MOLECULAR DYNAMICS APPROACH TO GNRH PULSATILITY

描述（改编自摘要）： 下丘脑一小部分神经元GnRH的脉冲性释放 是哺乳动物生殖功能的驱动力。这种分泌 似乎依赖于细胞内游离钙的振荡 是由间歇性的钙离子通过电压流入引起的 操纵膜钙通道。最近的证据也表明 GnRH表达的变化以偶发方式发生。为了获得 对这些动态波动的机械理解，新的细胞 和更接近于“实时”的分子策略 需要蜂窝事件。私家侦探和他的同事们 策略，使多个测量GnRH基因表达（通过 荧光素酶报告基因活性成像）、GnRH释放（通过 膜染料摄取和再循环），钙振荡（Fura-2 成像），以及这些在活GT-1细胞中的组合。使用这些 工具，具体目标是：1）阐明分子动力学 在活的GT-1细胞中的GnRH基因表达是 永生化GnRH神经元; 2）探索分泌的细胞基础 单个GnRH神经元的脉动性;和3）建立可能的 GnRH分泌、基因表达和钙之间的关系 在同一个活细胞中的动力学

DESCRIPTION (Adapted from abstract): The pulsatile release of GnRH in a small subset of hypothalamic neurons is a driving force for reproductive function in mammals. This secretion appears to be dependent on oscillations in intracellular free calcium that are derived from intermittent calcium influx through voltage- operated membrane calcium channels. Recent evidence also suggests changes in GnRH expression occur in an episodic manner. In order to gain a mechanistic understanding of these dynamic fluctuations, new cellular and molecular strategies corresponding more closely to "real-time" cellular events are required. The PI and his colleagues have developed strategies to make multiple measurements of GnRH gene expression (via luciferase reporter gene activity imaging), GnRH release (by changes in membrane dye uptake and recycling), calcium oscillations (Fura-2 imaging), and combinations of these in living GT-1 cells. Using these tools, the specific aims are to: 1) elucidate the molecular dynamics of GnRH gene expression in living GT-1 cells are representatives of immortalized GnRH neurons; 2) explore the cellular basis for secretory pulsatility in individual GnRH neurons; and 3) establish possible relationships among GnRH secretion, gene expression, and calcium dynamics in the same, living cells.