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Related Experiment Videos

Localization of a mouse centromeric DNA repeat in interphase nuclei.

C M Disteche1, D A Adler

  • 1Department of Pathology, University of Washington, Seattle 98195.

Cytometry
|January 1, 1990
PubMed
Summary

This study maps mouse DNA repeat locations near centromeres using in situ hybridization. The findings reveal distinct hybridization patterns in male and female mice, varying with X-linked and autosomal loci.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Cell Biology

Background:

  • Centromeric DNA repeats play crucial roles in chromosome structure and function.
  • Understanding the spatial organization of these repeats within interphase nuclei is essential for comprehending genome architecture.

Purpose of the Study:

  • To investigate the interphase nuclear positioning of a specific mouse DNA repeat sequence (70-38) near centromeres.
  • To determine if the location of this repeat varies between X-linked and autosomal loci and across different mouse strains.

Main Methods:

  • In situ hybridization using 3H- or biotin-labeled DNA probe 70-38 on interphase nuclei from mouse bone marrow and fibroblast cells.
  • Analysis of hybridization patterns in different cell cycle phases (G1, early S, late S, G2) using flow-sorted nuclei.

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  • Examination of strains with X-linked locus only versus those with both X-linked and autosomal loci.

    • Main Results:

    • The DNA repeat probe recognized a single locus (DXWas70) near the X chromosome centromere in most strains.
    • Some strains exhibited a second locus on autosomes (D11Was70, D13Was70, or D17Was70).
    • Female mice with only the X-linked locus showed two hybridization sites, while males had one. These sites remained distinct throughout the cell cycle.
    • Mice with both X-linked and autosomal loci displayed four hybridization sites in females and three in males.
    • The DXWas70 and D17Was70 loci were frequently found in the nuclear periphery.

    Conclusions:

    • The spatial organization of centromeric DNA repeats in mouse interphase nuclei is influenced by their chromosomal location (X-linked vs. autosomal).
    • The number and distribution of hybridization sites correlate with sex and the presence of autosomal loci.
    • Specific DNA repeat loci tend to localize to the outer regions of the nucleus.