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Abstract
Protocol
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Immunologie et infection

唾液，唾液腺，淋巴收集肩突硬蜱蜱

Published: February 21, 2012
doi:
10.3791/3894
, , , , ,
1Microbiology and Pathogenesis Activity, Bacterial Diseases Branch, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases,Centers for Disease Control and Prevention, 2Tick-Borne Diseases Activity, Bacterial Diseases Branch, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases,Centers for Disease Control and Prevention

Summary

蜱血淋巴感染，唾液腺和唾液的收集是非常重要的研究蜱传病原体引起疾病。在这个协议中，我们演示了如何收集喂血淋巴和唾液腺<em>肩突硬蜱</em>若虫。我们还表明，从女性的唾液收集<em>一。肩胛</em>成年人。

Abstract

蜱遍布世界各地，并与许多蜱传疾病的折磨人类。蜱是莱姆病和蜱传回归热（ 伯氏菌），洛矶山斑疹热（ 立克次氏体 ），埃立克体病（ 查菲埃立克体和E.球菌 ），无形体病（ 嗜吞噬细胞无形体 ），脑炎（蜱的病原体引起的载体森林脑炎病毒），巴贝斯虫病（ 巴贝斯虫属），科罗拉多蜱热（Colti病毒），兔热病（ 弗朗西斯tularensis）1-8。要正确传输到主机，这些传染性病原体的差异调节基因表达，与蜱蛋白质交互，并通过蜱3,9-13迁移。例如，莱姆病代理， 莱姆病 ，蜱的地方性周期14,15盛宴和饥荒阶段适应，通过基因差异表达。此外，作为硬蜱蜱消耗吸血伯从肠到的hemocoel，他们前往唾液腺，并传送到主机与被驱逐的唾液9,16-19复制和迁移。

由于蜱送入主机通常响应与一个强大的止血和先天免疫反应11,13,20-22。尽管这些宿主反应， 一肩胛可以养活了好几天，因为蜱唾液中含有的免疫调节的蛋白质，溶解剂，抗凝血剂，并fibrinolysins帮助蜱喂养3,11,20,21,23。由蜱唾液或唾液腺提取物（SGE）具有免疫调节活动方便，众多的蜱传病原体3,20,24-27的传输，扩散和传播。为了进一步了解如何蜱传播的传染性病原体引起的疾病，它是必不可少的积极剖析饲养蜱和收集蜱唾液。这个视频协议演示的夹层技术从积极喂养一血淋巴的收集和唾液腺的去除肩胛若虫后，48和72小时后鼠标的位置。我们还表明，从一成年女性唾液收集肩胛蜱。

Protocol

1。幻灯片准备的淋巴集合 （电影1） 动物和外用3％过氧化氢的地方，轻轻地从积极喂养蜱5分钟，然后在70％乙醇10分钟，到表面消毒。 与PAP笔硅烷涂层显微镜幻灯片上绘制一个圆，并放置在PAP笔圈的刻度。 硅烷涂层幻灯片用最好的淋巴坚持显微镜幻灯片。 在解剖显微镜下（1X目标，10X目镜，放大倍率3.5X）蜱。 用钳子轻轻地往下推?…

Discussion

蜱血淋巴，唾液腺和唾液的收集是非常重要的研究蜱传病原体传播，流行，传播，扩散，并在双方的滴答和主机6,11-13,20,23,29的持久性。有几种方法剖析蜱30,31的 。然而，当收集唾液腺关键是正确剖析蜱唾液腺不破裂，或在蜱的遗体丢失。一旦被删除要洗几次，清除肠污染，然后可以固定获得唾液腺提取物（SGE）染色或PBS地面幻灯片上，他们需要从蜱唾液腺。上海黄金交易所是更?…

Divulgations

The authors have nothing to disclose.

Acknowledgements

作者要感谢病媒传播疾病的动物资源科司，特别是安德烈彼得森，丽莎马苏迪，贝尔纳奥布莱恩，约翰·利德尔的老鼠和兔子的关心和维护。我们也想感谢艾米·乌尔曼，特里萨·拉塞尔，和Barbara J.约翰逊朝着这个手稿的贡献。最后，我们想承认艾丽萨埃克特通信在疾病预防控制中心副主任办公室指导拍摄这个手稿与所有相关的合法性产生的插图和朱迪·拉维尔。

Materials

Reagent Company Catalogue Number
Hydrogen peroxide Fisher H312-500
Ethanol Acros 61509-5000
PBS Boston Bioproducts BM-2205
Dumont Fine forceps (3C) Fisher NC9906085
Silane treated microscope slides Bioworld 42763007-1
Pap pen Bioworld 21750008-1
Super frost plus microscope slides Fisher 12-550-18
Pilocarpine Sigma P6503-5G
Protease inhibitor cocktail Sigma P2714
#11 disposable scalpel Feather 2975#11
Nontoxic modeling clay Fisher S17307
Capillary tubes Chase scientific Glass, inc 40A502
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Tags

SalivaSalivary GlandHemolymph CollectionIxodes Scapularis TicksTick-borne IllnessesLyme DiseaseTick-borne Relapsing FeverRocky Mountain Spotted FeverEhrlichiosisAnaplasmosisEncephalitisBabesiosisColorado Tick FeverTularemiaBorrelia Spp.Rickettsia RickettsiiEhrlichia ChaffeensisE. EquiAnaplasma PhagocytophilumTick-borne Encephalitis VirusBabesia Spp.ColtivirusFrancisella TularensisGene Expression RegulationTick Proteins InteractionTick MigrationBorrelia BurgdorferiEnzootic CycleHemostatic ResponseInnate Immune ResponseTick Saliva Proteins
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Patton, T. G., Dietrich, G., Brandt, K., Dolan, M. C., Piesman, J., Gilmore Jr., R. D. Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks. J. Vis. Exp. (60), e3894, doi:10.3791/3894 (2012).
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