Establishment and partial characterization of SV40 virus-immortalized hepatocyte lines of normal and lethal mutant mice carrying a deletion on chromosome 7.
Deletions in chromosome 7 of the mouse have been shown to cause failure of expression of various hepatocyte-specific genes in newborn deletion homozygotes, including the gene encoding tyrosine amino transferase (TAT) (EC 2.6.1.5) (Gluecksohn-Waelsch, 1979). Primary liver cultures of newborn albino deletion mutant mice (c14CoS/c14CoS) and of phenotypically normal mice (c14CoS/cch or cch/cch) were infected with SV40 virus and multiplying hepatocytes selected in arginine-deficient medium containing epidermal growth factor (EGF), insulin, and hydrocortisone (HC). Resulting normal (NMH-ch) and mutant (NMH-m14) hepatocyte lines expressing integrated viral transforming sequences did not senesce, they multiplied autonomously of EGF in medium with insulin plus HC, and they retained hepatocyte-specific functions. Both lines synthesized arginine and contained albumin and alpha-fetoprotein (AFP) mRNAs. TAT-specific mRNA was detected in normal but not in mutant hepatocyte lines. A fragment of the mouse tyrosinase gene, known to map at the albino locus (c) within the region deleted in the c14CoS mutant, hybridized with a 2.5 kb EcoRI fragment of normal NMH-ch DNA, whereas this fragment was undetectable in mutant NMH-m14 DNA. These immortalized hepatocyte lines reflect important properties of normal and mutant liver tissues from which they were derived. The deletion mutant mouse cell lines may be useful for complementation studies involving sequences corresponding to the deletions that encode regulatory gene(s) involved in the control of inducible expression of certain hepatocyte-specific genes such as TAT.
Paul
D
D
Department of Cell Biology, Fraunhofer-Institute for Toxicology and Aerosol Research, Hannover, Federal Republic of Germany.
Kwon
B S
BS
Höhne
M
M
Tönjes
R
R
Haq
A K
AK
Hoffmann
B
B
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D002455:Cell Division; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D002872:Chromosome Deletion; D002874:Chromosome Mapping; D006224:Cricetinae; D015180:Electrophoresis, Gel, Two-Dimensional; D005804:Genes, Lethal; D008099:Liver / Q000166:cytology*; D051379:Mice; D008817:Mice, Mutant Strains; D014176:Protein Biosynthesis; D011506:Proteins / Q000302:isolation & purification; D012313:RNA / Q000235:genetics; D013539:Simian virus 40 / Q000235:genetics
D011506:Proteins; D012313:RNA