Differential Analysis of Active and Inactive Genes in Human Neutrophils by Chromosomal In Situ Hybridization

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Author(s)

    • IZUMI Shin-ichi
    • Department of Histology and Cell Biology, Nagasaki University School of Medicine
    • SHIN Masashi
    • Department of Histology and Cell Biology, Nagasaki University School of Medicine
    • TAKANO Kunio
    • Department of Oral Histology, Nagasaki University School of Dentistry
    • KOJI Takehiko
    • Department of Histology and Cell Biology, Nagasaki University School of Medicine

Abstract

<i>In situ</i> hybridization has been widely used for localization of specific genes in interphase cell nuclei. The sites of signals, however, do not represent the activity of the gene transcription in the nuclei. To differentially localize genes in active and inactive states, the effect of nuclease digestion prior to <i>in situ</i> hybridization using PCR products of DNA fragments from human X-chromosome, as a probe DNA, was examined in female neutrophils. According to the conventional protocol for chromosomal <i>in situ</i> hybridization, we detected two dot signals in the nucleus. However, the use of Ca/Mg-dependent endonuclease, which extracts inactive gene DNA, resulted in the loss of the one signal of the outer region of the nucleus, but not that of the inner one. In contrast, when exogenous DNase I, which extracts active gene DNA, was used in place of Ca/Mg-dependent endonuclease, the inner signal, but not the outer one, was almost lost in neutrophils. Considering that the outer signal represents inactive X-chromosome DNA in the site of female neutrophil nuclei, these results indicate that the use of our modified protocol may enable us to discriminate the signals of active and inactive genes in interphase cells by <i>in situ</i> hybridization.<br>

Journal

  • ACTA HISTOCHEMICA ET CYTOCHEMICA

    ACTA HISTOCHEMICA ET CYTOCHEMICA 36(4), 325-330, 2003-08-01

    JAPAN SOCIETY OF HISTOCHEMISTRY AND CYTOCHEMISTRY

References:  34

Cited by:  2

Codes

  • NII Article ID (NAID)
    110003144286
  • NII NACSIS-CAT ID (NCID)
    AA00508022
  • Text Lang
    ENG
  • Article Type
    Journal Article
  • ISSN
    00445991
  • Data Source
    CJP  CJPref  NII-ELS  IR  J-STAGE  NDL-Digital 
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