Gastrulation is a critical step in vertebrate development, that depends on synergistic effects of several signalling molecules, including fibroblast growth factor-2 (FGF-2). To follow this phenomenon in vitro we isolated rabbit inner cell masses (ICMs) at embryonic day 4 and we exposed ICM-derived cells to FGF-2. Then, we analysed the quantitative differences in rates of protein synthesis from day 3 to day 5 of culture by two-dimensional (2D) gel electrophoresis. Here we show that both up- and down-regulation of protein synthesis took place in ICM-derived cells upon their exposure to FGF-2. The effect of FGF-2 was most pronounced at day 4 of culture, when the changes were very much in favour of a set of down-regulated proteins. To test the significance of this period of time for FGF-2-mediated regulation of protein synthesis, cells were grown without FGF-2 and then they were pulse-treated with FGF-2 at the end of day 4. When compared to the continuous culture with FGF-2, the FGF-2 pulse resulted in a quite indistinguishable pattern of up- and down-regulated proteins. Thus, the readiness of ICM-derived cells to accept and respond to the FGF-2 signals may be of limited duration.

Laboratory of Molecular Embryology Mendel University Brno Czech Republic

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