CONTROL OF CONJUNCTIVAL GOBLET CELL MUCOUS PRODUCTION

结膜杯状细胞粘液产生的控制

• 批准号: 2711038
• 负责人: Darlene A Dartt
• 金额: $ 26.98万
• 依托单位: SCHEPENS EYE RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-05-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2711038
• 关键词: autoradiography; confocal scanning microscopy; conjunctiva; efferent nerve; enzyme linked immunosorbent assay; eye disorder; goblet cells; immunocytochemistry; immunoelectron microscopy; immunofluorescence technique; laboratory rat; mucus; neuroregulation; neurotransmitter agonist; neurotransmitter antagonist; neurotransmitter receptor; parasympathetic nervous system; prostaglandin E; second messengers; secretion; substance P; sympathetic nervous system

The mucous layer constitutes a physical and chemical barrier that protects the ocular surface from bacterial and environmental insults. One of the most important sources of the mucous layer is conjunctival goblet cells. Goblet cell mucous secretion can be stimulated by a neural stimulus to the cornea via parasympathetic and sympathetic nerves near goblet cells. This suggests that goblet cells are able to respond rapidly to external challenges to the ocular surface by secreting mucus to protect the eye surface. Under normal conditions, minor stimuli to the ocular surface would ensure a constant low level secretion. A more significant stimulus, such as ocular trauma, would cause a greater protective secretory response. A mucin-deficiency disease such as anesthetic cornea, vitamin A deficiency, late keratoconjunctivitis sicca, Sjogrens syndrome, thermal and alkali burns would occur with a breakdown of reflex secretion. Breakdown could occur because of a dysfunction of the stimulating neural pathways, leading to a decreased mucous secretion, loss of the protective coat and deterioration of the ocular surface. In diseases of mucous overproduction, a constant irritative stimulus to the ocular surface would increase neural stimulation, and thus increase goblet cell secretion. These diseases include vernal conjunctivitis, giant papillary conjunctivitis, early keratoconjunctivitis sicca, mucous fishing syndrome, and atopy. Thus, the overall goal of the project is to determine the components of the different neural pathways that stimulate mucous secretion by investigating which nerves, neurotransmitters, receptors, second messengers and cell types are involved. First, the role of sensory, parasympathetic, and sympathetic nerves in stimulating conjunctival goblet cell mucous secretion will be investigated. Second, the identity and relationship of nerve fibers to conjunctival goblet cells will be determined. Third, the role of the prostaglandin PGE2 in neural stimulation of goblet cell secretion will be explored. For in vivo secretion experiments, rat conjunctival goblet cell secretion will be stimulated by a corneal wound or by topical application of agonists. Goblet cell mucin, in buttons of conjunctiva, will be stained histochemically and the number of goblet cells per area counted. A decrease in this number indicates an increase in mucous secretion. For in vitro secretion experiments, conjunctival tissue will be removed and incubated with or without agonists. Mucous secretion will be measured by an enzyme-linked lectin assay using Helix pomatia agglutinin, which preferentially recognizes goblet cell secretory product. The neural innervation of goblet cells will be determined by confocal microscopy with immunofluorescence labelling and by immunoelectron microscopy. Receptors on goblet cells will be localized by immunohistochemistry or autoradiography. Knowledge of the normal mechanism of neural regulation of goblet cell mucous secretion will provide a scientific basis to develop therapeutic treatments for diseases of the mucous layer.

粘液层构成物理和化学屏障， 保护眼表面免受细菌和环境的侵害。 其中一个最重要的来源是粘膜层是结膜 杯状细胞 杯状细胞粘液分泌可以被神经刺激， 刺激角膜通过副交感神经和交感神经附近 杯状细胞 这表明杯状细胞能够对 通过分泌粘液迅速应对眼表的外部挑战 保护眼睛表面。 在正常情况下，轻微刺激 眼表面将确保恒定的低水平分泌。 一个更 严重刺激，如眼外伤，会引起更大的 保护性分泌反应 一种粘蛋白缺乏性疾病，如 麻醉角膜，维生素A缺乏，晚期干燥性角结膜炎， 干燥综合征，热和碱烧伤会发生与故障 反射分泌。 崩溃可能是因为功能障碍， 刺激神经通路，导致粘液分泌减少， 保护层的丧失和眼表的退化。 在 粘液分泌过多的疾病，对皮肤的持续刺激 眼表会增加神经刺激，从而增加 杯状细胞分泌 这些疾病包括春季结膜炎， 巨乳头性结膜炎，早期干燥性角结膜炎，粘液性 钓鱼综合症和特应性反应 因此，该项目的总体目标是 以确定不同神经通路的组成部分， 通过研究哪些神经刺激粘液分泌， 神经递质、受体、第二信使和细胞类型是 涉案 一、感觉、副交感、交感神经的作用 神经刺激结膜杯状细胞分泌粘液， 研究了 第二，神经纤维的身份和关系， 将测定结膜杯状细胞。 第三， 前列腺素PGE2在刺激神经杯状细胞分泌时会 被探索。 对于体内分泌实验，将大鼠结膜杯状细胞培养液（coconjunctival glucose）置于培养皿中。 细胞分泌将被角膜伤口或局部刺激， 激动剂的应用。 杯状细胞粘蛋白，在结膜钮扣中， 将进行组织化学染色，每个区域的杯状细胞数量 算 这个数字的减少表明粘液的增加， 分泌物 对于体外分泌实验，结膜组织将 取出并与或不与激动剂一起孵育。 粘液分泌物会 通过酶联凝集素测定法，使用Hessopomatia 凝集素，它优先识别杯状细胞分泌 产品 杯状细胞的神经支配将由 免疫荧光标记共聚焦显微镜， 免疫电子显微镜。 杯状细胞上的受体将定位于 免疫组织化学或放射自显影。 正常知识 杯状细胞粘液分泌的神经调节机制 为开发疾病治疗方法提供科学依据 的粘液层。