engleski

Impact of the Heat Shock on Rat Embryos assessed by Fourier Transform Infrared Spectroscopy

The aim of this study was to explore the possibility of using Fourier Transform Infrared spectroscopy (FTIR) to assess the impact of embryotoxic agents. Different teratogenic agents, among the others elevated body temperature, can severely disturb normal development of the mammalian embryo. At the period of gastrulation this can result in severe congenital malformations of the newborn. This period is therefore recognized as the most critical period of mammalian development. 9, 5-day-old Fisher rat embryos were isolated, cut at the level of amnion and extraembryonic parts were discarded. Embryos were placed in MEM with 50% rat serum, transferred to an CO2 incubator on 42 0C for 15 minutes and after that left at 37 0C. Controls were kept in MEM with 50% rat serum at 37 0C all the time. One group of embryos was immediately processed for FTIR while the second spent 4 hours in serum-supplemented medium on 37 0C and the third group was left for two days in the same medium after being cultivated in disposable tissue culture dish on a metal grid. Before obtaining spectra the serum was washed out and embryos were deposited on silica wafers in a drop of distilled water and dried in vacuum. FTIR spectra were recorded using Perkin-Elmer GX spectrometer supplied with DTGS detector. The changes in spectra of the control group and the heat shocked group were followed in the region of 1800 cm-1 to 600 cm-1 which is assigned to the infrared-active vibrations of nucleic acids (DNA and RNA), proteins and energy rich molecules. Preliminary results suggest that the FTIR spectroscopy may serve as quick and rather easy method for establishing impact of the heat shock on a complex biological system.

Infrared spectroscopy; Heat shock

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