Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 60, Issue 23, Pages 12852-12858Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.202100923
Keywords
embryos; mass spectrometry; metabolites; proteins; single cells
Categories
Funding
- Arnold and Mabel Beckman Foundation Beckman Young Investigator award
- National Science Foundation [DBI-1826932, IOS-1832968]
- National Institutes of Health [1R35GM124755]
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The study reports the development of in vivo subcellular high-resolution mass spectrometry for proteo-metabolomic molecular systems biology in complex tissues. Through precision-translated fabricated microcapillaries, subcellular content of different cell types was analyzed with differential quantitative activities of proteins and metabolites identified. The technology preserved integrity of analyzed cells, neighboring cells, and embryos, with 95% of analyzed embryos developing into sentient tadpoles indistinguishable from wild-type siblings.
We report the development of in vivo subcellular high-resolution mass spectrometry (HRMS) for proteo-metabolomic molecular systems biology in complex tissues. With light microscopy, we identified the left-dorsal and left-ventral animal cells in cleavage-stage non-sentient Xenopus laevis embryos. Using precision-translated fabricated microcapillaries, the subcellular content of each cell was double-probed, each time swiftly (<5 s/event) aspirating <5 % of cell volume (approximate to 10 nL). The proteins and metabolites were analyzed by home-built ultrasensitive capillary electrophoresis electrospray ionization employing orbitrap or time-of-flight HRMS. Label-free detection of approximate to 150 metabolites (57 identified) and 738 proteins found proteo-metabolomic networks with differential quantitative activities between the cell types. With spatially and temporally scalable sampling, the technology preserved the integrity of the analyzed cells, the neighboring cells, and the embryo. 95 % of the analyzed embryos developed into sentient tadpoles that were indistinguishable from their wild-type siblings based on anatomy and visual function in a background color preference assay.
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