PURINOCEPTOR CONTROL OF SURFACTANT SECRETION

这项建议是基于我们最近确定并表达了一种 肺P2-嘌呤受体的特异性cDNA.这项提案代表了 首次描述在任何系统中克隆P2-嘌呤受体。我们会 现在验证一下肺中的P2-嘌呤受体调节 正常肺功能所需的表面活性物质磷脂的分泌 这些受体的表达和成熟是暂时的和 在围产期受荷尔蒙调节。《公约》的具体目标 目前的建议是完成cDNA的核苷酸序列 编码人和大鼠II型细胞P2-嘌呤受体。永久性细胞 将产生表达II型细胞P2-嘌呤受体的细胞系 用于研究受体的荷尔蒙调节。反义cRNA 将利用探针进行原位杂交来确定细胞- II型细胞P2-嘌呤受体的特异性和个体原性表达 阿龙。最后，反义寡核苷酸和P2-嘌呤受体 将产生抗体并用来证明这种关系 P2-嘌呤受体表达对表面活性物质磷脂的调节作用 分泌物。这些研究试图第一次定义 参与P2调节的P2-嘌呤受体和对照 嘌呤受体在肺中的表达。对结构的进一步了解 而肺中P2-嘌呤受体的个体发生调节将是有价值的 在开发肺部疾病治疗的新策略方面。

This proposal is based on our recent identification and expression of a cDNA specific for a lung P2-purinoceptor. This proposal represents the first description of cloning of a P2-purinoceptor in any system. We will now test the hypothesis that P2-purinoceptors in the lung regulate secretion of surfactant phospholipid necessary for normal lung function and that expression and maturation of these receptors is temporally and hormonally regulated in the perinatal period. The specific aims of the present proposal will be to complete the nucleotide sequence of the cDNAs encoding human and rat Type II cell P2-purinoceptors. Permanent cell lines expressing the Type II cell P2-purinoceptor will be generated and utilized to study hormonal regulation of the receptor. Antisense cRNA probes will be utilized for in situ hybridization to determine cell- specific and ontogenic expression of the Type II cell P2-purinoceptor in lung. Finally, antisense oligodeoxynucleotides and P2-purinoceptor antibodies will be generated and utilized to demonstrate the relationship of P2-purinoceptor expression to regulation of surfactant phospholipid secretion. These studies seek to define for the first time the nature of P2-purinoceptors and controls involved in regulation of P2- purinoceptor expression in the lung. Further knowledge of the structure and ontogenic regulation of P2-purinoceptors in the lung will be valuable in developing new strategies for therapy of lung diseases.