SALIVARY GLAND SECRETORY MECHANISMS DURING NORMAL AND ALTERED FUNCTIONAL STATES

正常和改变功能状态期间的唾液腺分泌机制

• 批准号: 3753528
• 负责人: B J BAUM
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3753528
• 关键词: Adenoviridae; adeno associated virus group; biological signal transduction; genetic manipulation; human subject; membrane channels; membrane permeability; protein signal sequence; salivary glands; secretion; transfection; transfection /expression vector; water flow

The health of the oral cavity is maintained by salivary secretions. The principal function of salivary glands is to produce these complex fluids. We utilize in vitro dispersed cells of salivary glands, in vivo cannulated glands, and cultured salivary cell lines as laboratory models to understand mechanisms controlling saliva formation. We have focused most of our studies on neurotransmitter regulation of secretory events. During this reporting period the primary focus of signaling studies continues to be muscarinic receptors (mAChRs) in rat parotid gland acinar cells. In parotid cells, stimulation of mAChRs results in the generation of inositol phosphates via the activation of phospholipase C. Subsequently this response leads to the elevation of cytosolic Ca2+ levels and fluid secretion. We have continued to characterize mAChRs in intact rat parotid cells using the binding of a subtype non-selective antagonist (NMS, N-methylscopolamine). We have determined that a moderate population (approximately 30-40%) of spare receptors exist for inositol trisphosphate formation. We have continued in vivo studies of mAChRs in exocrine glands using iodinated QNB (quinuclidinyl benzilate) enantiomers and phamacokinetic analyses. These experiments have led to the development of a clinical research protocol to examine mAChRs in normal human volunteers. To understand how salivary glands transport water we have begun studies on a putative water channel, CHIP28. We have isolated a cDNA encoding a CHIP28-like protein from a rat parotid library and examined its cellular distribution in this gland by in situ hybridization. We have also initiated efforts to transfer foreign genes into rat salivary glands in vivo using replication deficient recombinant adenovirus (Ad) vectors (e.g. containing genes encoding E. Coli beta-galactosidase, beta gal; and human alpha 1 antitrypsin, alpha1AT). Two days after retrograde duct instillation of Ad-beta gal striking expression is seen in acinar and ductal cells of all major salivary glands. Transfer of the alpha1AT gene results in secretion of this protein in gland saliva for 4-10 days.

口腔的健康是由唾液分泌物维持的。 唾液腺的主要功能是产生这些复合物 流体。 我们利用唾液腺的体外分散细胞， 体内插管的腺体和培养的唾液细胞系作为实验室 了解控制唾液形成的机制的模型。 我们有 将大多数研究集中在分泌的神经递质调节上 事件。 在此报告期间，信号的主要重点 研究继续是大鼠腮腺中的毒蕈碱受体（MACHR） 腺腺泡细胞。 在腮腺细胞中，刺激MACHRS结果 通过激活产生肌醇磷酸盐 磷脂酶C.随后这种反应导致 胞质Ca2+水平和流体分泌。 我们继续 使用结合的结合来表征完整的大鼠腮腺细胞中的MACHR 亚型非选择性拮抗剂（NMS，N-甲基苏氨基胺）。 我们有 确定备用中等人口（约30-40％） 存在于肌醇三磷酸形成的受体。 我们有 继续使用碘化 QNB（奎尼核苷酸苯甲酸酯）对映异构体和phamacokinetic分析。 这些实验导致了临床研究的发展 在正常人类志愿者中检查MACHR的方案。 理解 唾液腺如何运输我们已经开始研究推定的水 水通道，chip28。 我们已经隔离了编码chip28样的cDNA 大鼠腮腺库的蛋白质并检查了其细胞 通过原位杂交在该腺中的分布。 我们也有 开始将外源基因转移到大鼠唾液腺的努力 使用复制不足的重组腺病毒（AD）向量的体内体内 （例如，包含编码大肠杆菌β-半乳糖苷酶的基因，βgal; 和人α1抗胰蛋白酶，α1AT）。 逆行两天后 在腺泡中可以看到ad-beta gal醒目表达的管道滴注 和所有主要唾液腺的导管细胞。 转移 α1AT基因导致该蛋白在腺唾液中的分泌 4-10天。