| A Blood-retina Barrier Permeability Assay in Young Mice Using Sulfo-NHS-LC-biotin Perfusion
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Author:
Date:
2018-10-20
[Abstract] Brain and retinal vasculatures exhibit restricted vascular permeability known as blood-brain barrier and blood-retina barrier. Vascular permeability can be evaluated by perfusion of the amine reactive ester derivatives of biotin such as sulfo-NHS-LC-biotin. This protocol describes experimental procedures of sulfo-NHS-LC-biotin perfusion to evaluate retinal vascular permeability. Perfused sulfo-NHS-LC-biotin remained within vessels in wild-type postnatal day 15 (P15) retinas, confirming an intact blood-retina barrier. In contrast, sulfo-NHS-LC-biotin was occasionally detected in extravascular spaces in perfused Eogt−/− retinas suggesting a partly impaired vascular integrity in the absence of Eogt (Sawaguchi et al., 2017).
[摘要] 脑和视网膜脉管系统表现出受限的血管通透性，称为血脑屏障和血 - 视网膜屏障。 血管通透性可以通过灌注生物素的胺反应性酯衍生物如磺基-NHS-LC-生物素来评估。 该方案描述了磺基-NHS-LC-生物素灌注的实验程序，以评估视网膜血管通透性。 灌注的磺基-NHS-LC-生物素保留在野生型出生后第15天（P15）视网膜的血管内，证实了完整的血 - 视网膜屏障。 相比之下，在灌注的 Eogt - / - >视网膜中，偶尔会在血管外空间检测到磺基-NHS-LC-生物素，这表明在没有的情况下血管完整性部分受损。 Eogt >（Sawaguchi et al。>，2017）。
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| Dissection and Whole Mount Staining of Retina from Neonatal Mice
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Author:
Date:
2018-10-05
[Abstract] Here we provide a detailed protocol for whole mount staining of mouse retina. This protocol was used to analyze retinal angiogenesis in newborn mice (Sawaguchi et al., 2017) by modifying the original protocols (Powner et al., 2012; Tual-Chalot et al., 2013). This protocol can also be used for whole mount staining of adult retina.
[摘要] 在这里，我们提供了小鼠视网膜整体染色的详细方案。 该方案用于分析新生小鼠的视网膜血管生成（Sawaguchi et al。，2017），修改原始方案（Powner et al。，2012; Tual-Chalot et al。，2013）。 该方案也可用于成人视网膜的整体染色。
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| Assaying Mechanonociceptive Behavior in Drosophila Larvae
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Author:
Date:
2018-02-20
[Abstract] Drosophila melanogaster larvae have been extensively used as a model to study the molecular and cellular basis of nociception. The larval nociceptors, class IV dendritic arborization (C4da) neurons, line the body wall of the animal and respond to various stimuli including noxious heat and touch. Activation of C4da neurons results in a stereotyped escape behavior, characterized by a 360° rolling response along the body axis followed by locomotion speedup. The genetic accessibility of Drosophila has allowed the identification of mechanosensory channels and circuit elements required for nociceptive responses, making it a useful and straightforward readout to understand the cellular and molecular basis of nociceptive function and behavior. We have optimized the protocol to ...
[摘要] 果蝇幼虫已广泛用作研究伤害感受的分子和细胞基础的模型。 幼虫伤害感受器，IV类树突状树枝状（C4da）神经元，排列在动物体壁上，并对各种刺激作出反应，包括有害的热和触觉。 激活C4da神经元导致刻板脱逃行为，其特征在于沿着身体轴线的360°滚动响应，随后是运动加速。 果蝇的遗传可及性允许识别伤害性反应所需的机械感受通道和电路元件，使其成为理解伤害性功能和行为的细胞和分子基础的有用和直接的读数。 我们已经优化了该协议以检测果蝇幼虫中的机械伤害行为。
【背景】伤害感受，是检测和避免有害刺激的先天能力，在整个动物界高度保守。果蝇幼虫能够检测并避免各种有害刺激，包括有害触觉，热和光（Tracey等人，2003; Hwang等人， ，2007; Xiang et al。，2010）。高于特定阈值（> 30mN）的机械刺激在所有幼虫阶段（Almeida-Carvalho等人，2017）都引发了定型滚动逃避反应，其被认为已经进化以避免通过寄生蜂如L。 boulardi （Hwang et。，2007）。这种逃避反应是由伤害性C4da神经元的激活介导的，伤害性C4da神经元具有覆盖整个体壁的感觉树突，允许动物检测有害线索。 C4da神经元表达属于DEG / ENaC家族的几种机械感受通道（Pickpocket [ppk]，ppk26 / ...
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