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通过可视化基因和蛋白质活性以及定量维生素A代谢物来识别维生素A信号传导。

Identifying vitamin A signaling by visualizing gene and protein activity, and by quantification of vitamin A metabolites.

作者信息

Shannon Stephen R, Yu Jianshi, Defnet Amy E, Bongfeldt Danika, Moise Alexander R, Kane Maureen A, Trainor Paul A

机构信息

Stowers Institute for Medical Research, Kansas City, MO, United States; University of Kansas Medical Center, Department of Anatomy and Cell Biology, Kansas City, KS, United States.

University of Maryland Baltimore, School of Pharmacy, Department of Pharmaceutical Sciences, Baltimore, MD, United States.

出版信息

Methods Enzymol. 2020;637:367-418. doi: 10.1016/bs.mie.2020.03.011. Epub 2020 Apr 21.

DOI:10.1016/bs.mie.2020.03.011
PMID:32359653
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7565286/
Abstract

Vitamin A (retinol) is an essential nutrient for embryonic development and adult homeostasis. Signaling by vitamin A is carried out by its active metabolite, retinoic acid (RA), following a two-step conversion. RA is a small, lipophilic molecule that can diffuse from its site of synthesis to neighboring RA-responsive cells where it binds retinoic acid receptors within RA response elements of target genes. It is critical that both vitamin A and RA are maintained within a tight physiological range to protect against developmental disorders and disease. Therefore, a series of compensatory mechanisms exist to ensure appropriate levels of each. This strict regulation is provided by a number synthesizing and metabolizing enzymes that facilitate the precise spatiotemporal control of vitamin A metabolism, and RA synthesis and signaling. In this chapter we describe protocols that (1) biochemically isolate and quantify vitamin A and its metabolites and (2) visualize the spatiotemporal activity of genes and proteins involved in the signaling pathway.

摘要

维生素A（视黄醇）是胚胎发育和成人内环境稳态所必需的营养素。维生素A通过其活性代谢物视黄酸（RA）进行两步转化后发挥信号传导作用。RA是一种小分子亲脂性分子，它可以从合成部位扩散到邻近的RA反应性细胞，在那里它与靶基因RA反应元件内的视黄酸受体结合。维生素A和RA都必须维持在严格的生理范围内，以预防发育障碍和疾病，这一点至关重要。因此，存在一系列补偿机制来确保每种物质的适当水平。这种严格的调控由多种合成和代谢酶提供，这些酶有助于对维生素A代谢、RA合成和信号传导进行精确的时空控制。在本章中，我们描述了一些实验方案，这些方案（1）从生物化学角度分离和定量维生素A及其代谢物，（2）可视化信号通路中涉及的基因和蛋白质的时空活性。

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SABER amplifies FISH: enhanced multiplexed imaging of RNA and DNA in cells and tissues.SABER 增强 FISH：在细胞和组织中增强 RNA 和 DNA 的多重成像。
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