HEMATOPOIETIC STEM CELL DIFFERENTIATION IN HUMAN BLOOD

这里提出的研究的长期目标是确定信号 生长因子刺激的转导机制 造血细胞的增殖和分化。这一行动的主要目标是 该项目旨在了解促红细胞生成素 促红细胞生成素(EPO)调节红系分化过程中的离子通道 确定钙内流在红细胞生成中的功能作用。 抽血和骨髓抽吸术将在 关于健康志愿者的儿童权利公约。以下是具体目标 处理：具体目标1：确定信号传递机制 促红细胞生成素通过其调节钙通道。(A)我们将在这里 确定将JAK2连接到钙通道的EPO信号通路 激活。STAT、RAS或IRS-2/PI 3激酶通路的参与 将使用单个BFU-E衍生的微量注射进行检查 红细胞和定量荧光显微镜数码联用 视频成像。(B)我们将确定促红细胞生成素的结构域 促红细胞生成素调节钙通道开放所需的受体。 具体目标2：确定[Cai]在调节 转录因子在红细胞生成中的激活。的功能 核转录因子-kB和碱性成纤维细胞生长因子与c-jun氨基末端激酶 (JNK)受钙的调节。我们将确定是波幅还是 红系细胞内钙反应持续时间对核因子-kB的影响 转录因子激活，或钙/钙调蛋白水平 影响bHLH蛋白的DNA结合，特别是SCL。我们还将 测定[CaI]对JNLK激活的影响 这一途径中的一种激活剂。

The long term goal of studies proposed here is to determine the signal transduction mechanisms through which growth factors stimulate hematopoietic proliferation and differentiation. The major goal of this project is to understand the mechanisms through which erythropoietin (Epo) regulates ion channels during erythroid differentiation and to determine the functional role of calcium influx in erythropoiesis. Blood will be drawn and bone marrow aspirates will be performed in the CRC on healthy volunteers. The following specific aims will be addressed: Specific Aim 1: Identification of the signaling mechanism through which Epo regulates calcium channels. (A) Here we will determine the Epo signaling pathways which link Jak2 to calcium channel activation. Involvement of STAT, Ras, or the IRS-2/PI 3 kinase pathways will be examined using microinjection of single BFU-E derived erythroblasts and quantitative fluorescence microscopy coupled digital video imaging. (B) We will determine the domains of erythropoietin receptor required for Epo modulation of calcium channel opening. Specific Aim 2:Determination of the role of [Cai] in regulation of transcription factor activation in erythropoiesis. The function of the NF-kB and bHLH transcription factors and the c-Jun N-terminal kinases (JNK) are modulated by calcium. We will determine if the amplitude or duration of the Ca++ response in erythroid cells affects NF-kB transcription factor activation, or if calcium/calmodulin levels influence DNA binding of bHLH proteins, particularly SCL. We will also determine the effect of [Cai] on JNLK activation on the role of PI 3- kinase in this pathway.