Zhang Chunguang, Bosch Martha A, Qiu Jian, Rønnekleiv Oline K, Kelly Martin J
Department of Physiology and Pharmacology (C.Z., M.A.B., J.Q., O.K.R., M.J.K.), Oregon Health and Science University, Portland, Oregon 97239; and Division of Neuroscience (O.K.R., M.J.K.), Oregon National Primate Research Center, Oregon Health and Science University, Beaverton, Oregon 97006.
Mol Endocrinol. 2015 Apr;29(4):518-27. doi: 10.1210/me.2014-1392. Epub 2015 Mar 3.
In vitro slice studies have revealed that there are significant differences in the spontaneous firing activity between anteroventral periventricular/periventricular preoptic nucleus (AVPV/PeN) and arcuate nucleus (ARC) kisspeptin (Kiss1) neurons in females. Although both populations express similar endogenous conductances, we have discovered that AVPV/PeN Kiss1 neurons express a subthreshold, persistent sodium current (INaP) that dramatically alters their firing activity. Based on whole-cell recording of Kiss1-Cre-green fluorescent protein (GFP) neurons, INaP was 4-fold greater in AVPV/PeN vs ARC Kiss1 neurons. An LH surge-producing dose of 17β-estradiol (E2) that increased Kiss1 mRNA expression in the AVPV/PeN, also augmented INaP in AVPV/PeN neurons by 2-fold. Because the activation threshold for INaP was close to the resting membrane potential (RMP) of AVPV/PeN Kiss1 neurons (-54 mV), it rendered them much more excitable and spontaneously active vs ARC Kiss1 neurons (RMP = -66 mV). Single-cell RT-PCR revealed that AVPV/PeN Kiss1 neurons expressed the requisite sodium channel α-subunit transcripts, NaV1.1, NaV1.2, and NaV1.6 and β subunits, β2 and β4. Importantly, NaV1.1α and -β2 transcripts in AVPV/PeN, but not ARC, were up-regulated 2- to 3-fold by a surge-producing dose of E2, similar to the transient calcium current channel subunit Cav3.1. The transient calcium current collaborates with INaP to generate burst firing, and selective blockade of INaP by riluzole significantly attenuated rebound burst firing and spontaneous activity. Therefore, INaP appears to play a prominent role in AVPV/PeN Kiss1 neurons to generate spontaneous, repetitive burst firing, which is required for the high-frequency-stimulated release of kisspeptin for exciting GnRH neurons and potentially generating the GnRH surge.
体外脑片研究表明,雌性动物前腹侧室周/室周视前核(AVPV/PeN)和弓状核(ARC)的促性腺激素释放激素神经元(Kiss1)在自发放电活动上存在显著差异。尽管这两类神经元群体都表达相似的内源性电导,但我们发现,AVPV/PeN Kiss1神经元表达一种阈下持续性钠电流(INaP),这极大地改变了它们的放电活动。基于对Kiss1-Cre绿色荧光蛋白(GFP)神经元的全细胞记录,AVPV/PeN Kiss1神经元中的INaP比ARC Kiss1神经元中的高4倍。能引起促黄体生成素激增的17β-雌二醇(E2)剂量,可增加AVPV/PeN中Kiss1 mRNA的表达,同时也使AVPV/PeN神经元中的INaP增加了2倍。由于INaP的激活阈值接近AVPV/PeN Kiss1神经元的静息膜电位(RMP)(-54 mV),与ARC Kiss1神经元(RMP = -66 mV)相比,这使得它们更易兴奋且自发放电。单细胞逆转录PCR显示,AVPV/PeN Kiss1神经元表达必需的钠通道α亚基转录本NaV1.1、NaV1.2和NaV1.6以及β亚基β2和β4。重要的是,能引起激增的E2剂量使AVPV/PeN而非ARC中的NaV1.1α和-β2转录本上调了2至3倍,类似于瞬时钙电流通道亚基Cav3.1。瞬时钙电流与INaP协同产生爆发式放电,利鲁唑对INaP的选择性阻断显著减弱了反弹爆发式放电和自发放电。因此,INaP似乎在AVPV/PeN Kiss1神经元产生自发、重复性爆发式放电中起重要作用,这是促性腺激素释放激素高频刺激释放以兴奋促性腺激素释放激素神经元并可能产生促性腺激素释放激素激增所必需的。