MICROFLUOROMETRIC ANALYSIS OF DEOXYRIBONUCLEIC ACID REPLICATION KINETICS AND SISTER CHROMATID EXCHANGES IN HUMAN CHROMOSOMES

Fluorescence of the dye 33258 Hoechst, when bound to chromosomes, is partially quenched by the incorporation of 5-bromodeoxyuridine into chromosomal deoxyribonucleic acid (DNA). This effect allows microfluorometric analysis of DNA synthesis. Metaphase chromosomes from cultured human leukocytes which...

Full description

Saved in:
Bibliographic Details
Published in:The journal of histochemistry and cytochemistry 1974-07, Vol.22 (7), p.478-491
Main Author: LATT, SAMUEL A.
Format: Article
Language:English
Subjects:
Bromodeoxyuridine - pharmacology
Cell Division
Chromatids - drug effects
Chromatids - metabolism
Chromosomes - drug effects
Chromosomes - metabolism
DNA - biosynthesis
DNA - blood
DNA Replication
Female
Floxuridine - pharmacology
Histocytochemistry
Humans
Karyotyping
Leukocytes - cytology
Leukocytes - drug effects
Leukocytes - metabolism
Microscopy, Fluorescence
Mitosis
Quinacrine
Staining and Labeling
Thymidine - metabolism
Time Factors
Tritium
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Fluorescence of the dye 33258 Hoechst, when bound to chromosomes, is partially quenched by the incorporation of 5-bromodeoxyuridine into chromosomal deoxyribonucleic acid (DNA). This effect allows microfluorometric analysis of DNA synthesis. Metaphase chromosomes from cultured human leukocytes which have incorporated 5-bromodeoxyuridine for a portion of the DNA synthesis period exhibit reduced 33258 Hoechst fluorescence in 5-bromodeoxyuridine-containing regions. Regions synthesizing DNA during a particular interval can thus be highlighted by the appropriate protocol of 5-bromodeoxyuridine administration. Chromosomes from cells which have replicated twice in medium containing 5-bromodeoxyuridine exhibit one brightly and one dully fluorescing chromatid, reflecting incorporation of 5-bromodeoxyuridine into one or two chains of chromatid DNA, respectively. Sister chromatid exchanges, evident as sharply demarcated reciprocal alterations in fluorescence along chromosomes, can be located relative to quinacrine banding patterns. This fluorometric approach should be useful in many instances as a convenient, high resolution alternative to autoradiography.
ISSN:0022-1554
1551-5044
DOI:10.1177/22.7.478