THE MOLECULAR BIOLOGY OF CYCLIC HEMATOPOIESIS

长期的目标是定义脑血管疾病的分子病理生理学。 灰色牧羊犬的循环造血功能及其临床意义 重组造血生长因子在体内治疗中的应用 周期性出血。探讨环磷酰胺的病理生理学机制 出血，我们将描述一个最近发现的 肌醇磷脂第二信使生产过程中的生化缺陷 调查生产或行动中可能存在的缺陷 脑出血犬的造血生长因子。具体目标和 方法包括： 1)探讨磷脂酶C/蛋白质的生化基础 蛋白激酶C介导的蛋白磷酸化缺陷(S) 二酰甘油和肌醇磷酸生产，蛋白激酶 C活化与膜转位及G蛋白的比较 正常和充血性心力衰竭犬磷脂酶C的调节。 2)表征调节循环G-CSF产生的机制和 判断是否存在造血细胞生长的循环产生 IL-3、GM-CSF、M-CSF与单因子IL-1、肝细胞 用cDNAs或寡核苷酸探针检测激活因子 信使核糖核酸的产生。骨髓和因子依赖细胞系将 可用于定量测定体内生长因子和单核细胞因子水平 骨髓培养的血清和条件培养液。 3)比较正常和CH犬骨髓细胞的反应 用体外CFU法检测重组生长因子。 4)给CH犬应用重组造血生长因子 研究体内对造血的作用并确定其在体内的作用 调节造血和刺激生长的潜力 静息的骨髓细胞。 复杂的体内调节造血的控制机制， 造血生长因子的产生和激活 造血细胞的神经内分泌轴将被表征 在这些研究中。造血剂的潜在治疗用途 生长因子和生长因子组合在刺激中的作用 体内静止的骨髓细胞的生长将被系统地 调查过了。这些研究将提供有关 造血干细胞调控与血细胞紊乱 制作。

The long-term goals are to define the molecular pathophysiology of cyclic hematopoiesis (CH) in gray collie dogs and evaluate the in vivo use of recombinant hematipoietic growth factors in treating cyclic hematipoiesis. To investigate the pathophysiology of cyclic hematipoiesis we will characterize a recently identified biochemical defect in phosphoinositide 2nd messenger production and investigate possible defects in production or actions of hematopoietic growth factors in CH dogs. The specific aims and methods are: 1) Investigate the biochemical basis for phospholipase C/protein kinase C-mediated protein phosphorylation defect(s) by measuring diacylglycerol and inositol phosphate production, protein kinase C activation and membrane translocation and compare G-protein regulation of phospholipase C in normal and CH dogs. 2) Characterize mechanisms regulating cyclic G-CSF production and determine if there is cyclic production of the hematopoietic growth factors IL-3, GM-CSF, M-CSF and the monokines IL-1 and hepatocyte activating factor with cDNA or oligonucleotide probes to quantitate mRNA production. Bone marrow and factor dependent cell lines will be used to quantitate levels of growth factors and monokines in serum and conditioned media from bone marrow cultures. 3) Compare the response of normal and CH dog bone marrow cells to recombinant growth factors with an in vitro CFU assay. 4) Administer recombinant hematipoietic growth factors to CH dogs to investigate in vivo actions on hematopoiesis and determine their potential for modulating hematopoiesis and stimulating growth of resting marrow cells. The complex in vivo control mechanisms regulating hematopoiesis, hematopoietic growth factor production and activation of the neuroendocrine axis by hematopoietic cells will be characterized in these studies. The potential therapeutic use of hematopoietic growth factors and growth factors combinations in stimulating growth of quiescent marrow cells in vivo will be systematically investigated. These studies will provide new information on hematopoietic stem cell regulation and disorders of blood cell production.