and strategies for avoidance. 1, C and D). None of these genes revealed altered expression in c-jun / mice (Fig. Primary antibodies were detected with fluorescein isothiocyanate-conjugated or tetramethylrhodamine isothiocyanate-conjugated antibodies directed against mouse and rat or rabbit immunoglobulins, respectively. Table I Artificial RNA Standards and Primers for Quantitative RT-PCR Morphologic and Immunohistochemical Analysis of Fetuses Mouse fetuses were fixed in 10 phosphate-buffered formaldehyde, paraffin-embedded, and then 4-m sections were stained with hematoxylin-eosin (HE). Male F344/BN rats were assigned to one of four groups. Primary fibroblasts from E12.5 fetuses were cultured as described ( Robertson, 1987 ). These alterations could, in principle, also reflect a delay in heart development rather than a true malformation. Note the thinner wall of the ventricles in c-jun / hearts (H). The data obtained in vitro together with the observation that several c-jun / E13.0 fetuses showed increased apoptoses of erythroblasts and hepatoblasts in their livers, point to an essential role of c-Jun in the regulation of apoptosis in a diversity of cell types in vitro. The amount of c-jun mRNA was estimated by quantitative RT-PCR in wild-type mouse livers of different age (E12.5-adult).
Here we show that a fraction of mutant E13.0 fetal livers exhibits extensive apoptosis of both hematopoietic cells and hepatoblasts, whereas the expression of 15 mRNAs, including those of albumin, keratin 18, hepatocyte nuclear factor 1, -globin, and erythropoietin, some of which are putative AP-1. Abbreviations used in this paper: AP-1 activating protein 1 Cx 43 connexin 43 ES cells embryonic stem cells GPI glucose phosphate isomerase HGF hepatocyte growth factor HNF-1 hepatocyte nuclear factor 1 RT-PCR reverse transcriptase PCR tunel TdT-mediated dUTP nick-end labeling Submitted: Revision received References Eferl. Presence of c-jun / hepatoblasts during fetal development was analyzed in short-term cultures of fetal liver cells and average chimerism was determined in the residual fetus (after removal of the liver).
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The proliferation defect in c-jun / fibroblast cell lines was found to be p53-dependent, indicating that the alterations of proliferation, and probably also the increased propensity of cells to undergo apoptosis may involve p53-dependent pathways. Moreover, as shown by neural crest cell transplantation experiments, the mere presence of neural crest cells does not exclude functional deficiencies ( Kirby and Waldo, 1990 ). Schüle., 1990 and MyoD bengal., 1992 ) yielding a network of transcriptional regulation. The next major step in mouse liver development occurs at approximately E14.5 when hepatoblasts start to differentiate into the hepatocytic and bile duct epithelial lineage, which is indicated by the formation of the ductal plate, which later differentiates into the intrahepatic bile ducts ( Desmet. First strand cDNA synthesis for quantitative RT-PCR was performed in a 20-l reaction mixture containing.1 g of total RNA (isolated as described by Krieg., 1983.5 units Inhibit-ACE (5 3 Inc. This is in agreement with the reduced mitotic and increased apoptotic rates found in primary liver cell cultures derived from c-jun / fetuses. Moreover, bile duct epithelial cells were analyzed after enrichment by microdissection. The growth rates and the achieved cell densities were markedly reduced in c-jun / cultures (Fig. Figure 3 Analysis of mRNA expression in E12.5 wild-type ( c-jun heterozygous ( c-jun and homozygous c-jun deleted ( c-jun fetal livers by quantitative RT-PCR. Ischaemiareperfusion (I-R) was chosen as a stressor because I-R is known to generate reactive oxygen and nitrogen species (rons) and to activate inflammatory cells ; ). Fetal livers were dissected from staged fetuses and after a brief rinse in PBS subjected to 5-min subsequent incubations in solution A (ebss without Ca2 and Mg2 containing.5 mM egta solution B (ebss containing Ca2, Mg2, and 10 mM Hepes,.4 and solution.
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