Precision-cut tissue slices from transgenic mice as an in vitro toxicology system.
In these experiments precision-cut tissue slices from two existing transgenic mouse strains, with transgenes that couple promoting or binding elements to a reporter protein, were used for determination of reporter induction. This approach combines the power of transgenic animals with the practicality of in vitro systems to investigate the biological impact of xenobiotics. Additionally, the normal cellular architecture and heterogeneity is retained in precision-cut tissue slices. Two transgenic mouse strains, one of which couples the promoting region of CYP 1A1 to beta-galactosidase, and another which couples two forward and two backward 12-O-tetradecanoyl phorbol-13-acetate (TPA) repeat elements (TRE) to luciferase (termed AP-1/luciferase), were used to determine the feasibility of this approach. Precision-cut kidney and liver slices from both transgenic strains remain viable as determined by slice K(+) ion content and LDH enzyme release. Liver slices harvested from the CYP 1A1/beta-galactosidase transgenic mice exhibit a 14-fold increase in beta-galactosidase activity when incubated with beta-napthoflavone for 24 h. Kidney and liver slices obtained from the AP-1/luciferase transgenic mice demonstrate induction of luciferase (up to 2.5-fold) when incubated with phorbol myristate acetate (PMA or TPA) up to 4 h. These data indicate that precision-cut tissue slices from transgenic mice offer a novel in vitro method for toxicity evaluation while maintaining normal cell heterogeneity.
Catania
J M
JM
Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, 1703 E. Mabel St., Tucson, AZ 85721-0207, USA.
Parrish
A R
AR
Kirkpatrick
D S
DS
Chitkara
M
M
Bowden
G T
GT
Henderson
C J
CJ
Wolf
C R
CR
Clark
A J
AJ
Brendel
K
K
Fisher
R L
RL
Gandolfi
A J
AJ
D000818:Animals; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics; D004790:Enzyme Induction / Q000187:drug effects; D066298:In Vitro Techniques; D007668:Kidney / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D008156:Luciferases / Q000096:biosynthesis; D051379:Mice; D008822:Mice, Transgenic; D008867:Microtomy; D011401:Promoter Regions, Genetic / Q000235:genetics; D018675:Toxicity Tests / Q000379:methods*; D018808:Transcription Factor AP-1 / Q000235:genetics; D001616:beta-Galactosidase / Q000096:biosynthesis
D018808:Transcription Factor AP-1; D008156:Luciferases; D019363:Cytochrome P-450 CYP1A1; D001616:beta-Galactosidase