Prolactin Increases the Synthesis of 7α-Hydroxypregnenolone, a Key Factor for Induction of Locomotor Activity, in Breeding Male Newts

Abstract

We recently found that the Japanese red-bellied newt, Cynops pyrrhogaster, actively produces 7α-hydroxypregnenolone, a previously undescribed amphibian neurosteroid. 7α-Hydroxypregnenolone stimulates locomotor activity of male newts. Locomotor activity of male newts increases during the breeding period as in other wild animals, but the molecular mechanism for such a change in locomotor activity is poorly understood. Here we show that the adenohypophyseal hormone prolactin (PRL) stimulates 7α-hydroxypregnenolone synthesis in the brain, thus increasing locomotor activity of breeding male newts. In this study, cytochrome P4507α (CYP7B), a steroidogenic enzyme catalyzing the formation of 7α-hydroxypregnenolone, was first identified to analyze seasonal changes in 7α-hydroxypregnenolone synthesis. Only males exhibited marked seasonal changes in 7α-hydroxypregnenolone synthesis and CYP7B expression in the brain, with a maximum level in the spring breeding period when locomotor activity of males increases. Subsequently we identified PRL as a key component of the mechanism regulating 7α-hydroxypregnenolone synthesis. Hypophysectomy decreased 7α-hydroxypregnenolone synthesis in the male brain, whereas administration of PRL but not gonadotropins to hypophysectomized males caused a dose-dependent increase in 7α-hydroxypregnenolone synthesis. To analyze the mode of PRL action, CYP7B and the receptor for PRL were localized in the male brain. PRL receptor was expressed in the neurons expressing CYP7B in the magnocellular preoptic nucleus. Thus, PRL appears to act directly on neurosteroidogenic magnocellular preoptic nucleus neurons to regulate 7α-hydroxypregnenolone synthesis, thus inducing seasonal locomotor changes in male newts. This is the first report describing the regulation of neurosteroidogenesis in the brain by an adenohypophyseal hormone in any vertebrate.