A rapid procedure has been devised to count the numbers of outer trophectoderm (TE) and inner cell mass (ICM) cells of mouse blastocysts by differentially labelling their nuclei in situ with polynucleotide-specific fluorochromes. The TE nuclei were labelled with propidium iodide (PI) by permeabilisi
Difference in sensitivity of inner cell mass and trophectoderm to X-irradiation in mouse blastocysts
โ Scribed by Kumazawa, Toshihiko; Inouye, Minoru; Hayasaka, Ikuo; Yamamura, Hideki; Murata, Yoshiharu
- Publisher
- John Wiley and Sons
- Year
- 1998
- Tongue
- English
- Weight
- 359 KB
- Volume
- 57
- Category
- Article
- ISSN
- 0040-3709
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โฆ Synopsis
Pregnant B6C3F1 mice were exposed to a single whole body X-irradiation on day 4 (73-74 hr postconception) of gestation. In experiment 1, they were sacrificed at 2, 4, 6, or 9 hr after a dose of 2 Gy, and their embryos were removed and examined with light and electron microscopy. In experiment 2, dose-response effects of irradiation on the embryos were examined 4 hr after doses of 0-4 Gy. In experiment 3, DNA fragmentation (a marker of apoptosis) was observed by 3ะ-OH nick-end labeling technique. In inner cell mass (ICM) and trophectoderm (TE) of blastocysts exposed to 2 Gy, cells with cytoplasmic degeneration, or dead cells phagocytosed by their neighboring cells, were found. Although morphological features of these dying cells did not reveal typical characteristics of apoptosis such as nuclear condensation and membrane blebbing, DNA fragmentation was detected by nick-end labeling technique. The degenerated cytoplasm consisted of aggregating ribosomes. Degenerated cells began to increase from 2 hr after irradiation and reached maximal at 4 hr in both ICM and TE. The incidences of degenerated cells in ICM were higher than those in TE at any time point. These findings provide evidence that cell death observed in blastocysts after X-irradiation is apoptotic and sensitivity of the two groups of cells (ICM and TE) to X-rays is different.
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