Changes in some chromatin and cytoplasmic enzymes of perinatal rat hepatocytes during culture |
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Authors: | C Guguen-Guillouzo L Tichonicky M F Szajnert J Kruh |
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Institution: | 1. Unité de Recherches Hépatologiques, INSERM U 49 H?pital Pontchaillou, 35011, Rennes Cédex, France 2. Institut de Pathologie Moléculaire, INSERMU 137, 24, rue du faubourg Saint-Jacques, 75014, Paris, France
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Abstract: | Summary Hepatocytes prepared from rats at various perinatal stages were cultured in selective medium that does not allow fibroblastic
cell growth. Cell population remained homogeneous during the culture. Hepatocytes undergo divisions for a period, which varies
according to the stage of development of the rat. Light and electron microscope observations showed the presence of numerous
cytoplasmic organelles; moreover, hydrocortisone-induced structures similar to bile canaliculi. Chromatin protein kinase decreased
rapidly during culture except in samples prepared from 17-day fetuses in which it remained unchanged for 2 days and decreased
to a lesser extent afterwards. Chromatin nonhistone proteins were incubated with (γ-32P) ATP and the phosphorylation pattern analyzed on polyacrylamide gels. Many radioactive peaks were observed in chromatin
proteins from 17-day fetuses; they were much lower in proteins from 19-day fetuses. The phosphorylation pattern was analyzed
in hepatocytes after 2 days of culture. Many radioactive peaks were observed with proteins from heapatocytes taken from 17-day
fetuses; no radioactivity was observed in proteins from 19-day fetuses. This is in contrast with the absence of radioactive
peaks in chromatin proteins from adult rat hepatocytes.
In cytoplasm, aldolase and pyruvate kinase specific activities varied according to the age of the rat. They strongly decreased
during culture except in hepatocytes from 15-and 17-day fetuses, in which they remained stable for at least 5 days. The stability
of chromatin and cytoplasmic enzymes in hepatocytes from 17-day fetuses could result from their ability to be regulated by
hormones that are secreted at this stage of development. |
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Keywords: | fetus rat hepatocytes primary culture chromatin enzymes |
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