Double labeling autoradiography. Cell kinetic studies with /sup 3/H- and /sup 14/C-thymidine
Examples of the multiple applicability of the double labeling method with /sup 3/H- and /sup 14/C-TdR are demonstrated. Double labeling with /sup 3/H- and /sup 14/C-TdR makes it possible to determine the cycle and its phases with high precision by modifying the usual percent labeled mitoses method with a single injection of /sup 3/H-TdR. In addition, data is provided on the variances of the transit times through the cycle phases. For example, in the case of the jejunal crypt cells of the mouse, the transit times through successive cycle phases are uncorrelated. In the case of glial cells the double labeling method provides cell kinetic parameters despite the paucity of proliferating glial cells. In the adult untreated animal, glial cell mitoses are so rare that the percent labeled mitoses method can not be utilized. However, the S-phase duration can be measured by double labeling and the cycle time can be determined by the so-called method of labeled S phases. With the latter method the passage through the S phase of the /sup 3/H-TdR-labeled S phase cells can be registered by injecting /sup 14/C-TdR at different time intervals following /sup 3/H-TdR application. In this way an S-phase duration of about 10 hr and a cycle time of about 20 hr was found for glial cells in the adult untreated mouse. An exchange of glial cells between the growth fraction and the nongrowth fraction has also been shown by double labeling. A quite different application of the double labeling method with 3H- and /sup 14/C-TdR is the in vivo study of the cell cycle phase-specific effect of drugs used in chemotherapy of tumors. The effect of vincristine on these cells has been studied. Vincristine affects cells in S and G2 in such a manner that they are arrested during the next metaphase and subsequently become necrotic. It has no effect on G1 cells.
- Research Organization:
- Institut fuer Medizinische Strahlenkunde, University Wuerzburg, West Germany
- OSTI ID:
- 5547981
- Journal Information:
- J. Histochem. Cytochem.; (United States), Vol. 29:1A
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
CELL CYCLE
KINETICS
CRYPT CELLS
DOUBLE LABELLING
TUMOR CELLS
AUTORADIOGRAPHY
CARBON 14 COMPOUNDS
MITOTIC INDEX
THYMIDINE
TRITIUM COMPOUNDS
ANIMAL CELLS
AZINES
HETEROCYCLIC COMPOUNDS
LABELLED COMPOUNDS
LABELLING
NUCLEOSIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PYRIMIDINES
RIBOSIDES
SOMATIC CELLS
550301* - Cytology- Tracer Techniques