AGE RELATED RESPONSE OF SALIVARY GLANDS TO ISOPROTERENOL

唾液腺产生的分泌物可以清洗和保护口腔。 胱抑素S，一种天然存在的半胱氨酸蛋白酶抑制剂，由 下颌下腺被认为起着主要的保护作用。 直到 目前对cystatin，S基因的调控尚不清楚 尽管这些蛋白质在口腔病理生理学中起作用，但它们的表达仍然存在。 我们的长期目标是确定半胱氨酸蛋白酶抑制剂的调节机制， 基因. 这项研究的目的是确定 大鼠半胱氨酸蛋白酶抑制剂S基因的调控元件，与人类高度同源 半胱氨酸蛋白酶抑制剂S基因，以及调节它们的DNA结合蛋白。 功能 具体目标是：1）检查激素，神经元， 大鼠胱抑素S基因的发育调控，2）确定 胱抑素S基因中的功能性顺式作用DNA调控元件，和 3)鉴定、分离和表征反式作用DNA结合蛋白 调控大鼠胱抑素S基因及其编码基因。 我们将 检测生后大鼠下颌下腺中胱抑素S mRNA的表达 去势和激素后雄性和雌性大鼠的发育中雄性 替代，并在腺体的神经支配被阻断。 转染 构建体中的半胱氨酸蛋白酶抑制剂S基因的5'区的片段，所述构建体含有 将CAT报告基因导入合适的受体细胞， 确定我们已经识别出的潜在调控序列 是功能性的。 凝胶迁移和DNA酶I保护试验将使我们能够 鉴定调节该基因的DNA结合蛋白。 净化这些 蛋白质，获得其氨基酸序列并合成 所述寡核苷酸对应于所述氨基酸序列并产生 这些DNA结合蛋白的抗体将使克隆和 表征编码这些调节蛋白的基因。 知识 半胱氨酸蛋白酶抑制剂基因的表达是如何调节的， 牙周病患者的治疗方法 疾病、干燥综合征和HIV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.