Hormonal inducibility of liver-specific enzymes in cultured rat embryos.
In monolayer cultures, hepatocyte-specific enzymes are inducible by hormones as soon as hepatocytes differentiate from the embryonic foregut (15-somite stage). Though offering an excellent opportunity for quantitative studies, several features of a normal cell environment are lost in such a model system. To determine the inducibility of such tissue-specific enzymes in intact organisms, rat embryos were cultured in vitro for 48 h and exposed to the hormonal factors that had been found effective in monolayer culture, viz. dexamethasone, triiodothyronine and dibutyryl cyclic AMP. Normal development of the embryos during culture in vitro was assessed by general criteria reflecting growth, morphogenesis and cytodifferentiation. Development of external features, organogenesis, the distribution of cell divisions and the appearance of tissue-specific proteins such as alpha-fetoprotein and glutamate dehydrogenase served as parameters. Despite undisturbed development of the embryos as judged by these criteria, irrespective of whether the culture was started at day 10 or at day 11 of gestation (just before, respectively after the appearance of the liver primordium), induction of hepatocyte-specific enzymes like carbamoylphosphate synthetase by hormones could not be demonstrated immunohistochemically. However, induction of this enzyme by hormones could be demonstrated in monolayers of hepatocytes isolated from such embryos after 48 h of culture, providing yet another demonstration of the adequate culture conditions. In addition, an adequate uptake of hormones by the embryo during culture could be shown with radio-actively labeled dexamethasone and triiodothyronine and with a radioreceptor assay for cyclic AMP. Therefore, the presence of factors in young embryos that inhibit tissue-specific enzyme synthesis has to be postulated.
Westenend
P J
PJ
Department of Anatomy and Embryology, University of Amsterdam, The Netherlands.
Dahmen
R
R
Charles
R
R
Lamers
W H
WH
D000818:Animals; D003994:Bucladesine / Q000494:pharmacology*; D002222:Carbamoyl-Phosphate Synthase (Ammonia) / Q000096:biosynthesis*; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology*; D004622:Embryo, Mammalian; D004790:Enzyme Induction / Q000187:drug effects; D007150:Immunohistochemistry; D008025:Ligases / Q000096:biosynthesis*; D008099:Liver / Q000196:embryology / Q000201:enzymology*; D009024:Morphogenesis; D009924:Organ Culture Techniques; D051381:Rats; D011919:Rats, Inbred Strains; D014284:Triiodothyronine / Q000494:pharmacology*
D014284:Triiodothyronine; D003994:Bucladesine; D003907:Dexamethasone; D008025:Ligases; D002222:Carbamoyl-Phosphate Synthase (Ammonia)