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Journal of Lipid Research, Vol 36, 823-838, Copyright © 1995 by Lipid Research, Inc.
ME Pape, PA Schultz, TJ Rea, RB DeMattos, K Kieft, CL Bisgaier, RS Newton and BR Krause
A human cDNA clone (K1) was recently isolated that encodes functional
acyl-CoA:cholesterol acyltransferase (ACAT) protein (Chang et al. J. Biol.
Chem. 1993. 268: 20747-20755). We used the K1 clone to screen a rabbit
liver cDNA library and isolated a 919 base pair partial rabbit cDNA
(ACAT14b) that was greater than 90% homologous with the human nucleotide
sequence. Northern blotting using the rabbit ACAT cDNA14b revealed the
existence of at least six related mRNA species (ranging from 6.2 to 1.7 kb)
in various rabbit tissues. Using an RNAse protection assay, ACAT mRNA14b
was detected in twelve separate rabbit organs. Adrenal gland contained the
highest concentrations of ACAT mRNA14b (per microgram of total RNA) being
20-, 30-, and 50-fold higher than small intestine, aorta, and liver,
respectively. Additional studies with isolated liver cell populations
revealed that rabbit hepatic nonparenchymal cells contained 30-fold more
ACAT mRNA14b (per microgram of total RNA) than parenchymal cells. To
determine whether ACAT mRNA14b levels are regulated in vivo, rabbits were
fed for 4 weeks a high fat/high cholesterol diet (HFHC; 0.5% cholesterol,
3% coconut oil, 3% peanut oil) at which point they were either kept for an
additional 4 weeks on the HFHC-diet or switched to the HFHC-diet plus
CI-976 (50 mg/kg), a potent and specific ACAT inhibitor; another group of
rabbits was fed a chow diet for the entire 8 weeks. The HFHC-diet caused a
2- and 3-fold increase in hepatic and aortic ACAT mRNA14b levels,
respectively, in comparison to chow-fed animals; there was no change in
adrenal or small intestine levels. CI-976 treatment lowered ACAT mRNA14b
levels by 60% and 40% in liver and aorta, respectively, in comparison to
the HFHC controls; again there was no change in adrenal or small intestine
levels. These data indicate that ACAT mRNA14b levels increase in a tissue
specific manner in response to dietary fat and cholesterol.
ARTICLES
Tissue specific changes in acyl-CoA: cholesterol acyltransferase (ACAT) mRNA levels in rabbits
Division of Therapeutics, Parke-Davis Pharmaceutical Research Division, Ann Arbor, MI 48105, USA.
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