Direct Detection of Repetitive, Whole Chromosome Paint and Telomere DNA Probes by Immunogold Electron Microscopy

Overview

Journal Chromosome Res

Date 1993 May 1

PMID 8143087

Citations 6

Authors

J Steinmuller

E SCHLEIERMACHER

H Scherthan

Affiliations

Soon will be listed here.

Abstract

Biotinylated repetitive, whole chromosome paint and telomere DNA probes were investigated at the electron microscope level after non-isotopic in situ hybridization and direct immunogold detection. The protocol described allowed the visualization of a biotinylated chromosome 1 specific satellite DNA probe in the light microscope without silver intensification. This sensitive method was exploited to analyse factors contributing to signal strength in immunogold chromosome painting. Furthermore, it allowed us to investigate the distribution of (TTAGGG)n telomere repeats in human metaphase chromosomes and interphase nuclei. Telomeric and internal (TTAGGG)n repeats were detected at high spatial resolution in human metaphase chromosomes. In the periphery of lymphocyte interphase nuclei, two types of telomere hybridization signals were observed. They differed remarkably in compactness, indicating two types of chromatin conformation present at the interphase telomeres in situ.

Citing Articles

Dynamic relocation of telomere complexes in mouse meiotic chromosomes.

Viera A, Parra M, Page J, Santos J, Rufas J, Suja J Chromosome Res. 2004; 11(8):797-807.

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Telomere attachment, meiotic chromosome condensation, pairing, and bouquet stage duration are modified in spermatocytes lacking axial elements.

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An anchorage nuclear structure for telomeric DNA repeats in HeLa cells.

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Yeast nuclei display prominent centromere clustering that is reduced in nondividing cells and in meiotic prophase.

Jin Q, Trelles-Sticken E, Scherthan H, Loidl J J Cell Biol. 1998; 141(1):21-9.

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The telochore: a telomeric differentiation of the chromosome axis.

Suja J, Rufas J Chromosome Res. 1994; 2(5):361-8.

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References

Landegent J, Jansen in de Wal N, Dirks R, Baao F, VAN DER PLOEG M . Use of whole cosmid cloned genomic sequences for chromosomal localization by non-radioactive in situ hybridization. Hum Genet. 1987; 77(4):366-70. DOI: 10.1007/BF00291428. View

Scherthan H, Kohler M, Vogt P, von Malsch K, Schweizer D . Chromosomal in situ hybridization with double-labeled DNA: signal amplification at the probe level. Cytogenet Cell Genet. 1992; 60(1):4-7. DOI: 10.1159/000133282. View

Hutchison N, Ward D, Hamkalo B . In situ hybridization at the electron microscope level: hybrid detection by autoradiography and colloidal gold. J Cell Biol. 1982; 95(2 Pt 1):609-18. PMC: 2112979. DOI: 10.1083/jcb.95.2.609. View

Wieacker P, Davies K, Cooke H, Pearson P, Williamson R, Bhattacharya S . Toward a complete linkage map of the human X chromosome: regional assignment of 16 cloned single-copy DNA sequences employing a panel of somatic cell hybrids. Am J Hum Genet. 1984; 36(2):265-76. PMC: 1684417. View

Sundquist W, Klug A . Telomeric DNA dimerizes by formation of guanine tetrads between hairpin loops. Nature. 1989; 342(6251):825-9. DOI: 10.1038/342825a0. View

Narayanswami S, Hamkalo B . DNA sequence mapping using electron microscopy. Genet Anal Tech Appl. 1991; 8(1):14-23. DOI: 10.1016/1050-3862(91)90004-b. View

Lichter P, Cremer T, Borden J, Manuelidis L, Ward D . Delineation of individual human chromosomes in metaphase and interphase cells by in situ suppression hybridization using recombinant DNA libraries. Hum Genet. 1988; 80(3):224-34. DOI: 10.1007/BF01790090. View

Pluta A, Kaine B, Spear B . The terminal organization of macronuclear DNA in Oxytricha fallax. Nucleic Acids Res. 1982; 10(24):8145-54. PMC: 327075. DOI: 10.1093/nar/10.24.8145. View

Klobutcher L, Swanton M, Donini P, Prescott D . All gene-sized DNA molecules in four species of hypotrichs have the same terminal sequence and an unusual 3' terminus. Proc Natl Acad Sci U S A. 1981; 78(5):3015-9. PMC: 319490. DOI: 10.1073/pnas.78.5.3015. View

10.

Kievits T, Devilee P, Wiegant J, Wapenaar M, Cornelisse C, van Ommen G . Direct nonradioactive in situ hybridization of somatic cell hybrid DNA to human lymphocyte chromosomes. Cytometry. 1990; 11(1):105-9. DOI: 10.1002/cyto.990110112. View

11.

Cooke H, Hindley J . Cloning of human satellite III DNA: different components are on different chromosomes. Nucleic Acids Res. 1979; 6(10):3177-97. PMC: 327928. DOI: 10.1093/nar/6.10.3177. View

12.

de Lange T . Human telomeres are attached to the nuclear matrix. EMBO J. 1992; 11(2):717-24. PMC: 556504. DOI: 10.1002/j.1460-2075.1992.tb05104.x. View

13.

MESSIER P, Jean P, RICHER C . Easy transfer of selected mitoses from light to electron microscopy. Cytogenet Cell Genet. 1986; 43(3-4):207-10. DOI: 10.1159/000132322. View

14.

Williamson J, Raghuraman M, Cech T . Monovalent cation-induced structure of telomeric DNA: the G-quartet model. Cell. 1989; 59(5):871-80. DOI: 10.1016/0092-8674(89)90610-7. View

15.

Henderson E, Hardin C, Walk S, Tinoco Jr I, Blackburn E . Telomeric DNA oligonucleotides form novel intramolecular structures containing guanine-guanine base pairs. Cell. 1987; 51(6):899-908. DOI: 10.1016/0092-8674(87)90577-0. View

16.

Wells R, Germino G, Krishna S, Buckle V, Reeders S . Telomere-related sequences at interstitial sites in the human genome. Genomics. 1990; 8(4):699-704. DOI: 10.1016/0888-7543(90)90257-u. View

17.

Dauwerse J, Wiegant J, Raap A, Breuning M, van Ommen G . Multiple colors by fluorescence in situ hybridization using ratio-labelled DNA probes create a molecular karyotype. Hum Mol Genet. 1992; 1(8):593-8. DOI: 10.1093/hmg/1.8.593. View

18.

Gosden J, Hanratty D, Starling J, Fantes J, Mitchell A, Porteous D . Oligonucleotide-primed in situ DNA synthesis (PRINS): a method for chromosome mapping, banding, and investigation of sequence organization. Cytogenet Cell Genet. 1991; 57(2-3):100-4. DOI: 10.1159/000133122. View

19.

Scherthan H . Localization of the repetitive telomeric sequence (TTAGGG)n in two muntjac species and implications for their karyotypic evolution. Cytogenet Cell Genet. 1990; 53(2-3):115-7. DOI: 10.1159/000132908. View

20.

Collins C, Kuo W, Segraves R, Fuscoe J, Pinkel D, Gray J . Construction and characterization of plasmid libraries enriched in sequences from single human chromosomes. Genomics. 1991; 11(4):997-1006. DOI: 10.1016/0888-7543(91)90025-a. View