Polymorphic debrisoquine 4-hydroxylase activity in the rat is due to differences in CYP2D2 expression.
The female Dark Agouti rat is widely used as an animal model for the CYP2D6 poor metabolizer phenotype, males of other strains such as Sprague Dawley or Wistar serving as models for the extensive metabolizer phenotype. To determine the relative level of expression of CYP2D enzymes in the liver of female and male Dark Agouti, Sprague Dawley and Wistar rats, anti-peptide antibodies were raised in rabbits against short synthetic peptides representing the C-termini of the rat P450 enzymes CYP2D1, CYP2D2, CYP2D3, CYP2D4 and CYP2D5. In immunoblotting studies, it was found that the hepatic expression of CYP2D1 was greater in Dark Agouti rats than Sprague Dawley or Wistar rats. In contrast, hepatic CYP2D2 was 30-40-fold less abundant in female Dark Agouti than female Sprague Dawley or Wistar rats and six- to eightfold less abundant in male Dark Agouti than male Sprague Dawley or Wistar rats. No hepatic CYP2D3 could be detected in either sex of any of the three strains. Hepatic CYP2D4 expression was generally greater in male than female rats, and higher in Dark Agouti compared with Sprague Dawley or Wistar strains. CYP2D5 was expressed in the livers of female and male Dark Agouti rats but not in female Sprague Dawley or Wistar rats. This form was variably expressed in livers of male Sprague Dawley and Wistar rats. Hepatic debrisoquine 4-hydroxylase activity was markedly reduced in female and male Dark Agouti rats as compared to Sprague Dawley or Wistar rats and correlated (r = 0.88; P < 0.001) with the hepatic CYP2D2 content. Recombinant CYP2D2 was 18-fold more active at catalysing the 4-hydroxylation of debrisoquine than CYP2D1. Furthermore, quinine markedly inhibited CYP2D2-mediated debrisoquine and metoprolol oxidation, while quinidine, its diastereoisomer, inhibited the reactions to a lesser extent. In conclusion, these results show that impaired debrisoquine 4-hydroxylase activity in the female Dark Agouti rat is due to low levels of CYP2D2.
Schulz-Utermoehl
T
T
Section on Clinical Pharmacology, Division of Medicine, Imperial College School of Medicine, London, UK.
Bennett
A J
AJ
Ellis
S W
SW
Tucker
G T
GT
Boobis
A R
AR
Edwards
R J
RJ
D000818:Animals; D000906:Antibodies / Q000276:immunology; D001189:Aryl Hydrocarbon Hydroxylases; D019389:Cytochrome P-450 CYP2D6 / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000276:immunology; D005260:Female; D008099:Liver / Q000201:enzymology; D008297:Male; D011110:Polymorphism, Genetic; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D013045:Species Specificity
D000906:Antibodies; D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; C119985:Cyp2d2 protein, rat; D019389:Cytochrome P-450 CYP2D6