Regulation of protein synthesis by estradiol 17β, dexamethasone and insulin in primary cultured Xenopus hepatocytes

Kawahara, Akira; Sato, Kenzo; Amano, Minoru

doi:10.1016/0014-4827(83)90164-7

Cited by 14 publications

(5 citation statements)

References 23 publications

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“…We are not aware of a similar specific and reversible deinduction of expression of other genes. It is worth noting that the deinduction is hormonespecific, since glucocorticoid hormone has been shown to promote the synthesis and secretion of albumin by primary cultures of Xenopus hepatocytes [43,50], although short-term studies similar to those we report here have not been performed with other hormones.…”

Section: Discussionmentioning

confidence: 62%

Deinduction of transcription of Xenopus 74-kDa albumin genes and destabilization of mRNA by estrogen in vivo and in hepatocyte cultures

et al. 1985

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The goal of this study is to explain the molecular basis of the marked deinduction of Xenopus albumin synthesis and secretion accompanying the activation of vitellogenin genes by estrogen. We have characterized by restriction analysis, DNA sequencing and hybrid-selected translation of mRNA, a cloned cDNA specifying the two 74-kDa albumins which constitute the predominant circulating form of albumin in Xenopus luevis. Using this recombinant DNA plasmid as a hybridization probe, we have determined the steady-state levels of albumin mRNA, the rate of transcription of the two 74-kDa albumin genes and the stability of the mRNA in male and female Xenopus hepatocytes in vivo and in primary cell cultures following estrogen treatment.In both whole liver and cultured hepatocytes estradiol caused a rapid drop in the steady-state levels of 74-kDa albumin mRNAs, which was reversed spontaneously in the continued presence of the hormone. The concentration of albumin mRNA was substantially higher in male than in female hepatocytes, the hormonal effect being more marked in male than in female hepatocytes.The decrease in steady-state levels of mRNA was anticipated in male hepatocytes by a 70% inhibition of rate of transcription of albumin genes within 2 h of exposure to estradiol, as measured by run-off transcription in liver nuclei isolated from animals treated in vivo or by determining the absolute transcription rate in cell cultures. In the latter the diminished transcription rate returned to normal within 12 h in the continued presence of the hormone.Estradiol caused a threefold destabilization of albumin mRNA in both male and female hepatocyte cultures to tliz = 3 h and 2 h respectively. The combined effects on rate of transcription and mRNA stability largely explain the changes in the steady-state levels of mRNA caused by hormone administration.Comparison of the kinetics of transcription rates of vitellogenin and albumin genes in vivo and in vitro reveals a striking reciprocity in the selective activation of the inducible genes and deinduction of the constitutively expressed genes at the early stages of response of Xenopus hepatocytes to estrogen.

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Section: Discussionmentioning

confidence: 62%

Deinduction of transcription of Xenopus 74-kDa albumin genes and destabilization of mRNA by estrogen in vivo and in hepatocyte cultures

et al. 1985

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“…Glucocorticoids, which are known to enhance albumin synthesis [81,82], help to maintain its rate when added to hepatocytes soon after their isolation [50,83]. In this respect the full induction of vitellogenin synthesis in primary cultures of amphibian or avian liver parenchymal cells offers many advantages, such as the competence of male liver to respond to estrogen, offering a'zero' background of prior vitellogenin gene expression and being independent of cell division There are many similarities between hormonal induction of gene expression and the induction by drugs of detoxicating enzyme systems [89,90].…”

Section: Hormonal Responsiveness In Short Term Culturesmentioning

confidence: 99%

“…The levels of hormonally regulated tissue specific mRNAs and proteins may rapidly decline following cell isolation, e.g., albumin [46] or LYW globulin [80]. Glucocorticoids, which are known to enhance albumin synthesis [81,82], help to maintain its rate when added to hepatocytes soon after their isolation [50,83]. In this respect the full induction of vitellogenin synthesis in primary cultures of amphibian or avian liver parenchymal cells offers many advantages, such as the competence of male liver to respond to estrogen, offering a'zero' background of prior vitellogenin gene expression and being independent of cell division or DNA synthesis [72,84,85].…”

Section: Hormonal Responsiveness In Short Term Culturesmentioning

confidence: 99%

Primary culture, cellular stress and differentiated function

Wolffe

Tata

1984

FEBS Letters

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Isolation of specialized cell types for the analysis of tissue‐specific gene function often results in loss of the differentiated phenotype. Examples of this type of phenotypic change following tissue disaggregation are reviewed together with possible explanations. Close similarities between the effects of cell isolation with those of other cellular stresses such as heat or anoxia point to common biochemical mechanisms being involved. This suggests that the study of freshly isolated cells will contribute significantly to out understanding of the nature of cellular stress and its consequences for the maintenance of phenotype and induction of tissue specific gene expression.

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“…Hepatocytes were isolated from adult male X. laevis as described by Kawahara et al 16) . A one-step perfusion method was used as follows.…”

Section: Primary Culturementioning

confidence: 99%

“…With regard to the abovementioned point, in vitro primary hepatocyte assay detecting hepatic vitellogenin(VTG)as a biomarker might be useful 15,16) . We have proposed VTG as an ideal biomarker in the screening of estrogenic activities of endocrine disrupters in a wide range of oviparous animals 17) .…”

Section: Introductionmentioning

confidence: 99%

Estrogenic Activity of Phthalate Esters by In Vitro VTG Assay Using Primary-cultured Xenopus Hepatocytes

et al. 2006

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Regulation of protein synthesis by estradiol 17β, dexamethasone and insulin in primary cultured Xenopus hepatocytes

Cited by 14 publications

References 23 publications

Deinduction of transcription of Xenopus 74-kDa albumin genes and destabilization of mRNA by estrogen in vivo and in hepatocyte cultures

Deinduction of transcription of Xenopus 74-kDa albumin genes and destabilization of mRNA by estrogen in vivo and in hepatocyte cultures

Primary culture, cellular stress and differentiated function

Estrogenic Activity of Phthalate Esters by In Vitro VTG Assay Using Primary-cultured Xenopus Hepatocytes

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