Organization of chromosomes in the mouse nucleus: analysis by in situ hybridization.
We used in situ hybridization with various biotinylated DNA probes to characterize the distribution of mouse chromosomes in the nucleus. This entailed an examination of plasmid-marked chromosomes from two strains of transgenic mice and also an examination of the distribution of centromeres and total mouse genomic DNA. Our results show that, regardless of the hybridization probes used or the tissue types examined, with sections of mouse tissues embedded in paraffin all chromosomal DNA appeared to be localized to the nuclear periphery. This was observed in tissues fixed with either crosslinking or non-crosslinking fixatives, and submitted to a variety of prehybridization treatments. Further experiments with non-embedded nuclei fractionated from mouse liver revealed a similar distribution of DNA at the telomeric end of chromosome 3, but for centromeric DNA and total mouse genomic DNA, hybridization signals were observed both in the interior and at the periphery of the nucleus. These observations preclude the exclusive localization of chromosomes to the nuclear periphery. However, they indicate that a subset of centromeres are likely to be associated with the nuclear membrane, and that there is such an association at the telomeric end of chromosome 3. Overall, these results are compatible with previous observations of other investigators showing regions of contact between chromosomes and the nuclear membrane.
Katsumata
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Biology Department/G5, University of Pennsylvania, Philadelphia 19104.
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D000818:Animals; D001921:Brain / Q000648:ultrastructure; D002467:Cell Nucleus / Q000648:ultrastructure*; D002503:Centromere / Q000648:ultrastructure; D002875:Chromosomes / Q000648:ultrastructure*; D004247:DNA / Q000032:analysis; D007421:Intestine, Small / Q000648:ultrastructure; D007668:Kidney / Q000648:ultrastructure; D008099:Liver / Q000648:ultrastructure; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008854:Microscopy, Electron; D009693:Nucleic Acid Hybridization; D013737:Testis / Q000648:ultrastructure
D004247:DNA