Expression of cytochrome P450 side-chain cleavage enzyme mRNA and production of pregnenolone in the brain of the red-bellied newt Cynops pyrrhogaster

Minoru Takase, Shogo Haraguchi, Itaru Hasunuma, Sakae Kikuyama, Kazuyoshi Tsutsui

    Research output: Contribution to journalArticle

    17 Citations (Scopus)

    Abstract

    It is becoming clear that the vertebrate brain has the capability of forming steroids de novo, the so-called " neurosteroids" To understand neurosteroidogenesis in the brain, it is essential to demonstrate the formation of pregnenolone, a main precursor of neurosteroids. In amphibians, the pregnenolone formation from cholesterol is still unclear, although the brain accumulates pregnenolone, pregnenolone sulfate and 7α-hydroxypregnenolone. This study was addressed to obtain basic information about pregnenolone formation in the newt brain. Firstly, we demonstrated that the newt brain produces pregnenolone from cholesterol. Subsequently, cDNA encoding cytochrome P450 side-chain cleavage enzyme (P450scc), a key steroidogenic enzyme catalyzing pregnenolone formation, was isolated from the newt. The sequence analysis showed that the isolated P450scc cDNA contained a putative coding region consisting of 1569. bp, which encoded 523 amino acids. The steroid- and heme-binding domains of P450scc were highly shared in amino acids among vertebrates. RT-PCR analysis amplified the authentic fragment corresponding to newt P450scc showed its transcription in the brain. However, the transcription level in the brain was lower than those of the gonad and the kidney including adrenals. The restricted cells in the four major regions of the newt brain, such as the telencephalon, diencephalon, mesencephalon, and rhombencephalon, were demonstrated to express P450scc transcripts by RT-PCR and in situ hybridization. Taken together, these results indicate that the newt brain expresses P450scc mRNA and produces pregnenolone from cholesterol.

    Original languageEnglish
    Pages (from-to)468-474
    Number of pages7
    JournalGeneral and Comparative Endocrinology
    Volume170
    Issue number3
    DOIs
    Publication statusPublished - 2011 Feb 1

    Fingerprint

    pregnenolone
    Salamandridae
    Pregnenolone
    salamanders and newts
    cytochrome P-450
    Cytochrome P-450 Enzyme System
    brain
    Messenger RNA
    Brain
    Enzymes
    enzymes
    Cholesterol
    cholesterol
    steroids
    Neurotransmitter Agents
    Vertebrates
    Complementary DNA
    transcription (genetics)
    reverse transcriptase polymerase chain reaction
    Steroids

    Keywords

    • Brain
    • Cytochrome P450scc
    • Neurosteroids
    • Newt
    • Pregnenolone

    ASJC Scopus subject areas

    • Endocrinology

    Cite this

    Expression of cytochrome P450 side-chain cleavage enzyme mRNA and production of pregnenolone in the brain of the red-bellied newt Cynops pyrrhogaster. / Takase, Minoru; Haraguchi, Shogo; Hasunuma, Itaru; Kikuyama, Sakae; Tsutsui, Kazuyoshi.

    In: General and Comparative Endocrinology, Vol. 170, No. 3, 01.02.2011, p. 468-474.

    Research output: Contribution to journalArticle

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