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Crosstalk between kisspeptin and gonadotropin-inhibitory hormone in the silence of puberty: preclinical evidence from a calcium signaling study

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TAYLOR & FRANCIS LTD
DOI: 10.1080/10799893.2022.2125014

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Kisspeptin; gonadotropin-inhibitory hormone; calcium signaling; puberty

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This study investigated the effects of kisspeptin and GnIH on calcium signaling using rHypoE-8 cells as a model. The results showed that kisspeptin significantly increased intracellular calcium levels in rHypoE-8 cells, while GnIH did not cause significant changes in basal calcium levels but attenuated kisspeptin-induced calcium transients. These findings suggest that kisspeptin and GnIH may modulate the function of the HPG axis by regulating kisspeptin neurons.
Kisspeptin and gonadotropin-inhibitory hormone (GnIH) are among suggested neuroendocrine modulators of reproductive function. Intracellular calcium signaling is a critical component in the regulation of a variety of physiological and pathological processes including neurotransmitter release, and, therefore, can be used as signaling indicator for investigating the involvement of kisspeptin, GnIH, and gonadotropin-releasing hormone (GnRH) release. Hence, this study investigated the effects of kisspeptin and GnIH on calcium signaling using immortalized hypothalamic cells (rHypoE-8) as a model. Kisspeptin neurons were loaded with the ratiometric calcium dye (Fura-2 AM, 1 mu mol) and intracellular free calcium ([Ca2+](i)) responses were quantified using digital fluorescence imaging system. Kisspeptin-10 (100, 300, and 1000 nM) caused a significant increase in [Ca2+](i) in rHypoE-8 cells (n = 58, n = 64, and n = 49, respectively, p < 0.001). The kisspeptin receptor antagonist, P234, inhibited the calcium responses to kisspeptin (p < 0.001, n = 32). GnIH (100 and 1000 nM), alone, did not cause any significant change in the mean basal [Ca2+](i) levels in kisspeptin cells, but GnIH attenuated the kisspeptin-evoked [Ca2+](i) transients (n = 47, p < 0.001). This novel findings of [Ca2+](i) signaling in in vitro setting implicate that kisspeptin and GnIH may exert their effects on hypothalamus-pituitary-gonadal (HPG) axis by modulating kisspeptin neurons. These results also implicate that kisspeptin neurons may have an autocrine regulation.

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