Neurosteroid synthesis by cytochrome P450-containing systems localized in the rat brain hippocampal neurons: N-methyl-D-aspartate and calcium-dependent synthesis

Tetsuya Kimoto, Tomokazu Tsurugizawa, Yoichiro Ohta, Jun'ya Makino, Hiroomi Tamura, Yasushi Hojo, Norio Takata, Suguru Kawato

Research output: Contribution to journalArticle

199 Citations (Scopus)

Abstract

Neurosteroidogenesis has not been well elucidated due to the very low level of steroidogenic proteins in the brain. Here we report the first demonstration of the neuronal localization of neurosteroidogenic systems as well as the regulation of neurosteroidogenic activity in the adult rat hippocampus. Significant localization of cytochrome P450scc was observed in pyramidal neurons and granule neurons by means of immunohistochemical staining of slices. We also observed the colocalization, in hippocampal neurons, of P450scc with redox partners, hydroxysteroid sulfotransferase and steroidogenic acute regulatory protein. The distributions of astroglial cells and oligodendroglial cells showed very different patterns from that of the P450scc-containing cells. The expression of P450scc, redox partners, the sulfotransferase, and steroidogenic acute regulatory protein was also confirmed by Western blot analysis. The process of active neurosteroidogenesis was stimulated by exposing neurons to N-methyl-D-aspartate. Upon stimulation with N-methyl-D-aspartate, Ca2+ influx through the N-methyl-D-aspartate subtype of glutamate receptors occurred, and significant net production of pregnenolone and pregnenolone sulfate was observed in the hippocampus. This neurosteroid production was considerably suppressed by the addition of antagonists of N-methyl-D-aspartate receptors, by Ca2+ depletion, or by the addition of an inhibitor of P450scc. Upon stimulation with N-methyl-D-aspartate, the processing of full-length steroidogenic acute regulatory protein (37-kDa) to the truncated 30-kDa steroidogenic acute regulatory protein was observed. Taken together, these observations imply that hippocampal neurons synthesize neurosteroids. This synthesis may be stimulated and regulated by glutamate-mediated synaptic communication.

Original languageEnglish
Pages (from-to)3578-3589
Number of pages12
JournalEndocrinology
Volume142
Issue number8
DOIs
Publication statusPublished - 2001

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N-Methylaspartate
Cytochrome P-450 Enzyme System
Neurotransmitter Agents
Calcium
Neurons
Brain
Oxidation-Reduction
Hippocampus
Cholesterol Side-Chain Cleavage Enzyme
Sulfotransferases
Pregnenolone
Pyramidal Cells
Glutamate Receptors
N-Methyl-D-Aspartate Receptors
Glutamic Acid
Western Blotting
Communication
Staining and Labeling
steroidogenic acute regulatory protein
Proteins

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Neurosteroid synthesis by cytochrome P450-containing systems localized in the rat brain hippocampal neurons : N-methyl-D-aspartate and calcium-dependent synthesis. / Kimoto, Tetsuya; Tsurugizawa, Tomokazu; Ohta, Yoichiro; Makino, Jun'ya; Tamura, Hiroomi; Hojo, Yasushi; Takata, Norio; Kawato, Suguru.

In: Endocrinology, Vol. 142, No. 8, 2001, p. 3578-3589.

Research output: Contribution to journalArticle

Kimoto, Tetsuya ; Tsurugizawa, Tomokazu ; Ohta, Yoichiro ; Makino, Jun'ya ; Tamura, Hiroomi ; Hojo, Yasushi ; Takata, Norio ; Kawato, Suguru. / Neurosteroid synthesis by cytochrome P450-containing systems localized in the rat brain hippocampal neurons : N-methyl-D-aspartate and calcium-dependent synthesis. In: Endocrinology. 2001 ; Vol. 142, No. 8. pp. 3578-3589.
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