Cholesterol metabolism

the main pathway acting downstream of cytochrome P450 oxidoreductase in skeletal development of the limb

Katy Schmidt, Catherine Hughes, J A Chudek, Simon Robert Goodyear, Richard Malcolm Aspden, Richard Talbot, Thomas E Gundersen, Rune Blomhoff, Colin Henderson, C Roland Wolf, Cheryll Tickle

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Cytochrome P450 oxidoreductase (POR) is the obligate electron donor for all microsomal cytochrome P450 enzymes, which catalyze the metabolism of a wide spectrum of xenobiotic and endobiotic compounds. Point mutations in POR have been found recently in patients with Antley-Bixler-like syndrome, which includes limb skeletal defects. In order to study P450 function during limb and skeletal development, we deleted POR specifically in mouse limb bud mesenchyme. Forelimbs and hind limbs in conditional knockout (CKO) mice were short with thin skeletal elements and fused joints. POR deletion occurred earlier in forelimbs than in hind limbs, leading additionally to soft tissue syndactyly and loss of wrist elements and phalanges due to changes in growth, cell death, and skeletal segmentation. Transcriptional analysis of E12.5 mouse forelimb buds demonstrated the expression of P450s involved in retinoic acid, cholesterol, and arachidonic acid metabolism. Biochemical analysis of CKO limbs confirmed retinoic acid excess. In CKO limbs, expression of genes throughout the whole cholesterol biosynthetic pathway was upregulated, and cholesterol deficiency can explain most aspects of the phenotype. Thus, cellular POR-dependent cholesterol synthesis is essential during limb and skeletal development. Modulation of P450 activity could contribute to susceptibility of the embryo and developing organs to teratogenesis.
Original languageEnglish
Pages (from-to)2716-2729
Number of pages14
JournalMolecular and Cellular Biology
Volume29
Issue number10
DOIs
Publication statusPublished - May 2009

Fingerprint

Cytochrome P-450 Enzyme System
Oxidoreductases
Extremities
Cholesterol
Forelimb
Tretinoin
Antley-Bixler Syndrome Phenotype
Teratogenesis
Syndactyly
Limb Buds
Biosynthetic Pathways
Mesoderm
Xenobiotics
Wrist
Point Mutation
Arachidonic Acid
Knockout Mice
Cell Death
Embryonic Structures
Joints

Keywords

  • Animals
  • Antley-Bixler Syndrome Phenotype
  • Bone and Bones
  • Cholesterol
  • Chondrogenesis
  • Embryo, Mammalian
  • Extremities
  • Humans
  • Limb Deformities, Congenital
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • NADPH-Ferrihemoprotein Reductase
  • Oligonucleotide Array Sequence Analysis
  • Phenotype
  • Signal Transduction

Cite this

Cholesterol metabolism : the main pathway acting downstream of cytochrome P450 oxidoreductase in skeletal development of the limb. / Schmidt, Katy; Hughes, Catherine; Chudek, J A; Goodyear, Simon Robert; Aspden, Richard Malcolm; Talbot, Richard; Gundersen, Thomas E; Blomhoff, Rune; Henderson, Colin; Wolf, C Roland; Tickle, Cheryll.

In: Molecular and Cellular Biology, Vol. 29, No. 10, 05.2009, p. 2716-2729.

Research output: Contribution to journalArticle

Schmidt, K, Hughes, C, Chudek, JA, Goodyear, SR, Aspden, RM, Talbot, R, Gundersen, TE, Blomhoff, R, Henderson, C, Wolf, CR & Tickle, C 2009, 'Cholesterol metabolism: the main pathway acting downstream of cytochrome P450 oxidoreductase in skeletal development of the limb', Molecular and Cellular Biology, vol. 29, no. 10, pp. 2716-2729. https://doi.org/10.1128/MCB.01638-08
Schmidt, Katy ; Hughes, Catherine ; Chudek, J A ; Goodyear, Simon Robert ; Aspden, Richard Malcolm ; Talbot, Richard ; Gundersen, Thomas E ; Blomhoff, Rune ; Henderson, Colin ; Wolf, C Roland ; Tickle, Cheryll. / Cholesterol metabolism : the main pathway acting downstream of cytochrome P450 oxidoreductase in skeletal development of the limb. In: Molecular and Cellular Biology. 2009 ; Vol. 29, No. 10. pp. 2716-2729.
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AU - Chudek, J A

AU - Goodyear, Simon Robert

AU - Aspden, Richard Malcolm

AU - Talbot, Richard

AU - Gundersen, Thomas E

AU - Blomhoff, Rune

AU - Henderson, Colin

AU - Wolf, C Roland

AU - Tickle, Cheryll

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N2 - Cytochrome P450 oxidoreductase (POR) is the obligate electron donor for all microsomal cytochrome P450 enzymes, which catalyze the metabolism of a wide spectrum of xenobiotic and endobiotic compounds. Point mutations in POR have been found recently in patients with Antley-Bixler-like syndrome, which includes limb skeletal defects. In order to study P450 function during limb and skeletal development, we deleted POR specifically in mouse limb bud mesenchyme. Forelimbs and hind limbs in conditional knockout (CKO) mice were short with thin skeletal elements and fused joints. POR deletion occurred earlier in forelimbs than in hind limbs, leading additionally to soft tissue syndactyly and loss of wrist elements and phalanges due to changes in growth, cell death, and skeletal segmentation. Transcriptional analysis of E12.5 mouse forelimb buds demonstrated the expression of P450s involved in retinoic acid, cholesterol, and arachidonic acid metabolism. Biochemical analysis of CKO limbs confirmed retinoic acid excess. In CKO limbs, expression of genes throughout the whole cholesterol biosynthetic pathway was upregulated, and cholesterol deficiency can explain most aspects of the phenotype. Thus, cellular POR-dependent cholesterol synthesis is essential during limb and skeletal development. Modulation of P450 activity could contribute to susceptibility of the embryo and developing organs to teratogenesis.

AB - Cytochrome P450 oxidoreductase (POR) is the obligate electron donor for all microsomal cytochrome P450 enzymes, which catalyze the metabolism of a wide spectrum of xenobiotic and endobiotic compounds. Point mutations in POR have been found recently in patients with Antley-Bixler-like syndrome, which includes limb skeletal defects. In order to study P450 function during limb and skeletal development, we deleted POR specifically in mouse limb bud mesenchyme. Forelimbs and hind limbs in conditional knockout (CKO) mice were short with thin skeletal elements and fused joints. POR deletion occurred earlier in forelimbs than in hind limbs, leading additionally to soft tissue syndactyly and loss of wrist elements and phalanges due to changes in growth, cell death, and skeletal segmentation. Transcriptional analysis of E12.5 mouse forelimb buds demonstrated the expression of P450s involved in retinoic acid, cholesterol, and arachidonic acid metabolism. Biochemical analysis of CKO limbs confirmed retinoic acid excess. In CKO limbs, expression of genes throughout the whole cholesterol biosynthetic pathway was upregulated, and cholesterol deficiency can explain most aspects of the phenotype. Thus, cellular POR-dependent cholesterol synthesis is essential during limb and skeletal development. Modulation of P450 activity could contribute to susceptibility of the embryo and developing organs to teratogenesis.

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KW - Extremities

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KW - Limb Deformities, Congenital

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KW - Mice, Inbred C57BL

KW - Mice, Knockout

KW - NADPH-Ferrihemoprotein Reductase

KW - Oligonucleotide Array Sequence Analysis

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KW - Signal Transduction

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