Open Access
Rate of Incorporation of R3hlthymidine In Various Tissues of the Mouse A basis for the evaluation of genotoxic effects of chemicals
Author(s) -
Hellman Björn,
Ullberg Sven
Publication year - 1986
Publication title -
cell proliferation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.647
H-Index - 74
eISSN - 1365-2184
pISSN - 0960-7722
DOI - 10.1111/j.1365-2184.1986.tb00729.x
Subject(s) - trichloroacetic acid , thymidine , dna , liquid scintillation counting , chemistry , in vivo , biochemistry , chromatography , dna synthesis , microbiology and biotechnology , biology , genetics
Abstract. [ 3 H]Thymidine has been extensively used as a selective precursor to DNA in studies on the kinetics of cell proliferation. We have become interested in measuring early inhibition of the DNA synthesis in various organs of intact animals for detecting genotoxic properties of chemicals. Such experiments should, for convenience and to achieve a large capacity, be performed in the simplest way possible. The present paper deals with some practical aspects on the use of [ 3 H]thymidine in vivo. [6‐ 3 H]Thymidine was injected intraperitoneally in mice and the uptake of radioactivity was evaluated by using whole‐body autoradiography and liquid scintillation spectrometry. Autoradiograms of sections washed with trichloroacetic acid and methanol Were compared with those subjected only to freeze‐drying. Liquid scintillation counting was performed of total, non‐volatile, acid‐insoluble and dNA‐associated radioactivities. A rapid increase of the [ 3 H]thymidine incorporation was seen during the first hour after the injection. Further prolongation of the survival time did not result in any significant increase of the incorporated radioactivity. Moreover, there were only slight differences between the autoradiograms from extracted and non‐extracted sections. Radioactivities asśociated with DNA closely eorrelated to those representing acid‐insoluble material, indicating that acid‐insoluble radioactivity provides a good estimate of the [ 3 H]thymidine incorporation into DNA.