Name: harvesting venom toxins from assassin bugs and other heteropteran insects
Uploaded: 2018-04-21T13:00:00
Description: Watch this Scientific Journal Video about Harvesting Venom Toxins from Assassin Bugs and Other Heteropteran Insects at JoVE.com

我们承认澳大利亚研究理事会的财政支持 (赠款 DP130103813 和 LP140100832 G.F.K., DECRA 奖学金 DE160101142 EABU), 澳大利亚国家健康 & 医学研究理事会 (主要研究金APP1044414 到 G.F.K.) 和昆士兰大学 (博士后奖学金 A.A.W.)。

该议定书符合昆士兰大学在教学和研究中负责照料和使用动物的政策 (4.20.11) 以及国家卫生和医学研究理事会的澳大利亚保护和使用动物为科学目的(8th编辑 2013)。
注意: 在处理刺客 bug 时, 注意不要被 envenomated。在处理毒液防御的物种时, 要注意保护眼睛。注意不要伤害实验动物。这包括监测压力的限制, 如橡胶带和确保喙没有打破。
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<ol>
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捕获刺客臭虫毒液最关键的一步是根据研究的目的选择合适的方法。三种收集 heteropteran 毒液的方法中, 每种都有优缺点, 这取决于下游的应用。
诱导臭虫从喙中排出毒液 (1-3 协议) 避免了腺组织对毒液的污染。此外, 这些方法是非致命的, 可以重复多次在一个 bug 的生活过程中。刺激通常提供最大数量的毒液, 并产生毒液具有强大的毒性对猎物昆虫根据几个研究5</s...

Heteropteran 毒液是完全生物活性物质1。例如, 血液喂养翅的毒液/唾液分泌物, 如接吻臭虫 (Triatominae) 和床虫 (Cimicidae) 通过扰乱止血2来促进喂养。这些毒液中的毒素靶向多个途径, 包括凝血、血小板聚集和血管收缩, 以及疼痛和瘙痒通路。大多数其他 heteropteran 物种的毒液都适于捕食, 而不是供血。他们的毒液导致瘫痪, 死亡和组织液化时, 注入无脊椎动物3,4。当注入脊椎动物时, 它们的毒液也可能产生剧烈的影响。例如, 从刺客 bug Holotrichius innesi中注入毒液会导致疼痛、肌肉麻痹和出血;由于呼吸麻痹5, envenomated 的老鼠很快就死了。
Transcriptomic 和蛋白质组学研究揭示了一些 heteropteran 毒液的蛋白质组成。捕食性天敌种类的毒液富含蛋白酶、其他酶, 以及未知结构和功能的多肽和蛋白质6, 7, 8.接吻臭虫毒液富含 triabin 蛋白家族, 其成员对凝血、血小板聚集和收缩29产生深刻影响。然而, 它不知道哪些毒素的基础上最具生物活性的毒液。例如, 接吻 bug Triatoma 疫病的毒液被报告为止痛药和抑制钠通道10, 但负责的组件仍有待阐明。同样, 还不知道刺客臭虫毒液的成分会导致麻痹或疼痛。确定对特定毒液生物活性负责的毒素, 以及表征新型毒液毒素的结构和功能的先决条件是获取毒液。
毒液已经从 heteropterans 获得了由刺激5,6,7,8,11,12,13, 挑衅防御响应4,8, 机械地压缩胸部12,14,15,16, 剖析毒液腺体8,17 、18、19、20、21、22和毒蕈乙酰胆碱受体的激动剂23的应用。判断任何方法的潜在优点和缺点是复杂的 heteropteran 毒液腺体的形态学, 它由一个主腺体与两个独立的流明, 前主腺体 (AMG) 和后主腺体 (普纳), 以及伴生的附属的腺体 (AG)。这些不同的腺体间隔产生不同的蛋白质分泌物, 这可能是专门用于不同的生物功能, 包括猎物捕获, 防御和额外口服消化8,17。在 peiratine 和 ectrichodiine 刺客臭虫, AMG 已经与猎物捕获和普纳的额外口服消化17。但是, 在 harpactorine bug Pristhesancus plagipennis中, 普纳专门用于捕获和消化, 而 AMG 则被假定为分泌防御毒液8。该 AG 已被描述为在刺客 bug8或作为一个主要的地方蛋白酶存储在巨型水臭虫23中的分泌功能很少。显然, 需要进一步的工作来澄清各个 heteropteran 子群之间各腺体间的功能, 并确定大多数毒液毒素的功能。在本报告中, 我们描述了从 heteropterans 到这个目标的收集毒液毒素的协议。

<table border="0" cellpadding="0" cellspacing="0" width="912">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Electostimulator</td>
			<td style="width:203px;">Grass Technologies</td>
			<td style="width:187px;">S48 Square Pulse Stimulator</td>
			<td style="width:307px;">Electrostimulator allowing pulsed electrostimulation</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Featherlight tweezers</td>
			<td style="width:203px;">Australian Entomological Supplies</td>
			<td style="width:187px;">E122B</td>
			<td>For handling live venomous insects</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Protease inhibitor cocktail</td>
			<td style="width:203px;">Sigma</td>
			<td style="width:187px;">4693124001</td>
			<td>For preventing autoproteolytic digestion of venom</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Dissection equipment</td>
			<td style="width:203px;">Australian Entomological Supplies</td>
			<td style="width:187px;">E152Micro</td>
			<td>For fine dissections</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Insect pins</td>
			<td style="width:203px;">Australian Entomological Supplies</td>
			<td style="width:187px;">E162</td>
			<td>For fine dissections</td>
		</tr>
	</tbody>
</table>

harvesting venom toxins from assassin bugs and other heteropteran insects

Heteropteran 昆虫, 如刺客臭虫 (Reduviidae) 和巨大的水虫 (Belostomatidae) 从一个共同的捕食性天敌和恶毒的祖先后裔, 和大多数现存的 heteropterans 保留这一营养战略。一些 heteropterans 已经转变为喂养脊椎动物的血液 (如接吻臭虫, Triatominae; 和床虫, Cimicidae), 而另一些已经恢复到植物喂养 (大多数 Pentatomomorpha)。然而, 除了接吻虫子用来促进血液喂养的唾液之外, heteropteran 毒液与蜘蛛、蝎子和蛇的毒液相比, 很少被知晓。
heteropteran 毒液毒素特征的一个障碍是毒液/唇腺的结构和功能, 它们既形态复杂, 又具有多种生物作用 (防御、猎物捕获和超口服消化)。在本文中, 我们描述了三种方法, 我们已经成功地用于收集 heteropteran 毒液。首先, 我们提出刺激作为一种方便的方法收集毒液, 这往往是致命的, 当注射到猎物的动物, 并省却污染的腺组织。第二, 我们表明, 对动物的温柔骚扰足以产生毒液挤压从喙和/或毒液随地吐痰的一些群体的 heteropterans。第三, 我们描述的方法, 以收集毒液毒素的麻醉动物的解剖, 以获得毒液腺体。这种方法与其他方法相辅相成, 因为它可能允许从刺激和骚扰无效的分类群中收集毒素。这些协议将使研究人员能够从 heteropteran 昆虫身上获得毒素, 用于结构功能表征和在医药和农业中的可能应用。

某些 heteropteran 物种 (如 harpactorine P plagipennis和 reduviine Platymeris rhadamanthus) 可靠地产生大量 (5-20 µL) 的毒液, 以响应刺激 (表 1)。一般而言, 大多数 peiratine、reduviine 和 harpactorine bug 会产生毒液来响应这种方法。在 stenopodaine bug 中, 刺激从Oncocephalus sp 中引发毒液, 而不是Thodelmus sp。取样的 holoptiline 和 emesine bug 并没有产生显著的毒液 (?...

Watch this Scientific Journal Video about Harvesting Venom Toxins from Assassin Bugs and Other Heteropteran Insects at JoVE.com

Harvesting Venom Toxins from Assassin Bugs and Other Heteropteran Insects

虽然亚目翅 (昆虫纲: 半翅目) 中的许多昆虫是有毒的, 但它们的毒液成分和毒液毒素的作用大多是未知的。本议定书描述了利用刺激、骚扰和腺体解剖来收获 heteropteran 毒液以进一步表征的方法。

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测定掠夺取食行为<em&gt; Pristionchus</em&gt;及其他线虫

This protocol provides multiple methods for the analysis and quantification of predatory feeding behaviors in nematodes. Many nematode species including ...

Extraction of Organochlorine Pesticides from Plastic Pellets and Plastic Type Analysis

从塑料颗粒中提取有机氯农药和塑料类型分析

Plastic resin pellets, categorized as microplastics (&#8804;5 mm in diameter), are small granules that can be unintentionally released to the environment ...

Extraction and Characterization of Surfactants from Atmospheric Aerosols

提取和表面活性剂的特性，从大气气溶胶

Surface-active compounds, or surfactants, present in atmospheric aerosols are expected to play important roles in the formation of liquid water clouds in ...

Methodology for Developing Life Tables for Sessile Insects in the Field Using the Whitefly, Bemisia tabaci, in Cotton As a Model System

Methodology for Developing Life Tables for Sessile Insects in the Field Using the Whitefly, Bemisia tabaci, in Cotton As a Model System

用虱、烟粉虱在棉花中开发无梗昆虫生命表的方法作为模型系统

Life tables provide a means of measuring the schedules of birth and death from populations over time. They also can be used to quantify the sources and ...

Composition and Properties of Aquafaba: Water Recovered from Commercially Canned Chickpeas

Aquafaba 的成分和性质: 从商用罐装鹰嘴中回收的水

Chickpea and other pulses are commonly sold as canned products packed in a thick solution or a brine. This solution has recently been shown to produce ...

Double-stranded RNA Oral Delivery Methods to Induce RNA Interference in Phloem and Plant-sap-feeding Hemipteran Insects

双链 rna 口服法诱导韧皮部和植物-sap 喂养 Hemipteran 昆虫 rna 干扰

Phloem and plant sap feeding insects invade the integrity of crops and fruits to retrieve nutrients, in the process damaging food crops. Hemipteran ...

Chemical Isolation, Quantification, and Separation of Skin Lipids from Reptiles

爬行动物皮肤脂类的化学分离、定量和分离

Reptiles signal to conspecifics using lipids in their skin, primarily to enable mate tracking and assessment. The isolation of these lipids has utility in ...

Cortisol Extraction from Sturgeon Fin and Jawbone Matrices

皮质醇从鱼翅和爪骨基质提取

The aims of this study were to develop a technique for the extraction of cortisol from sturgeon fins using two washing solvents (water and isopropanol) ...