Silva Josiane do Nascimento, Zampieri Thais T, Vieira Henrique R, Frazao Renata
Department of Anatomy, Instituto de Ciencias Biomedicas, Universidade de Sao Paulo.
Department of Anatomy, Instituto de Ciencias Biomedicas, Universidade de Sao Paulo;
J Vis Exp. 2023 Mar 17(193). doi: 10.3791/64989.
Kisspeptins are essential for the maturation of the hypothalamic-pituitary-gonadal (HPG) axis and fertility. Hypothalamic kisspeptin neurons located in the anteroventral periventricular nucleus and rostral periventricular nucleus, as well as the arcuate nucleus of the hypothalamus, project to gonadotrophin-releasing hormone (GnRH) neurons, among other cells. Previous studies have demonstrated that kisspeptin signaling occurs through the Kiss1 receptor (Kiss1r), ultimately exciting GnRH neuron activity. In humans and experimental animal models, kisspeptins are sufficient for inducing GnRH secretion and, consequently, luteinizing hormone (LH) and follicle stimulant hormone (FSH) release. Since kisspeptins play an essential role in reproductive functions, researchers are working to assess how the intrinsic activity of hypothalamic kisspeptin neurons contributes to reproduction-related actions and identify the primary neurotransmitters/neuromodulators capable of changing these properties. The whole-cell patch-clamp technique has become a valuable tool for investigating kisspeptin neuron activity in rodent cells. This experimental technique allows researchers to record and measure spontaneous excitatory and inhibitory ionic currents, resting membrane potential, action potential firing, and other electrophysiological properties of cell membranes. In the present study, crucial aspects of the whole-cell patch-clamp technique, known as electrophysiological measurements that define hypothalamic kisspeptin neurons, and a discussion of relevant issues about the technique, are reviewed.
kisspeptins对下丘脑 - 垂体 - 性腺（HPG）轴的成熟和生育能力至关重要。位于室周前腹侧核、室周吻侧核以及下丘脑弓状核的下丘脑kisspeptin神经元，除了投射到其他细胞外，还投射到促性腺激素释放激素（GnRH）神经元。先前的研究表明，kisspeptin信号通过Kiss1受体（Kiss1r）传递，最终激发GnRH神经元的活性。在人类和实验动物模型中，kisspeptins足以诱导GnRH分泌，从而导致黄体生成素（LH）和卵泡刺激素（FSH）释放。由于kisspeptins在生殖功能中起着至关重要的作用，研究人员正在努力评估下丘脑kisspeptin神经元的内在活性如何促进与生殖相关的行为，并确定能够改变这些特性的主要神经递质/神经调质。全细胞膜片钳技术已成为研究啮齿动物细胞中kisspeptin神经元活性的重要工具。这种实验技术使研究人员能够记录和测量自发的兴奋性和抑制性离子电流、静息膜电位、动作电位发放以及细胞膜的其他电生理特性。在本研究中，回顾了全细胞膜片钳技术的关键方面，即定义下丘脑kisspeptin神经元的电生理测量方法，并讨论了有关该技术的相关问题。
