Kisspeptin1
(product of the kiss1 gene) is the
key neuropeptide that gates puberty and maintains fertility by regulating the gonadotropin-releasing
hormone (GnRH) neuronal system in mammals. Inactivating mutations in kiss1 and
the kisspeptin receptor (GPR54/KiSS1R) are associated with pubertal failure and
infertility. Kiss2, a paralogous gene
for kiss1, has been recently
identified in several vertebrates including zebrafish. Using our transgenic
zebrafish model system in which the GnRH3 promoter drives expression of emerald
green fluorescent protein, we investigated the effects of kisspeptins on
development of the GnRH neuronal system during embryogenesis and on electrical
activity during adulthood. Quantitative PCR showed detectable levels of kiss1 and kiss2 mRNA by 1 day post fertilization, increasing throughout
embryonic and larval development. Early treatment with kisspeptin1 or
kisspeptin2 showed that both kisspeptins stimulated proliferation of trigeminal
GnRH3 neurons located in the peripheral nervous system. However, only kisspeptin1,
but not kisspeptin2, stimulated proliferation of the terminal nerve and
hypothalamic populations of GnRH3 neurons in the central nervous system.
Immunohistochemical analysis of synaptic vesicle protein 2 suggested that kisspeptin1,
but not kisspeptin2, increased synaptic contacts along the terminal nerve-GnRH3
neuronal processes during embryogenesis. In intact brain of adult zebrafish,
whole-cell patch clamp recordings of GnRH3 neurons from the preoptic area and
hypothalamus revealed opposite effects of kisspeptin1 and kisspeptin2 on
spontaneous action potential firing frequency and membrane potential. Kisspeptin1
increased spike frequency and depolarized membrane potential, whereas
kisspeptin2 suppressed spike frequency and hyperpolarized membrane potential.
We conclude that in zebrafish, kisspeptin1 is the primary stimulator of GnRH3
neuronal development in the embryo and an activator of stimulating
hypophysiotropic neuron activities in the adult, while kisspeptin2 plays an additional
role in stimulating embryonic development of the trigeminal neuronal
population, but is an RFamide inhibitor of electrical activity of
hypophysiotropic GnRH3 neurons in the adult.