Characterization and transcriptional profiles of Engraulis encrasicolus’ GnRH forms

The European anchovy Engraulis encrasicolus, a member of the Clupeiformes order, holds a great biological and economical importance. In the past, this species was mostly investigated with the aim of assessing its reproductive biology, trophic ecology, population dynamics and the relations existing with the physical environment. At present days, though, an almost complete lack of information afflicts its neuroendocrinology and reproductive physiology. The hypothalamic–pituitary–gonadal (HPG) axis at its highest levels was herein investigated. In this study, the gonadotropin-releasing hormone (GnRH), a neuropeptide underlying many reproduction-related processes, the most critical of which is the stimulation of gonadotropin synthesis and secretion from the pituitary gland, was cloned. Three forms (salmon GnRH, chicken-II GnRH and the species-specific type) were characterized in their full-length open-reading frames and, in accordance with other Clupeiformes species, the distinctive one was found to be the herring-type GnRH. We qualitatively and semiquantitatively evaluated the localizations of expressions and the temporal transcription patterns of the three GnRH forms in male and female specimens throughout their reproductive cycle as well as described their phylogeny with regard to teleost GnRH lineages, and, specifically, to other Clupeiformes species.

Selective Modulation of Voltage-Gated Calcium Channels in the Terminal Nerve Gonadotropin-Releasing Hormone Neurons of a Teleost, the Dwarf Gourami (Colisa lalia)

GnRH neurons in the terminal nerve (TN) have been suggested to function as a neuromodulatory system that regulates long-lasting changes in the animal behavior. Here we examined electrophysiological properties of TN-GnRH neurons in a teleost (dwarf gourami, Colisa lalia), focusing on the voltage-gated Ca2+ channels, which are thought to be coupled to several cellular events such as GnRH release. TN-GnRH neurons showed low-voltage activated (LVA) currents and three types of pharmacologically distinct high-voltage activated (HVA) currents. The L- and N-type currents constituted 30.7 ± 3.1 and 41.0 ± 3.9%, respectively, of HVA currents, which was recorded at the holding potential of −60 mV to inactivate the LVA currents. Although P/Q-type current was small and negligible, R-type current accounted for the remaining 23.6 ± 1.6% of HVA currents. Next we examined the possibility of Ca2+ channel modulation induced by GnRH released in a paracrine/autocrine manner. HVA currents of up to 40% was inhibited by the application of salmon GnRH, which is the same molecular species of GnRH as is synthesized by TN-GnRH neurons themselves. However, salmon GnRH had no measurable effects on LVA currents. The inhibition of HVA currents had a dose dependence (EC50 was 11.5 nm) and type specificity among different HVA currents; N- and R-type currents were preferentially inhibited, but L-type currents had by far lower sensitivity. The physiological significance of different Ca2+ influx pathways, and their paracrine/autocrine regulation mechanisms in TN-GnRH neurons are discussed.