High-Sensitivity Mass Spectrometry for Probing Gene Translation in Single Embryonic Cells in the Early Frog (Xenopus) Embryo.

Camille Lombard-Banek,Sally A Moody,Peter Nemes

doi:10.3389/fcell.2016.00100

Camille Lombard-Banek, Sally A Moody + Show 1 more

Open Access

https://doi.org/10.3389/fcell.2016.00100

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Abstract

Direct measurement of protein expression with single-cell resolution promises to deepen the understanding of the basic molecular processes during normal and impaired development. High-resolution mass spectrometry provides detailed coverage of the proteomic composition of large numbers of cells. Here we discuss recent mass spectrometry developments based on single-cell capillary electrophoresis that extend discovery proteomics to sufficient sensitivity to enable the measurement of proteins in single cells. The single-cell mass spectrometry system is used to detect a large number of proteins in single embryonic cells in the 16-cell embryo of the South African clawed frog (Xenopus laevis) that give rise to distinct tissue types. Single-cell measurements of protein expression provide complementary information on gene transcription during early development of the vertebrate embryo, raising a potential to understand how differential gene expression coordinates normal cell heterogeneity during development.

Highlights

Single-cell analysis technologies are essential to understanding cell heterogeneity during normal development and disease
We have recently demonstrated Label-free quantification (LFQ) for single blastomeres of neural fates in the 16-cell embryo using the protocol presented here (Lombard-Banek et al, 2016b)
Figures adapted with permission from Lombard-Banek et al (2016a,b). These proteins are involved in brain development (Pera et al, 2002; Seo et al, 2005), which is the stereotypical fate of D11 cells (Moody, 1987a)

Summary

INTRODUCTION

Single-cell analysis technologies are essential to understanding cell heterogeneity during normal development and disease. Quantification of ∼150 different proteins between the blastomeres has captured translational cell heterogeneity in the 16-cell vertebrate embryo (Lombard-Banek et al, 2016a) These results complement known transcriptional cell differences in the embryo, and provide previously unknown details on how differential gene expression establishes cell heterogeneity during early embryonic development. In this contribution, we give an overview of the major steps of the single-cell CE-MS workflow (Figure 1).

PROCEDURES

Anticipated Results

CONCLUSIONS