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Single Cell Proteomics Using Frog (Xenopus laevis) Blastomeres Isolated from Early Stage Embryos, Which Form a Geometric Progression in Protein Content.
Sun L
,
Dubiak KM
,
Peuchen EH
,
Zhang Z
,
Zhu G
,
Huber PW
,
Dovichi NJ
.
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Single cell analysis is required to understand cellular heterogeneity in biological systems. We propose that single cells (blastomeres) isolated from early stage invertebrate, amphibian, or fish embryos are ideal model systems for the development of technologies for single cell analysis. For these embryos, although cell cleavage is not exactly symmetric, the content per blastomere decreases roughly by half with each cell division, creating a geometric progression in cellular content. This progression forms a ladder of single-cell targets for the development of successively higher sensitivity instruments. In this manuscript, we performed bottom-up proteomics on single blastomeres isolated by microdissection from 2-, 4-, 8-, 16-, 32-, and 50-cell Xenopus laevis (African clawed frog) embryos. Over 1 400 protein groups were identified in single-run reversed-phase liquid chromatography-electrospray ionization-tandem mass spectrometry from single balstomeres isolated from a 16-cell embryo. When the mass of yolk-free proteins in single blastomeres decreased from ∼0.8 μg (16-cell embryo) to ∼0.2 μg (50-cell embryo), the number of protein group identifications declined from 1 466 to 644. Around 800 protein groups were quantified across four blastomeres isolated from a 16-cell embryo. By comparing the protein expression among different blastomeres, we observed that the blastomere-to-blastomere heterogeneity in 8-, 16-, 32-, and 50-cell embryos increases with development stage, presumably due to cellular differentiation. These results suggest that comprehensive quantitative proteomics on single blastomeres isolated from these early stage embryos can provide valuable insights into cellular differentiation and organ development.
Aerts,
Patch clamp electrophysiology and capillary electrophoresis-mass spectrometry metabolomics for single cell characterization.
2014, Pubmed
Aerts,
Patch clamp electrophysiology and capillary electrophoresis-mass spectrometry metabolomics for single cell characterization.
2014,
Pubmed
Bendall,
From single cells to deep phenotypes in cancer.
2012,
Pubmed
Chen,
Ultrasensitive Proteome Profiling for 100 Living Cells by Direct Cell Injection, Online Digestion and Nano-LC-MS/MS Analysis.
2015,
Pubmed
Chen,
Single-Cell Analysis Using Drop-on-Demand Inkjet Printing and Probe Electrospray Ionization Mass Spectrometry.
2016,
Pubmed
Chen,
Simple and Integrated Spintip-Based Technology Applied for Deep Proteome Profiling.
2016,
Pubmed
Cohen,
Chemical cytometry: fluorescence-based single-cell analysis.
2008,
Pubmed
Cox,
Accurate proteome-wide label-free quantification by delayed normalization and maximal peptide ratio extraction, termed MaxLFQ.
2014,
Pubmed
Cox,
MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification.
2008,
Pubmed
Giesen,
Highly multiplexed imaging of tumor tissues with subcellular resolution by mass cytometry.
2014,
Pubmed
Kulak,
Minimal, encapsulated proteomic-sample processing applied to copy-number estimation in eukaryotic cells.
2014,
Pubmed
Li,
An Integrated Platform for Isolation, Processing, and Mass Spectrometry-based Proteomic Profiling of Rare Cells in Whole Blood.
2015,
Pubmed
Lombard-Banek,
Single-Cell Mass Spectrometry for Discovery Proteomics: Quantifying Translational Cell Heterogeneity in the 16-Cell Frog (Xenopus) Embryo.
2016,
Pubmed
,
Xenbase
Lorenzo Tejedor,
In situ molecular analysis of plant tissues by live single-cell mass spectrometry.
2012,
Pubmed
Lowry,
Enzyme patterns in single human muscle fibers.
1978,
Pubmed
Ludwig,
Over 2300 phosphorylated peptide identifications with single-shot capillary zone electrophoresis-tandem mass spectrometry in a 100 min separation.
2015,
Pubmed
Mellors,
Integrated microfluidic device for automated single cell analysis using electrophoretic separation and electrospray ionization mass spectrometry.
2010,
Pubmed
Moody,
Fates of the blastomeres of the 16-cell stage Xenopus embryo.
1987,
Pubmed
,
Xenbase
Murgia,
Single muscle fiber proteomics reveals unexpected mitochondrial specialization.
2015,
Pubmed
Nemes,
Metabolic differentiation of neuronal phenotypes by single-cell capillary electrophoresis-electrospray ionization-mass spectrometry.
2011,
Pubmed
Ong,
Classification of Large Cellular Populations and Discovery of Rare Cells Using Single Cell Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry.
2015,
Pubmed
Onjiko,
Single-cell mass spectrometry reveals small molecules that affect cell fates in the 16-cell embryo.
2015,
Pubmed
,
Xenbase
Peuchen,
Optimization and comparison of bottom-up proteomic sample preparation for early-stage Xenopus laevis embryos.
2016,
Pubmed
,
Xenbase
Rubakhin,
Profiling metabolites and peptides in single cells.
2011,
Pubmed
,
Xenbase
Rubakhin,
Quantitative measurements of cell-cell signaling peptides with single-cell MALDI MS.
2008,
Pubmed
Schubert,
Single-cell analysis: The deepest differences.
2011,
Pubmed
Smith,
An accurate mass tag strategy for quantitative and high-throughput proteome measurements.
2002,
Pubmed
Smits,
Global absolute quantification reveals tight regulation of protein expression in single Xenopus eggs.
2014,
Pubmed
,
Xenbase
Sun,
Coupling methanol denaturation, immobilized trypsin digestion, and accurate mass and time tagging for liquid-chromatography-based shotgun proteomics of low nanogram amounts of RAW 264.7 cell lysate.
2012,
Pubmed
Sun,
Quantitative proteomics of Xenopus laevis embryos: expression kinetics of nearly 4000 proteins during early development.
2014,
Pubmed
,
Xenbase
Sun,
Ultrasensitive and fast bottom-up analysis of femtogram amounts of complex proteome digests.
2013,
Pubmed
Sun,
Integrated capillary zone electrophoresis-electrospray ionization tandem mass spectrometry system with an immobilized trypsin microreactor for online digestion and analysis of picogram amounts of RAW 264.7 cell lysate.
2013,
Pubmed
Tian,
Rare cell proteomic reactor applied to stable isotope labeling by amino acids in cell culture (SILAC)-based quantitative proteomics study of human embryonic stem cell differentiation.
2011,
Pubmed
Waanders,
Quantitative proteomic analysis of single pancreatic islets.
2009,
Pubmed
Wang,
Development of mass spectrometry-based shotgun method for proteome analysis of 500 to 5000 cancer cells.
2010,
Pubmed
Wang,
Improving the comprehensiveness and sensitivity of sheathless capillary electrophoresis-tandem mass spectrometry for proteomic analysis.
2012,
Pubmed
Wei,
Pulsed Direct Current Electrospray: Enabling Systematic Analysis of Small Volume Sample by Boosting Sample Economy.
2015,
Pubmed
Whittal,
Nanoliter chemistry combined with mass spectrometry for peptide mapping of proteins from single mammalian cell lysates.
1998,
Pubmed
Wiśniewski,
High recovery FASP applied to the proteomic analysis of microdissected formalin fixed paraffin embedded cancer tissues retrieves known colon cancer markers.
2011,
Pubmed
Wiśniewski,
Universal sample preparation method for proteome analysis.
2009,
Pubmed
Zhang,
Nearly 1000 Protein Identifications from 50 ng of Xenopus laevis Zygote Homogenate Using Online Sample Preparation on a Strong Cation Exchange Monolith Based Microreactor Coupled with Capillary Zone Electrophoresis.
2016,
Pubmed
,
Xenbase
