CARDIAC GLYCOSIDES FROM THE ADRENAL GLAND

这是一个基于两组初步数据的竞争性提案。 在第一个PI已经表明，培养的肾上腺皮质细胞释放 向上清液中加入一种或多种Na，K泵的哇巴因样抑制剂。 在第二个实验中，PI开发了一种高效液相色谱辅助定量反相 定量mRNA的转录-聚合酶链反应（QRTPCR）方法 少量组织中的水平。 基于这些，他提出了两个 目标。 目标1是利用几种现代分析技术， 表征肾上腺Y1释放的Na，K泵抑制剂 细胞 这将通过初步的HPLC纯化相对 大量的上清液培养基，随后是串联质量 如果材料达到足够的纯度，则通过高纯度， 分辨率NMR。 高纯度材料的选定化学改性 也可用于帮助鉴定上述抑制剂。 目的2是通过上述QRTPCR定量mRNA水平 方法在大鼠肾单位的特定节段上， 饮食钠摄入量。

This a competitive proposal that is based on two sets of preliminary data. In the first the PI has shown that cultured adrenocortical cells release into the supernatant one or more ouabain-like inhibitors of the Na, K pump. In the second, the PI has developed an HPLC-assisted quantitive reverse transcription-polymerase chain reaction (QRTPCR) method to quantitate mRNA levels in small amounts of tissue. Based on these, he is proposing two aims. Aim 1 is to utilize several modern analytical techniques to characterize the inhibitor (s) of the Na, K pump released by adrenal Y1 cells. This will done via preliminary HPLC purification of relatively large amounts of supernatant medium which will be followed by tandem mass spectrometry and if the material reaches sufficient purity, by high resolution NMR. Selected chemical modifications of high purity material may also be used to aid in identification of the aforementioned inhibitor. Aim 2 is to quantitate the levels of mRNA via the aforementioned QRTPCR method in defined segments of the rat nephron under different levels of dietary sodium intake.