CARDIAC GLYCOSIDES FROM THE ADRENAL GLAND

这是一个基于两组初步数据的竞争性提案。 在第一个实验中，PI显示培养的肾上腺皮质细胞释放 进入上清液中的一个或多个哇巴因样的Na，K泵抑制剂。 在第二种情况下，PI开发了一种高效液相辅助的定量逆转 转录-聚合酶链式反应(QRTPCR)方法定量检测mRNA 在少量组织中的含量。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来 肾上腺Y1释放的Na，K泵抑制剂S的特征 细胞。这将通过相对高效液相色谱初步纯化来完成。 大量的上清液，随后将是串联质量 光谱和如果材料达到足够的纯度，通过高 解析核磁共振。高纯材料的选择性化学改性 也可用于辅助鉴定上述抑制剂。 目的2是通过上述QRT-PCR定量检测mRNAs的水平。 方法在不同水平的大鼠肾单位的不同节段 膳食钠摄入量。

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.