Expression patterns of mouse and human CYP orthologs (families 1–4) during development and in different adult tissues

doi:10.1016/j.abb.2005.02.001

Archives of Biochemistry and Biophysics

Volume 436, Issue 1, 1 April 2005, Pages 50-61

https://doi.org/10.1016/j.abb.2005.02.001 Get rights and content

Abstract

The present study compared the relative expression pattern of 10 orthologous CYP forms from families 1–4 in cDNA panels of human and mouse fetal and adult tissues. Except for CYP1A2, all of these CYPs exhibited specific patterns of expression during mouse ontogeny, suggesting possible involvement in development. Cyp1a1 and Cyp2r1 were the only two of the orthologs to be expressed only in the E7 mouse; Cyp2s1 was expressed in all stages, including E7, while Cyp2e1 appeared only at E17. Highest expression of the individual CYPs in the different late term human fetal tissues was: thymus, CYP1B1 and CYP2U1; liver, CYP2E1; brain, CYP2R1, CYP1A1 and CYP4X1; and lung, CYP4B1 and CYP2W1. In general, the level of individual human CYP transcripts was lower in the fetal than the corresponding adult tissues. The pattern of expression in adult mouse and human tissues was fairly similar for CYP1A1, CYP1A2, CYP1B1, CYP2S1, and CYP2U1 orthologs.

Section snippets

Materials and methods

The PCR reagents (dNTPs, Platinum Taq DNA polymerase, and MgCl₂) were obtained from Invitrogen, Carlsbad, CA. Normalized MTC panels were purchased from Clontech, Palo Alto, CA. All other chemicals used in the study were of analytical grade.

Results

A distinctive constitutive expression pattern of 10 orthologous CYP genes in human and mouse was observed using MTC panels, normalized against four different housekeeping genes, G3PDH, phospholipase A2, ribosomal protein S29, and β-actin (Clontech). The mouse panel contained cDNA from four different developmental stages and eight adult tissues of mouse. Human samples consisted of a panel of eight human fetal tissues and two adult human panels, each containing cDNA of eight tissues. The PCR were

Discussion

A wealth of information has been obtained on newly identified genes due to the completion of genome sequencing projects of multiple species. Many of the genes being revealed are related and similar in function. Hence, the delineation of such orthologous genes is significant in annotating functional and evolutionary aspects of comparative genomics [17]. The spatial and temporal expression patterns of these genes are determined by the response elements present in promoter sequences. A recent

Acknowledgments

This work was supported in part by NIH Grants 1R01 EY11095 and 2R01 ES03154, by the American Assistance Foundation National Glaucoma Research Grant G2000-021, and by an award from the Sandler Program for Asthma Research.

References (46)

D.R. Nelson
Arch. Biochem. Biophys.
(2003)
J.B. Cheng et al.
J. Biol. Chem.
(2003)
S.S. Chuang et al.
J. Biol. Chem.
(2004)
G. Smith et al.
Lancet
(2003)
M.S. Miller et al.
Fund. App. Toxicol.
(1996)
D. Choudhary et al.
Arch. Biochem. Biophys.
(2003)
T.A. Tatusova et al.
FEMS Microbiol. Lett.
(1999)
S.F. Altschul et al.
J. Mol. Biol.
(1990)
S.J. Yaffe et al.
Life Sci.
(1970)
J.V. Chithalen et al.
J. Lipid Res.
(2002)

D.F.V. Lewis et al.

Chem. Biol. Interact.

(2003)

J.B. Schenkman et al.

Proc. Indian natn. Sci. Acad. B

(2003)

D. Choudhary et al.

Drug Metab. Rev.

(2004)

D.R. Nelson et al.

Pharmacogenetics

(2004)

I. Stoilov et al.

Hum. Mol. Genet.

(1997)

R.T. Libby et al.

Science

(2003)

S. Rozen et al.

P.D. Siebert et al.

Biotechniques

(1993)

F. Sanger et al.

Proc. Natl. Acad. Sci. USA

(1977)

R.L. Tatusov et al.

BMC Bioinform.

(2003)

Y.V. Sun et al.

Nucleic Acids Res.

(2004)

H. Maraguchi et al.

Fungal Genet. Biol.

(2000)

G.-T. Kim et al.

Genes Dev.

(1998)

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Expression patterns of mouse and human CYP orthologs (families 1–4) during development and in different adult tissues

Abstract

Section snippets

Materials and methods

Results

Discussion

Acknowledgments

Arch. Biochem. Biophys.

J. Biol. Chem.

J. Biol. Chem.

Lancet

Fund. App. Toxicol.

Arch. Biochem. Biophys.

FEMS Microbiol. Lett.

J. Mol. Biol.

Life Sci.

J. Lipid Res.

Chem. Biol. Interact.

Proc. Indian natn. Sci. Acad. B

Drug Metab. Rev.

Pharmacogenetics

Hum. Mol. Genet.

Science

Biotechniques

Proc. Natl. Acad. Sci. USA

BMC Bioinform.

Nucleic Acids Res.

Fungal Genet. Biol.

Genes Dev.