| Establishing an Adult Mouse Brain Hippocampal Organotypic Slice Culture System that Allows for Tracing and Pharmacological Manipulation of ex vivo Neurogenesis
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Author:
Date:
2021-01-05
[Abstract] The function of the hippocampus depends on the process of adult hippocampal neurogenesis which underpins the exceptional neural plasticity of this structure, and is also frequently affected in CNS pathologies. Thus, manipulation of this process represents an important therapeutic goal. To identify potential strategies, organotypic adult brain slices are emerging as a valuable tool. Over the recent years, this methodology has been refined and here we present a combined protocol that brings together these refinements to enable long-term culture of adult hippocampal slices. We employ a sectioning technique that retains essential afferent inputs onto the hippocampus as well as serum-free culture conditions, so allowing an extended culture period. To sustain the neurogenic potential in the ...
[摘要] [摘要]海马的功能取决于成年海马神经发生的过程，该过程是这种结构异常的神经发育的基础，并且在中枢神经系统病理中也经常受到影响。因此，对该过程的操纵代表了重要的治疗目标。为了确定潜在的策略，器官型成人大脑切片正在成为一种有价值的工具。近年来，此方法已得到完善，在此我们提出一种组合协议， 汇集了这些改进，以实现成人海马切片的长期培养。我们采用了一种切片技术，可将必要的传入输入保留在海马上以及无血清培养条件下，因此可以延长培养时间。为了维持切片中的神经源性潜力，我们利用神经胶质生成抑制剂吲哚美辛。使用EdU保留分析使我们能够评估药理干预对神经发生的影响。通过这些改进，我们建立了一种简单可靠的方法来研究小分子/药物对离体增殖和神经元形成的影响，这将有助于未来发现驱动的药物筛选。
[背景技术]海马是具有高度的可塑性作为整个生命齿状回中正在进行的神经发生的结果，脑的独特区域。成年海马神经发生的这一过程始于在亚颗粒区（SGZ）中神经干细胞（NSC）的不对称分裂，该过程保留了干细胞池并生成了准备用于神经元分化的祖细胞（Kempermann等人，2004；Anacker和Hen ，2017； ...
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| A Protocol for Flavonols, Kaempferol and Quercetin, Staining in Plant Root Tips
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Author:
Date:
2020-10-05
[Abstract] Flavonols are a subclass of flavonoids of the group of plant secondary metabolites. In planta, flavonols play various functions such as antioxidant and natural regulator of auxin polar transport. Many lines of evidence have shown that flavonols also contribute to human health in anti-oxidation, anti-inflammation, and even prevention some types of cancer. Several methods have been utilized to measure flavonols such as high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), and diphenylboric acid-2-aminoethyl ester (DPBA) staining. While HPLC or LC-MS can quantitatively determine the level of flavonols, DPBA staining can provide an in-situ view of flavonols accumulation in the plants. In this protocol, a detailed procedure for ...
[摘要] [摘要]黄酮醇是植物次生代谢物中黄酮类的一个亚类。在植物中，黄酮醇具有多种功能，例如抗氧化剂和生长素极性转运的天然调节剂。许多证据表明，黄酮醇还有助于抗氧化，抗炎症甚至预防某些类型的癌症，从而对人类健康做出贡献。几种方法已用于测量黄酮醇，例如高效液相色谱（HPLC），液相色谱-质谱（LC-MS）和二苯硼酸-2-氨基乙基酯（DPBA）染色。HPLC或LC-MS可以定量测定黄酮醇的含量 DPBA染色可以提供植物中黄酮醇积累的原位视图。在该协议中，描述了对拟南芥根尖中的黄酮醇进行染色的详细步骤。将五天大的拟南芥幼苗浸入含有DPBA的溶液中，然后在共聚焦显微镜下观察黄酮醇（山ka酚和槲皮素）。
[背景]在拟南芥，黄酮醇是从缩合反应对香豆酰基辅酶A的一个分子和丙二酰-CoA的三个分子之间生物合成。Lewis等。（2011年）发现槲皮素而不是山奈酚是根型根茎植物生长素转运的抑制剂。此外，黄酮醇也可作为植物和人类的有效抗氧化剂。先前的研究调查了DPBA与黄酮醇相互作用时可以发出荧光（Sheahan和Rechnitz，1992）。基于此，DPBA已被广泛应用于检测植物中黄酮醇的积累（Nguyen等，2013、2015和2016 ； ...
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