detect which wcp corresponded to which chromosome we have used several different approaches: 1) wcp probes were applied onto metaphases from WMP2 cell line known to contain easily identifiable metacentric fusion chromosomes characterized before by G-banding and FISH [8,13]; 2) wcp probes were co-hybridized with murine BAC probes available in our laboratory 3) wcp probes were co-hybridized with previously obtained murine region specific probes [8]. After all wcp probes were successfully attributed, they were further used in FISH-MD experiments to generate region-specific pcp paints required for mcb probe sets.…”

“…This created the necessity of generating MCB probes for the chromosomes of other species, potentially interesting from the cytogenetic point of view. The first attempt to generate FISH-banding probes for non-human chromosomes was made by our group in 2002, when multicolor banding probes for mouse chromosomes 3, 6, 18, 19 and X were established; published in 2006 [8]. To distinguish murine/non human multicolor banding probes from the previously established MCB probes for human chromosomes, it was suggested to use lowercase letters, as opposed to capital letters, in the abbreviation – “mcb” (the same was suggested for non-human WCP probes – “wcp”).…”