REPAIR OF HAIR BUNDLES IN SEA ANEMONES

海葵毛束的修复

基本信息

批准号：
2883021
负责人：
GLEN M WATSON
金额：
$ 10.79万
依托单位：
UNIVERSITY OF LOUISIANA AT LAFAYETTE
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-03-08 至 2001-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2883021
关键词：
Anthozoa SDS polyacrylamide gel electrophoresis chemical binding chemical structure function cilium hair immunocytochemistry interference microscopy ion exchange chromatography mechanoreceptors scanning transmission electron microscopy sensory receptors video microscopy wound healing

项目摘要

The proposed research constitutes the first effort to investigate the mechanism by which hair bundles on tentacles of sea anemones self-repair. Whereas certain avian species are known to respond to trauma to their hair bundles by replacing entire hair cells, no organism is yet known to repair existing hair bundles. Preliminary data detailed in this proposal suggest that repair is accomplished by proteins secreted by the anemone in response to trauma to its hair bundles. Research plans include fractionation of repair proteins (exclusion chromatography and ion- exchange chromatography). Fractions will be tested for biological activity (i.e., the ability to shorten the time course of recovery in test animals). Biologically active fractions will be subjected to SDS-PAGE gels. Specific, purified repair proteins will be biotinylated. Provided such proteins retain biological activity, they will be used in combination with avidin-fluorochromes or with avidin-colloidal gold to localize binding sites for repair proteins on the anemone at the light (confocal) and electron (field emission scanning) microscopic levels. Any repair proteins losing biological activity as a result of biotinylation will be used as antigens to generate polyclonal antibodies. The antibodies will be used in immunocytochemistry. Video-enhanced differential interference contrast (DIC) microscopy will be used to investigate the repair process on hair bundles of living specimens. Field emission scanning microscopy will be used to characterize the effects of repair proteins on linkages (including tip links) interconnecting stereocilia of the hair bundle. Long range plans for this project including testing the usefulness of anemone repair proteins as possible therapeutic agents for damaged hair bundles in vertebrates.

拟议的研究是调查这一问题的第一项努力海葵触手上的毛束自我修复的机制。众所周知，某些鸟类会对毛发创伤做出反应通过替换整个毛细胞来修复束，目前尚无有机体可以修复现有的发束。本提案中详述的初步数据表明修复是通过海葵分泌的蛋白质完成的对毛束创伤的反应。研究计划包括修复蛋白的分级分离（排除色谱法和离子色谱法）交换色谱）。馏分将进行生物学测试活动（即缩短测试恢复时间的能力）动物）。生物活性组分将进行 SDS-PAGE 凝胶。特定的、纯化的修复蛋白将被生物素化。假如此类蛋白质保留生物活性，它们将组合使用使用抗生物素蛋白-荧光染料或抗生物素蛋白-胶体金进行定位光下海葵上修复蛋白的结合位点（共聚焦）和电子（场发射扫描）微观水平。任何维修由于生物素化而失去生物活性的蛋白质将用作产生多克隆抗体的抗原。抗体会可用于免疫细胞化学。视频增强差分干扰对比（DIC）显微镜将用于研究修复过程在活体标本的发束上。场发射扫描显微镜将用于表征修复蛋白对连接的影响（包括尖端链接）互连毛束的静纤毛。该项目的长期计划包括测试海葵修复蛋白作为受损头发的可能治疗剂脊椎动物中的束。