Gene transfer into Xenopus hepatocytes: transcriptional regulation by members of the nuclear receptor superfamily.
A procedure to culture Xenopus laevis hepatocytes that allows the cells in primary culture to be subjected to gene transfer experiments has been developed. The cultured cells continue to present tissue-specific markers such as expression of the albumin gene or estrogen-controlled vitellogenin gene expression, which are both restricted to liver. Two efficient and reproducible gene transfer procedures have been adapted to the Xenopus hepatocytes, namely lipofection and calcium phosphate-mediated precipitation. The transcription of transfected reporter genes controlled by estrogen-, glucocorticoid- or peroxisome proliferator-response elements was stimulated by endogenous or co-transfected receptor in a ligand-dependent manner. Furthermore, the expression of a reporter gene under the control of the entire promoter of the vitellogenin B1 gene mimicked the expression of the chromosomal vitellogenin gene with respect to basal and estrogen-induced activity. Thus, this culture-transfection system will prove very useful to study the regulation of genes expressed in the liver under the control of various hormones or xenobiotics.
Marilley
D
D
Institut de Biologie Animale, Université de Lausanne, Switzerland.
Mahfoudi
A
A
Wahli
W
W
D000418:Albumins / Q000235:genetics; D000818:Animals; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D008099:Liver / Q000502:physiology*; D008297:Male; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics*; D014158:Transcription, Genetic; D014819:Vitellogenins / Q000235:genetics; D014982:Xenopus laevis / Q000502:physiology*
D000418:Albumins; D018160:Receptors, Cytoplasmic and Nuclear; D014819:Vitellogenins