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Student Fine Scissors

Fine剪刀

Company: Fine Science Tools
Catalog#: 91460-11
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Generation of Mouse Primary Hypothalamic Neuronal Cultures for Circadian Bioluminescence Assays
Author:
Date:
2021-03-05
[Abstract]  

An endogenous circadian clock system enables organisms to adapt to time-of-day dependent environmental changes. In consequence, most physiological processes exhibit daily rhythms of, e.g., energy metabolism, immune function, sleep, or hormone production. Hypothalamic circadian clocks have been identified to play a particular role in coordinating many of these processes. Primary neuronal cultures are widely used as a physiologically relevant model to study molecular events within neurons. However, as circadian rhythms include dynamic molecular changes over longer timescales that vary between individual cells, longitudinal measurement methods are essential to investigate the regulation of circadian clocks of hypothalamic neurons. Here we provide a protocol for generating primary

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[摘要]  [摘要]内源性生物钟系统使生物能够适应与时间相关的环境变化。结果,大多数生理过程表现出例如能量代谢,免疫功能,睡眠或激素产生的每日节律。下丘脑生物钟已被确认在协调许多这些过程中起特定作用。 原代神经元文化被广泛用作研究神经元内分子事件的生理相关模型。然而,由于昼夜节律包括较长时间范围内的动态分子变化,而这种变化在各个细胞之间会有所不同,因此纵向测量方法对于研究下丘脑神经元昼夜节律的调节至关重要。在这里,我们提供了用于生成表达昼夜节律性荧光素酶报道基因的下丘脑神经元文化的协议。通过执行生物发光测量,此类报告细胞可用于以高时间分辨率纵向监测细胞昼夜节律。


[背景]为了适应重复在其环境中的时间-日期依赖性变化,许多生物已开发出一种内源性生物钟系统调节行为和生理过程的24小时的节律(夏尔马,2003)。在哺乳动物中,一个昼夜节律性起搏器主要位于下丘脑上视交叉上核(SCN)。它与外部时间协调整个身体的细胞时钟调节。睡眠,食欲和新陈代谢的每日模式由下丘脑神经元中的细胞昼夜节律调节(Cedernaes等,2019)。

在哺乳动物细胞中,昼夜节律时钟由互锁的转录-翻译反馈环(TTFL)组成。在核心TTFL中,转录因子昼夜运动输出周期kaput(CLOCK)和脑和肌肉芳基碳氢化合物受体核转运蛋白样蛋白1(BMAL1或ARNTL)激活其自身阻遏物,周期(PER1-3)和隐色蛋白的表达(CRY1 ...

Confocal Microscopy of Reovirus Transport in Living Dorsal Root Ganglion Neurons
Author:
Date:
2020-11-20
[Abstract]  

Neurotropic reoviruses repurpose host machinery to traffic over long distances in neuronal processes and access distal replication sites. Understanding mechanisms of neuronal transmission is facilitated by using simplified in vitro primary neuronal culture models. Advances in the design of compartmentalized microfluidic devices lend robustness to neuronal culture models by enabling compartmentalization and manipulation of distinct neuronal processes. Here, we describe a streamlined methodology to culture sensory neurons dissociated from dorsal root ganglia of embryonic rats in microfluidic devices. We further describe protocols to exogenously label reovirus and image, track, and analyze transport of single reovirus particles in living neurons. These techniques can be adapted to study

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[摘要]  [摘要] 嗜神经性呼肠孤病毒重新利用宿主机器在神经元过程中进行长距离的传输,并进入远端复制部位。简化的体外原代神经元培养模型有助于理解神经元传递机制。室化微流控装置设计的进展使得不同的神经元过程能够被划分和操作,从而为神经元培养模型提供了稳健性。在这里,我们描述了一种在微流控装置中培养胚胎大鼠背根神经节分离的感觉神经元的方法。我们进一步描述了外源性标记呼肠孤病毒的方法,并对单个呼肠孤病毒粒子在活神经元中的转运进行了成像、跟踪和分析。这些技术可应用于研究其他嗜神经病毒的轴突定向转运以及参与信号传导和病理学的神经因子。



[背景]来自不同家族的病毒,包括黄病毒科、疱疹病毒科、小角RNA病毒科和弹状病毒科,突破神经系统的保护屏障,造成严重的疾病和经济负担(Koyuncu等人,2013年;Bohmwald等人,2018年;Tyler,2018年)。哺乳动物正呼肠孤病毒(reovirus,reovirus)属于呼肠孤病毒科,在多种年轻哺乳动物中引起血清型依赖性神经元感染,可导致致命性脑炎(Tyler等人,1986年;Dermody等人,2013年)。呼肠孤病毒没有被两个同心蛋白壳包裹的片段dsRNA基因组包裹,是研究神经系统病毒感染的一种灵活工具(Dermody等人,2013年)。虽然呼肠孤病毒感染的细胞和分子机制已被广泛地利用转化细胞系进行研究,但这些系统并不能捕捉到极化神经元细胞的复杂性。感染神经元的病毒必须在轴突中长距离传播,才能到达复制和释放的远端。为了了解呼肠孤病毒进入神经元和长距离运输的机制,我们最近采用了一些技术来培养原代神经元,并对活细胞中荧光标记的呼肠孤病毒成像(Aravamudhan等人,2020年)。 ...

Transplantation of Fecal Microbiota Shaped by Diet
Author:
Date:
2018-01-05
[Abstract]  Alterations in diet and gut microbial ecology underlie the pathogenesis of type 1 diabetes (T1D). In the non-obese diabetic (NOD) mouse, we found high concentrations of bacterial metabolites acetate and butyrate in blood and faeces correlated with protection from disease. We reconstituted germ free (GF) NOD mice with fecal bacteria from protected NOD mice fed with high acetate- and butyrate-yielding diets, to test whether the transferred gut microbiota protect against the development of T1D. GF NOD mice that received a microbiota shaped by high acetate- but not butyrate-yielding diet showed a marked protection against diabetes. This fecal transplantation assay demonstrated the potential for a dietary technology to reshape the gut microbiota that enables specific bacteria to transfer ... [摘要]  饮食和肠道微生物生态学的改变是1型糖尿病(T1D)发病的基础。 在非肥胖糖尿病(NOD)小鼠中,我们发现血液和粪便中高浓度的乙酸和丁酸的细菌代谢物与防止疾病相关。 我们使用来自受保护的NOD小鼠的粪便细菌重建无菌(GF)NOD小鼠,饲喂高乙酸盐和丁酸盐产生的饮食,以测试转移的肠道微生物群是否防止T1D的发展。 GF NOD小鼠接受由高乙酸盐但不是丁酸盐产生饮食形成的微生物群显示出对糖尿病的显着保护。 这种粪便移植试验表明,饮食技术可以重塑肠道微生物群,使特定的细菌能够转移对T1D的保护。


【背景】已经在各种各样的疾病和病况中观察到肠道微生物群的变化。由于共生菌和致病菌之间的比例变化,肠道益生菌微生物群体失去了体内平衡。当微生物的组成变化很大时,可以观察到生物异常,某些种类的数量增加或减少(Clemente等,2012; Rajilic-Stojanovic,2013)。此外,肠道微生物群组成(其可能影响代谢物如SCFAs)的变化与许多炎性疾病有关(Clemente等人,2012),包括T1D(de Goffau等人 >,2013; Endesfelder ...

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