TY - JOUR
T1 - Direct profiling of the phospholipid composition of adult Caenorhabditis elegans using whole-body imaging mass spectrometry
AU - Hameed, Saira
AU - Ikegami, Koji
AU - Sugiyama, Eiji
AU - Matsushita, Shoko
AU - Kimura, Yoshishige
AU - Hayasaka, Takahiro
AU - Sugiura, Yuki
AU - Masaki, Noritaka
AU - Waki, Michihiko
AU - Ohta, Isao
AU - Hossen, Md Amir
AU - Setou, Mitsutoshi
N1 - Funding Information:
Acknowledgments We would like to thank Drs. Naoko Goto-Inoue and Dan Nicolaescu for their critical reading of the manuscript and constructive comments. The nematode strains used in this work were donated by the Caenorhabditis Genetics Center, which is funded by the National Center for Research Resources, National Institutes of Health (NIH). This work was supported by JSPS KAKENHI for Scientific Research on Innovative Areas “Machinery of bioactive lipids in homeostasis and disease” (25116712) and for Kiban-B (25293044) (to M.S.), by JSPS KAKENHI for Kiban-C (25440082) (to Y.K.) and by the MEXT Project for Creation of Research Platforms and Sharing of Advanced Research Infrastructure. S.H. is receiving a scholarship from MEXT and formerly from JASSO and the Otsuka Toshimi Foundation.
Publisher Copyright:
© The Author(s) 2015.
PY - 2017/10
Y1 - 2017/10
N2 - 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 freezecracked 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.
AB - 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 freezecracked 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.
KW - Caenorhabditis elegans
KW - Cuticle
KW - Exoskeleton
KW - Freeze-cracking
KW - Matrix-assisted laser desorption/ ionization-imaging mass spectrometry
KW - Phosphatidylcholine
KW - Phosphatidylinositol
UR - http://www.scopus.com/inward/record.url?scp=84940093295&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84940093295&partnerID=8YFLogxK
U2 - 10.1007/s00216-015-8932-7
DO - 10.1007/s00216-015-8932-7
M3 - Article
C2 - 26310845
AN - SCOPUS:84940093295
SN - 1618-2642
VL - 407
SP - 7589
EP - 7602
JO - Analytical and bioanalytical chemistry
JF - Analytical and bioanalytical chemistry
IS - 25
ER -