Constantin Stephanie, Wray Susan
Cellular and Developmental Neurobiology Section, National Institute of Neurological Disorders and Stroke/National Institutes of Health, Bethesda, Maryland 20892-3703.
Endocrinology. 2016 Aug;157(8):3197-212. doi: 10.1210/en.2016-1064. Epub 2016 Jun 30.
GnRH neurons are regulated by hypothalamic kisspeptin neurons. Recently, galanin was identified in a subpopulation of kisspeptin neurons. Although the literature thoroughly describes kisspeptin activation of GnRH neurons, little is known about the effects of galanin on GnRH neurons. This study investigated whether galanin could alter kisspeptin signaling to GnRH neurons. GnRH cells maintained in explants, known to display spontaneous calcium oscillations, and a long-lasting calcium response to kisspeptin-10 (kp-10), were used. First, transcripts for galanin receptors (GalRs) were examined. Only GalR1 was found in GnRH neurons. A series of experiments was then performed to determine the action of galanin on kp-10 activated GnRH neurons. Applied after kp-10 activation, galanin 1-16 (Gal1-16) rapidly suppressed kp-10 activation. Applied with kp-10, Gal1-16 prevented kp-10 activation until its removal. To determine the mechanism by which galanin inhibited kp-10 activation of GnRH neurons, Gal1-16 and galanin were applied to spontaneously active GnRH neurons. Both inhibited GnRH neuronal activity, independent of GnRH neuronal inputs. This inhibition was mimicked by a GalR1 agonist but not by GalR2 or GalR2/3 agonists. Although Gal1-16 inhibition relied on Gi/o signaling, it was independent of cAMP levels but sensitive to blockers of G protein-coupled inwardly rectifying potassium channels. A newly developed bioassay for GnRH detection showed Gal1-16 decreased the kp-10-evoked GnRH secretion below detection threshold. Together, this study shows that galanin is a potent regulator of GnRH neurons, possibly acting as a physiological break to kisspeptin excitation.
促性腺激素释放激素(GnRH)神经元受下丘脑吻素神经元调控。最近,在一部分吻素神经元中发现了甘丙肽。尽管文献对吻素激活GnRH神经元已有详尽描述,但关于甘丙肽对GnRH神经元的作用却知之甚少。本研究调查了甘丙肽是否会改变吻素向GnRH神经元的信号传导。实验使用了维持在外植体中的GnRH细胞,已知这些细胞会显示出自发性钙振荡以及对吻素-10(kp-10)的持久钙反应。首先,检测了甘丙肽受体(GalRs)的转录本。在GnRH神经元中仅发现了GalR1。随后进行了一系列实验以确定甘丙肽对kp-10激活的GnRH神经元的作用。在kp-10激活后施加甘丙肽1-16(Gal1-16),可迅速抑制kp-10激活。与kp-10同时施加时,Gal1-16可阻止kp-10激活,直至其被去除。为了确定甘丙肽抑制GnRH神经元kp-10激活的机制,将Gal1-16和甘丙肽应用于自发活动的GnRH神经元。二者均抑制GnRH神经元活动,且与GnRH神经元输入无关。这种抑制作用可被GalR1激动剂模拟,但不能被GalR2或GalR2/3激动剂模拟。尽管Gal1-16的抑制作用依赖于Gi/o信号传导,但它与环磷酸腺苷(cAMP)水平无关,却对G蛋白偶联内向整流钾通道阻滞剂敏感。一种新开发的用于GnRH检测的生物测定法显示,Gal1-16可使kp-10诱发的GnRH分泌降至检测阈值以下。总之,本研究表明甘丙肽是GnRH神经元的有效调节剂,可能作为吻素兴奋的生理制动因素发挥作用。