MOLECULAR BIOLOGY OF CYCLIC HEMATOPOIESIS

长期的目标是确定的分子病理生理学， 灰色牧羊犬的循环造血（CH），并评估在 重组造血生长因子在治疗中的体内应用 周期性造血 探讨周期性脑梗死的病理生理学 我们将描述一种最近发现的 磷酸肌醇第二信使产生的生化缺陷， 调查生产中可能存在的缺陷或 CH犬的造血生长因子。 具体目标和 方法是： 1)探讨磷脂酶C/蛋白的生化基础 激酶C介导的蛋白磷酸化缺陷， 甘油二酯和磷酸肌醇生产，蛋白激酶 C激活和膜转位并比较G蛋白 正常和CH犬中磷脂酶C的调节。 2)表征调节周期性G-CSF产生的机制， 确定是否存在造血生长的周期性产生 因子IL-3、GM-CSF、M-CSF和单核因子IL-1和肝细胞 用cDNA或寡核苷酸探针定量测定活化因子 mRNA的产生。 骨髓和因子依赖性细胞系将 用于定量生长因子和单核因子的水平， 来自骨髓培养物的血清和条件培养基。 3)比较正常和CH犬骨髓细胞的反应 与重组生长因子的体外CFU测定。 4)重组造血生长因子对CH犬的给药 研究对造血的体内作用，并确定其 调节造血和刺激生长的潜力 休眠的骨髓细胞 复杂的体内控制机制调节造血， 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。 造血干细胞的潜在治疗用途 生长因子和生长因子组合在刺激 静止骨髓细胞在体内的生长将被系统地 研究了 这些研究将提供新的信息， 造血干细胞调控与血细胞疾病 生产

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.