title: Combined fluorescent-chromogenic in situ hybridization for identification and laser microdissection of interphase chromosomes.
abstract: Chromosome territories constitute the most conspicuous feature of nuclear architecture, and they exhibit non-random distribution patterns in the interphase nucleus. We observed that in cell nuclei from humans with Down Syndrome two chromosomes 21 frequently localize proximal to one another and distant from the third chromosome. To systematically investigate whether the proximally positioned chromosomes were always the same in all cells, we developed an approach consisting of sequential FISH and CISH combined with laser-microdissection of chromosomes from the interphase nucleus and followed by subsequent chromosome identification by microsatellite allele genotyping. This approach identified proximally positioned chromosomes from cultured cells, and the analysis showed that the identity of the chromosomes proximally positioned varies. However, the data suggest that there may be a tendency of the same chromosomes to be positioned close to each other in the interphase nucleus of trisomic cells. The protocol described here represents a powerful new method for genome analysis.
authors.affiliation: Department of Genetics, Faculty of Medicine and Dentistry, University of the Basque Country, Leioa, Spain.
authors.forename: Nerea
authors.initials: N
authors.lastname: Paz
authors.forename: Amaia
authors.initials: A
authors.lastname: Zabala
authors.forename: Félix
authors.initials: F
authors.lastname: Royo
authors.forename: África
authors.initials: Á
authors.lastname: García-Orad
authors.forename: José L
authors.initials: JL
authors.lastname: Zugaza
authors.forename: Luis A
authors.initials: LA
authors.lastname: Parada
mesh_terms: D000483:Alleles; D002460:Cell Line; D002874:Chromosome Mapping / Q000379:methods; D045584:Chromosome Positioning / Q000235:genetics; D002891:Chromosomes, Human, Pair 21; D004314:Down Syndrome / Q000235:genetics; D005838:Genotype; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D007399:Interphase / Q000235:genetics*; D042282:Microdissection / Q000379:methods; D018895:Microsatellite Repeats; D021141:Nucleic Acid Amplification Techniques
references.citation: Lab Invest. 2005 Apr;85(4):582-92
references.pmid: 15696183
references.citation: Genome Biol. 2004;5(7):R44
references.pmid: 15239829
references.citation: Nature. 2007 Jan 25;445(7126):379-781
references.pmid: 17251970
references.citation: Chromosome Res. 2004;12(2):143-51
references.pmid: 15053484
references.citation: J Pathol. 2006 Sep;210(1):3-9
references.pmid: 16823892
references.citation: Cold Spring Harb Perspect Biol. 2010 Mar;2(3):a003889
references.pmid: 20300217
references.citation: Proc Natl Acad Sci U S A. 2007 Sep 18;104(38):14970-5
references.pmid: 17848516
references.citation: Nature. 2007 May 24;447(7143):413-7
references.pmid: 17522674
references.citation: Chromosoma. 2009 Dec;118(6):711-22
references.pmid: 19649645
references.citation: J Cell Biol. 1999 Jun 14;145(6):1119-31
references.pmid: 10366586
references.citation: Chromosoma. 1998 Nov;107(5):330-8
references.pmid: 9880766
references.citation: Am J Pathol. 2000 Nov;157(5):1467-72
references.pmid: 11073807
references.citation: Eur J Hum Genet. 2004 Nov;12(11):907-15
references.pmid: 15292918
references.citation: Semin Cell Dev Biol. 2007 Oct;18(5):707-14
references.pmid: 17905616
references.citation: Nat Rev Mol Cell Biol. 2010 May;11(5):317-28
references.pmid: 20414256
references.citation: Exp Cell Res. 2004 May 15;296(1):64-70
references.pmid: 15120995
references.citation: Science. 1996 May 3;272(5262):725-8
references.pmid: 8614834
references.citation: Expert Rev Mol Diagn. 2008 Jul;8(4):355-7
references.pmid: 18598216
references.citation: Curr Genomics. 2007 Mar;8(1):67-72
references.pmid: 18645627
references.citation: PLoS Biol. 2005 May;3(5):e157
references.pmid: 15839726
references.citation: Am J Hum Genet. 2007 Aug;81(2):252-63
references.pmid: 17668376
references.citation: Nat Methods. 2010 Apr;7(4):299-301
references.pmid: 20305652
references.citation: Biotechniques. 2008 Oct;45(4):385-6, 388, 390 passim
references.pmid: 18855767
references.citation: Biotechniques. 1999 Aug;27(2):362-7
references.pmid: 10457845