Cell lineage-guided mass spectrometry reveals increased energy metabolism and reactive oxygen species in the vertebrate organizer

Authors

Aparna B. Baxi, Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742.
Jie Li, Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742.
Vi M. Quach, Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742.
Leena R. Pade, Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742.
Sally A. Moody, Department of Anatomy and Cell Biology, School of Medical and Health Sciences, The George Washington University, Washington, DC 20037.
Peter Nemes, Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742.

Document Type

Journal Article

Publication Date

2-6-2024

Journal

Proceedings of the National Academy of Sciences of the United States of America

Volume

121

Issue

6

DOI

10.1073/pnas.2311625121

Keywords

Spemann–Mangold Organizer; Xenopus; mass spectrometry; metabolomics; proteomics

Abstract

Molecular understanding of the vertebrate Organizer, a tissue center critical for inductive signaling during gastrulation, has so far been mostly limited to transcripts and a few proteins, the latter due to limitations in detection and sensitivity. The Spemann-Mangold Organizer (SMO) in the South African Clawed Frog (), a popular model of development, has long been known to be the origin of signals that pattern the mesoderm and central nervous system. Molecular screens of the SMO have identified several genes responsible for the ability of the SMO to establish the body axis. Nonetheless, a comprehensive study of proteins and metabolites produced specifically in the SMO and their functional roles has been lacking. Here, we pioneer a deep discovery proteomic and targeted metabolomic screen of the SMO in comparison to the remainder of the embryo using high-resolution mass spectrometry (HRMS). Quantification of ~4,600 proteins and a panel of targeted metabolites documented differential expression for 460 proteins and multiple intermediates of energy metabolism in the SMO. Upregulation of oxidative phosphorylation and redox regulatory proteins gave rise to elevated oxidative stress and an accumulation of reactive oxygen species in the SMO. Imaging experiments corroborated these findings, discovering enrichment of hydrogen peroxide in the SMO. Chemical perturbation of the redox gradient perturbed mesoderm involution during early gastrulation. HRMS expands the bioanalytical toolbox of cell and developmental biology, providing previously unavailable information on molecular classes to challenge and refine our classical understanding of the Organizer and its function during early patterning of the embryo.

APA Citation

Baxi, Aparna B.; Li, Jie; Quach, Vi M.; Pade, Leena R.; Moody, Sally A.; and Nemes, Peter, "Cell lineage-guided mass spectrometry reveals increased energy metabolism and reactive oxygen species in the vertebrate organizer" (2024). GW Authored Works. Paper 4384.
https://hsrc.himmelfarb.gwu.edu/gwhpubs/4384

Department

Anatomy and Regenerative Biology