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首页> 外文学位 >Development of a novel fouling deterrence strategy by understanding the effect of noradrenaline on the cells of Eastern Oyster, Crassostrea virginica and cypris larvae of the Striped Barnacle Balanus amphitrite.
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Development of a novel fouling deterrence strategy by understanding the effect of noradrenaline on the cells of Eastern Oyster, Crassostrea virginica and cypris larvae of the Striped Barnacle Balanus amphitrite.

机译：通过了解去甲肾上腺素对条纹藤壶Balampus amphitrite的东部牡蛎，Crassostrea virginica和Cypris幼虫细胞的作用，开发一种新型的结垢威慑策略。

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摘要

The focus of this study was to understand and evaluate the effect of noradrenaline (NA) on two prominent fouling marine invertebrates, the Eastern Oyster, Crassostrea virginica and the Striped Barnacle, Balanus amphitrite for the purpose of developing a novel fouling deterrence strategy.; To understand the effect of NA at a cellular level the immune cells (hemocytes) of C. virginica were chosen as representative molluscan cells. Upon stimulation with 10 micro-molar of noradrenaline ∼4% hemocytes of C. virginica labeled positive for the beta-adrenergic receptor (beta-AR). Upon NA stimulation the beta-AR positive cells formed cell-cell synapses and apoptosis was detected in ∼50% of the hemocyte population.; NA induced apoptosis was followed from the earliest onset to the terminal stages, from the release of cytochrome-c to the DNA degradation. Electric cell substrate impedance measurements suggested that NA stimulation induced cytoskeletal changes in the hemocytes. SDS PAGE and Western analysis corroborated the presence of beta-AR on NA stimulated hemocytes. Mass spectral analysis of the receptor protein revealed that the putative hemocyte beta-AR has a 30% sequence identity to the human beta-AR.; Effect of NA on the settling behavior of B. amphitrite cypris larvae was evaluated by challenging the larvae with micromolar concentrations of NA ranging from 30-100 micro-molar. The noted searching behavior of cypris larvae was lost after NA challenge and the larvae failed to cement to the substratum. ∼70% of the NA treated larvae failed to settle and cement. NA treatment caused a considerable delay in cyprid-adult metamorphosis. Metamorphosed juvenile barnacles were observed 96 hours following NA treatment, whereas the control cyprids metamorphosed within 24-48 hours. In the cyprids-adult metamorphosis remnants of the cyprid stage were observed up to 48 hours of development. Metamorphosed juvenile barnacles appeared normal from the presence of growth increments and beating of cirri. NA challenge seemed to promote metamorphosis to juveniles forgoing the settling behavior and cementation to the substratum.; To assess the applicability of NA as a fouling deterrent, NA molecules were covalently conjugated to HEMA (2-hydroxyethyl methacrylate) and MAA (methacrylic acid) polymer surfaces. Ability of covalently conjugated NA molecules to activate their target adrenoreceptors was assessed by understanding the effect of NA-conjugated polymer surfaces on hemocytes of C. virginica.; NA conjugated polymer surfaces induced apoptosis in the hemocytes of C. virginica. Annexin-V assay confirmed the initiation of apoptosis.; Cytoskeletal structure of the hemocytes adhering to NA-conjugated polymer surfaces displayed pronounced degradation. Apoptotic blebbing of plasma membrane was also observed using scanning electron microscopy. Control polymer surfaces failed to exert any deleterious effects on the hemocytes.; Based on the results obtained in this study a novel fouling deterrence strategy is discussed.
机译：这项研究的重点是了解和评估去甲肾上腺素（NA）对两种主要的结垢海洋无脊椎动物，即东部牡蛎Crassostrea virginica和条纹藤壶Balanus amphitrite的作用，目的是开发一种新颖的结垢威慑策略。为了了解NA在细胞水平上的作用，选择了维吉尼亚衣原体的免疫细胞（血细胞）作为代表性的软体动物细胞。用10微摩尔的去甲肾上腺素刺激后，约4％的维尔纽斯血细胞标记为β-肾上腺素能受体（β-AR）阳性。 NA刺激后，β-AR阳性细胞形成细胞-细胞突触，在约50％的血细胞中检测到凋亡。从最初到结束，从细胞色素-c的释放到DNA降解，都遵循NA诱导的凋亡。电细胞底物阻抗测量表明，NA刺激可诱导血细胞中的细胞骨架变化。 SDS PAGE和Western分析证实了NA刺激的血细胞上存在β-AR。受体蛋白的质谱分析表明，假定的血细胞β-AR与人β-AR具有30％的序列同一性。通过用30-100微摩尔浓度的NA的微摩尔浓度挑战幼虫，评估NA对B.amphitrite cypris cypris幼虫的沉降行为的影响。 NA攻击后，失去了塞浦路斯幼虫的明显搜寻行为，并且该幼虫无法固定在基质上。经NA处理的幼虫中约有70％无法定居并固结。 NA治疗导致塞浦路斯成人变态的显着延迟。 NA处理96小时后观察到变态的幼年藤壶，而对照鲤在24-48小时内变态。在塞浦路斯成人变态阶段，在发育至48小时之前都观察到了塞浦路斯阶段的残留物。变态的幼体藤壶从生长的增加和卷毛的跳动看来是正常的。 NA挑战似乎促进了少年的变态，放弃了其沉降行为并使其固定在基底上。为了评估NA作为防污剂的适用性，将NA分子共价缀合到HEMA（甲基丙烯酸2-羟乙酯）和MAA（甲基丙烯酸）聚合物表面。通过了解NA共轭聚合物表面对维吉尼亚衣原体血细胞的影响，评估了共价共轭NA分子激活其靶肾上腺受体的能力。 NA缀合的聚合物表面诱导了维吉尼亚衣藻的血细胞凋亡。 Annexin-V测定证实了细胞凋亡的开始。黏附于NA共轭聚合物表面的血细胞的细胞骨架结构显示出明显的降解。使用扫描电子显微镜还观察到质膜的细胞凋亡。对照聚合物表面不能对血细胞产生任何有害作用。基于本研究获得的结果，讨论了一种新颖的防污策略。

著录项

  • 作者

    Gohad, Neeraj Vijay.;

  • 作者单位

    Clemson University.$bBiological Sciences.;

  • 授予单位 Clemson University.$bBiological Sciences.;
  • 学科 Biology Cell.; Biology Zoology.; Environmental Sciences.; Agriculture Fisheries and Aquaculture.
  • 学位 Ph.D.
  • 年度 2008
  • 页码 121 p.
  • 总页数 121
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 细胞生物学;动物学;环境科学基础理论;水产、渔业;
  • 关键词

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