title: Analysis of Meiotic Telomere Behavior in the Mouse.
abstract: Linear eukaryotic chromosomes are capped by the telomeres, which consist of highly repetitive nucleotide sequences bound by several telomere-specific proteins. While the general role of telomeres is to protect chromosomes from degradation and end-to-end fusion, during meiosis they are assigned with a distinct and without doubt highly fascinating function. During meiosis, telomeres attach to the nuclear envelope and mediate characteristic chromosome movements, essential for correct haploidization of the genome. Here, we provide elaborate tools to study telomeres in mammalian meiotic germ cells, which include (co-)immunofluorescence staining procedures on cell spreads and paraffin-embedded tissues. We provide detailed procedures for fluorescence labeling of telomeric DNA (Telo-FISH) to visualize telomeres at the light microscopic level, which we often use in combination with immunofluorescence staining of meiotic proteins. We also present a protocol for detection of telomeric DNA at the electron microscopic level (EM-ISH). We finally describe how meiotic telomeres can be visualized by common electron microscopic methods and how they can be analyzed at the ultrastructural level by immunogold labeling of telomere components or associated structures.
authors.affiliation: Department of Cell and Developmental Biology, Biocenter, University of Würzburg, Am Hubland, 97074, Würzburg, Germany.|Department of Chromosome Biology, Max F. Perutz Laboratories (MFPL), Dr. Bohr-Gasse 9, 1030 Vienna, Austria.
authors.forename: Jana
authors.initials: J
authors.lastname: Link
authors.affiliation: Department of Cell and Developmental Biology, Biocenter, University of Würzburg, Am Hubland, 97074, Würzburg, Germany.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Benavente
authors.affiliation: Department of Cell and Developmental Biology, Biocenter, University of Würzburg, Am Hubland, 97074, Würzburg, Germany. alsheimer@biozentrum.uni-wuerzburg.de.
authors.forename: Manfred
authors.initials: M
authors.lastname: Alsheimer
mesh_terms: D000818:Animals; D033481:Chromosomes, Mammalian; D005260:Female; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008540:Meiosis / Q000235:genetics*; D051379:Mice; D008854:Microscopy, Electron; D009685:Nuclear Envelope / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism; D010053:Ovary / Q000378:metabolism; D013090:Spermatocytes / Q000378:metabolism; D016615:Telomere / Q000235:genetics*; D059505:Telomere Homeostasis; D013737:Testis / Q000378:metabolism
keywords: EM-ISH; Electron microscopy; FISH; Immunofluorescence analysis; Meiosis; Nuclear envelope; Telomere