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使用全身成像质谱法直接分析成年秀丽隐杆线虫的磷脂组成。

Direct profiling of the phospholipid composition of adult Caenorhabditis elegans using whole-body imaging mass spectrometry.

作者信息

Hameed Saira, Ikegami Koji, Sugiyama Eiji, Matsushita Shoko, Kimura Yoshishige, Hayasaka Takahiro, Sugiura Yuki, Masaki Noritaka, Waki Michihiko, Ohta Isao, Hossen Md Amir, Setou Mitsutoshi

机构信息

Department of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu, Shizuoka, 431-3192, Japan.

Faculty of Health Sciences, Health Innovation and Technology Center, Hokkaido University, Kita 12 Nishi 5, Kita-ku, Sapporo, Hokkaido, 060-0812, Japan.

出版信息

Anal Bioanal Chem. 2015 Oct;407(25):7589-602. doi: 10.1007/s00216-015-8932-7. Epub 2015 Aug 27.

DOI:10.1007/s00216-015-8932-7
PMID:26310845
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4575384/
Abstract

A protocol for the direct analysis of the phospholipid composition in the whole body of adult soil nematode, Caenorhabditis elegans (C. elegans), was developed, which combined freeze-cracking of the exoskeletal cuticle and matrix-assisted laser desorption/ionization-imaging mass spectrometry (MALDI-IMS). Biomolecules in the m/z range from 700 to 900 were more effectively detected in the freeze-cracked than from simple frozen adult nematode bodies. Different distribution of biomolecules was observed in a nematode body when the matrix was applied with a sublimation deposition method. The whole-body IMS technique was applied on genetically deficient mutant C. elegans to combine whole-body lipidomics and genetics, by comparing the fatty acid compositions, especially of the phosphatidylcholine (PC) species, between the wild-type and fat-1 mutants, which lack the gene encoding an n-3 fatty acid desaturase. A significant reduction of PC(20:5/20:5) and PC(20:4/20:5) and a marked increase of PC(20:4/20:4), PC(20:3/20:4), and PC(20:3/20:3) were detected in the fat-1 mutants in positive ion mode. In addition, phospholipid compositions other than PCs were analyzed in negative ion mode. A loss of a possible phosphatidylinositol (PI) with 18:0/20:5 and a compensative accumulation of putative PI(18:0/20:4) were detected in the fat-1 mutants. In conclusion, the whole-body MALDI-IMS technique is useful for the profiling of multiple biomolecules in C. elegans in both intra- and inter-individual levels.

摘要

我们开发了一种直接分析成年土壤线虫秀丽隐杆线虫（C. elegans）全身磷脂组成的方法，该方法结合了外骨骼角质层的冷冻裂解和基质辅助激光解吸/电离成像质谱（MALDI-IMS）。与简单冷冻的成年线虫相比，在冷冻裂解的线虫中，更有效地检测到了质荷比在700至900范围内的生物分子。当采用升华沉积法施加基质时，在线虫体内观察到了生物分子的不同分布。通过比较野生型和缺乏编码n-3脂肪酸去饱和酶基因的fat-1突变体之间的脂肪酸组成，特别是磷脂酰胆碱（PC）种类，将全身IMS技术应用于基因缺陷的秀丽隐杆线虫突变体，以结合全身脂质组学和遗传学。在正离子模式下，在fat-1突变体中检测到PC(20:5/20:5)和PC(20:4/20:5)显著减少，PC(20:4/20:4)、PC(20:3/20:4)和PC(20:3/20:3)显著增加。此外，在负离子模式下分析了除PCs之外的磷脂组成。在fat-1突变体中检测到一种可能的18:0/20:5磷脂酰肌醇（PI）的缺失和假定的PI(18:0/20:4)的补偿性积累。总之，全身MALDI-IMS技术可用于在个体内和个体间水平上对秀丽隐杆线虫中的多种生物分子进行分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/993c/4575384/99e4f4ea0917/216_2015_8932_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/993c/4575384/33b83b91ba37/216_2015_8932_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/993c/4575384/b09b268107b9/216_2015_8932_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/993c/4575384/7e54c9bdf35c/216_2015_8932_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/993c/4575384/189f7de82b23/216_2015_8932_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/993c/4575384/99e4f4ea0917/216_2015_8932_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/993c/4575384/33b83b91ba37/216_2015_8932_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/993c/4575384/b09b268107b9/216_2015_8932_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/993c/4575384/7e54c9bdf35c/216_2015_8932_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/993c/4575384/189f7de82b23/216_2015_8932_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/993c/4575384/99e4f4ea0917/216_2015_8932_Fig5_HTML.jpg

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