title: Chromosome Banding in Amphibia. XXXIII. Demonstration of 5-Methylcytosine-Rich Heterochromatin in Anura.
abstract: An experimental approach using monoclonal anti-5-methylcytosine (5-MeC) antibodies and indirect immunofluorescence was elaborated for detecting 5-MeC-rich chromosome regions in anuran chromosomes. This technique was applied to mitotic metaphases of 6 neotropical frog species belonging to 6 genera and 4 families. The hypermethylation patterns were compared with a variety of banding patterns obtained by conventional banding techniques. The hypermethylated DNA sequences are species-specific and located exclusively in constitutive heterochromatin. They are found in centromeric, pericentromeric, telomeric, and interstitial positions of the chromosomes and adjacent to nucleolus organizer regions. 5-MeC-rich DNA sequences can be embedded both in AT- and GC-rich repetitive DNA. The experimental parameters that have major influence on the reproducibility and quality of the anti-5-MeC antibody labeling are discussed.
authors.affiliation: Department of Human Genetics, University of Wx00FC;rzburg, Wx00FC;rzburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Schmid
authors.forename: Claus
authors.initials: C
authors.lastname: Steinlein
mesh_terms: D044503:5-Methylcytosine / Q000032:analysis* / Q000276:immunology; D020856:AT Rich Sequence / Q000235:genetics; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001001:Anura / Q000145:classification / Q000235:genetics*; D002503:Centromere / Q000235:genetics; D002871:Chromosome Banding / Q000379:methods* / Q000592:standards; D019175:DNA Methylation; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods* / Q000592:standards; D020862:GC Rich Sequence / Q000235:genetics; D006570:Heterochromatin / Q000737:chemistry* / Q000276:immunology; D059785:Karyotype; D008677:Metaphase; D008938:Mitosis; D009697:Nucleolus Organizer Region / Q000235:genetics; D015203:Reproducibility of Results; D013045:Species Specificity; D016615:Telomere / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D006570:Heterochromatin; D044503:5-Methylcytosine