Cytochrome p450 side-chain cleavage enzyme in the cerebellar purkinje neuron and its neonatal change in rats

Kazuyoshi Ukena, Mariko Usui, Chie Kohchi, Kazuyoshi Tsutsui

Research output: Contribution to journalArticle

150 Citations (Scopus)

Abstract

Neurosteroids are de novo synthesized in the nervous system through mechanisms at least partly independent of peripheral steroidogenic glands. However, the concept of neurosteroidogenesis in neurons is not clear in mammalian brains. The present study identified the presence of cytochrome P450scc in the rat Purkinje cell, a typical cerebellar neuron. Immunohistochemical analysis with the antibody against the purified bovine adrenal P450scc showed an immunoreaction restricted to somata and dendrites of the Purkinje cells in adult cerebella. Preadsorbing the antibody with P450scc resulted in a complete absence of the immunoreaction. The antibody against inositol triphosphate receptor, a marker of the Purkinje cell, recognized P450scc-immunoreactive cerebellar cells that showed no immunoreaction with glial fibrillary acidic protein, a specific marker of glial cells. Expression of the P450scc-like protein in the cerebellum was verified by Western blot analysis, and cerebellar P450scc messenger RNA, by RT-PCR analysis in adulthood. On the other hand, P450scc-immunoreactive cells were found to scatter throughout the cerebellum at 0 day of age, before the differentiation of the first Purkinje cells, while the site of expression of this protein was localized only in somata of Purkinje cells at 3 days of age. Immunoreactive dendrites of the Purkinje cell spread into the molecular layer during neonatal development concurrently with its maturation. The intensity of the immunoreaction did not change during neonatal life. Expression of the cerebellar P450scc messenger RNA was also detected after birth, and the level was almost constant during neonatal life. A specific RIA indicated that the pregnenolone concentration was unexpectedly high at 0 day and decreased until 7 days. The total amount of pregnenolone in the cerebellum was almost constant from 0-7 days and increased during 7-21 days concurrently with the cerebellar development. In contrast, the pregnenolone sulfate ester level was low and did not significantly change among the developmental stages. These results suggest that steroidogenic enzyme P450scc appears in the rat Purkinje cell immediately after its differentiation. The expression of this enzyme may remain during neonatal development and in adulthood.

Original languageEnglish
Pages (from-to)137-147
Number of pages11
JournalEndocrinology
Volume139
Issue number1
DOIs
Publication statusPublished - 1998
Externally publishedYes

Fingerprint

Purkinje Cells
Cytochrome P-450 Enzyme System
Cerebellum
Enzymes
Pregnenolone
Carisoprodol
Dendrites
Antibodies
Cholesterol Side-Chain Cleavage Enzyme
Inositol 1,4,5-Trisphosphate Receptors
Neurons
Messenger RNA
Glial Fibrillary Acidic Protein
Neuroglia
Nervous System
Neurotransmitter Agents
Esters
Proteins
Western Blotting
Parturition

ASJC Scopus subject areas

  • Endocrinology

Cite this

Cytochrome p450 side-chain cleavage enzyme in the cerebellar purkinje neuron and its neonatal change in rats. / Ukena, Kazuyoshi; Usui, Mariko; Kohchi, Chie; Tsutsui, Kazuyoshi.

In: Endocrinology, Vol. 139, No. 1, 1998, p. 137-147.

Research output: Contribution to journalArticle

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