AGE RELATED RESPONSE OF SALIVARY GLANDS TO ISOPROTERENOL

唾液腺产生的分泌物可以沐浴和保护口腔。 胱抑素S，一种天然存在的半胱氨酸蛋白酶抑制物，由 颌下腺，被认为起着重要的保护作用。直到 近年来，对半胱氨酸氨基转移酶S基因的调控知之甚少。 尽管这些蛋白在口腔病理生理学中起着作用，但它们仍能在口腔中表达。 我们的长期目标是确定调控胱抑素的机制 基因。拟议研究的目标是确定功能 大鼠胱抑素S基因的调控元件，与人类高度同源 半胱氨酸氨基转移酶S基因，以及调控其基因的DNA结合蛋白 功能。具体目的是：1)检查荷尔蒙、神经元和 大鼠半胱氨酸氨基转移酶S基因的发育调控，2)测定 胱抑素S基因中的功能顺式作用DNA调控元件，以及 3)鉴定、分离和鉴定反式作用DNA结合蛋白 调控大鼠胱抑素S基因及其编码基因。我们会 生后大鼠颌下腺组织中胱抑素S基因的表达 去势和激素后雄性和雌性大鼠的发育期 替代，以及神经支配受阻的腺体。转染腺病毒载体 胱抑素S基因5‘端片段的构建 将猫报告基因导入适当的受体细胞将使我们能够 确定我们已经确定的哪些潜在的调控序列 都是有功能的。凝胶漂移和DNase I保护分析将允许我们 鉴定调控该基因的DNA结合蛋白。净化这些东西 蛋白质，获得其氨基酸序列并合成 与所述氨基酸序列相对应的寡核苷酸并生成 这些DNA结合蛋白的抗体将使克隆和 确定编码这些调节蛋白的基因的特征。了解以下内容 半胱氨酸氨基转移酶基因的表达调控可能导致临床 牙周病患者口腔功能障碍的治疗 疾病、干燥综合征和艾滋病毒疾病。

Salivary glands produce secretions that bathe and protect the oral cavity. Cystatin S, a naturally occurring cysteine proteinase inhibitor secreted by submandibular glands, is believed to play a major protective role. Until recently, nothing was known about the regulation of cystatin, S gene expression despite the role these proteins play in oral pathophysiology. Our long-range goal is to determine the mechanisms regulating cystatin genes. The goal of the proposed research is to identify the functional regulatory elements of the rat cystatin S gene, highly homologous to human cystatin S genes, and the DNA binding proteins that modulate their function. The Specific aims are to: 1) examine the hormonal, neuronal, and developmental regulation of the rat cystatin S gene, 2) determine the functional cis-acting DNA regulatory elements in the cystatin S gene, and 3) identify, isolate and characterize the trans-acting DNA binding proteins regulating the rat cystatin S gene and the genes encoding them. We will examine cystatin S mRNA levels in the submandibular glands of postnatally developing males, of male and female rats after castration and hormone replacement, and in glands whose innervation was blocked. Transfection of segments of the 5' region of the cystatin S gene in constructs containing the CAT reporter gene into appropriate recipient cells will allow us to determine which of the potential regulatory sequences we have identified are functional. Gel shift and DNase I protection assays will permit us to identify the DNA binding proteins regulating this gene. Purifying these proteins, obtaining their amino acid sequence and synthesizing oligonucleotides corresponding to the amino acid sequence and generating antibodies to these DNA binding proteins will make it possible to clone and characterize the genes encoding these regulatory proteins. Knowledge of how expression of cystatin genes is regulated may lead to clinical treatments for patients who are orally compromised because of periodontal disease, Sjogren's syndrome, and HIV disease.