A Simple and Effective Sample Preparation Strategy for MALDI-MS Imaging of Neuropeptide Changes in the Crustacean Brain Due to Hypoxia and Hypercapnia Stress
A Simple and Effective Sample Preparation Strategy for MALDI-MS Imaging of Neuropeptide Changes in the Crustacean Brain Due to Hypoxia and Hypercapnia Stress
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DOI：
10.1021/jasms.9b00107
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发表时间：
2020-05-06
影响因子：
3.2
通讯作者：
Li, Lingjun
中科院分区：
文献类型：
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作者：
Buchberger, Amanda R.;Vu, Nhu Q.;Li, Lingjun
基质辅助激光解吸/电离（MALDI）-MS成像已被用于成像各种生物分子，包括神经肽。清洗组织切片是消除干扰背景和改善低浓度目标分析物分子检测的有效方法;然而，许多先前的方法尚未通过MALDI-MS成像测试用于神经肽分析。使用甲壳类动物作为神经模式生物，我们开发了一种新的，简单的洗涤程序，并应用这种方法来表征由于缺氧应激神经肽的变化。用10 s 50：50 EtOH：H2O洗涤，神经肽覆盖率提高了1.15倍，而归一化信号强度增加了5.28倍。具体而言，缺氧和高碳酸血症的压力条件进行了调查，由于其环境相关的海洋无脊椎动物。许多神经肽，包括RFamides，pyrokinin，和心脏活性肽，表现出不同的上调和下调特定的神经肽亚型。由于甲壳类动物的神经肽是同源的那些在人类中发现的，这些研究的结果可以应用于了解参与医疗缺氧和高碳酸血症的神经肽的潜在作用。
Matrix-assisted laser desorption/ionization (MALDI)-MS imaging has been utilized to image a variety of biomolecules, including neuropeptides. Washing a tissue section is an effective way to eliminate interfering background and improve detection of low concentration target analyte molecules; however, many previous methods have not been tested for neuropeptide analysis via MALDI-MS imaging. Using crustaceans as a neurological model organism, we developed a new, simple washing procedure and applied this method to characterize neuropeptide changes due to hypoxia stress. With a 10 s 50:50 EtOH:H2O wash, neuropeptide coverage was improved by 1.15-fold, while normalized signal intensities were increased by 5.28-fold. Specifically, hypoxia and hypercapnia stress conditions were investigated due to their environmental relevance to marine invertebrates. Many neuropeptides, including RFamides, pyrokinin, and cardioactive peptides, showed distinct up- and down-regulation for specific neuropeptide isoforms. Since crustacean neuropeptides are homologous to those found in humans, results from these studies can be applied to understand potential roles of neuropeptides involved in medical hypoxia and hypercapnia.