. This research team recently published the first quantitative proteomic analysis of a devel- oping embryo (Sci Rep. 2014; 4: 4365. PMID: 24626130). We studied single Xenopus laevis embryos at six stages of development. We initially quantified the expression change of 4,000 proteins across at least four stages of development; reanalysis of the data with an improved genomic database has extended our identifications by 50%. Data have been made available to the community on the Scien- tific Reports web site, at Xenbase, and at PeptideAtlas. We will extend our initial study by quantitatively monitoring the expression and phosphorylation status of proteins isolated from single Xenopus oocytes, eggs, and blastomeres. Quantitative protein expression changes will also be measured from cryosections perpendicular to developmental axes to identify patterning gradients, along a selected cell lineage, from an explant of that lineage, and in oocytes following nuclear transplantation. The results of these studies will be an unprecedented resource to the Xenopus community by providing detailed data on the evolution of protein expression during development, and chromatin reorganization in embryos and following nuclear transplantation. We will work with Xenbase, PeptideAtlas, and VectorBase to disseminate our results as widely as possible.

Public Health Relevance

. We will quantitatively analyze protein expression changes associated with pattern forma- tion and commitment to cell lineage in early stage Xenopus embryos. These studies are vital in our understanding of normal embryonic development and may provide insight into the direction of stem cells along specific lineages.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD084399-02
Application #
9116239
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Coulombe, James N
Project Start
2015-08-01
Project End
2020-04-30
Budget Start
2016-05-01
Budget End
2017-04-30
Support Year
2
Fiscal Year
2016
Total Cost
Indirect Cost
Name
University of Notre Dame
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
824910376
City
Notre Dame
State
IN
Country
United States
Zip Code
46556
Hayes, Michael H; Peuchen, Elizabeth H; Dovichi, Norman J et al. (2018) Dual roles for ATP in the regulation of phase separated protein aggregates in Xenopus oocyte nucleoli. Elife 7:
Sindelka, Radek; Abaffy, Pavel; Qu, Yanyan et al. (2018) Asymmetric distribution of biomolecules of maternal origin in the Xenopus laevis egg and their impact on the developmental plan. Sci Rep 8:8315
Lee-Liu, Dasfne; Sun, Liangliang; Dovichi, Norman J et al. (2018) Quantitative Proteomics After Spinal Cord Injury (SCI) in a Regenerative and a Nonregenerative Stage in the Frog Xenopus laevis. Mol Cell Proteomics 17:592-606
Peuchen, Elizabeth H; Cox, Olivia F; Sun, Liangliang et al. (2017) Phosphorylation Dynamics Dominate the Regulated Proteome during Early Xenopus Development. Sci Rep 7:15647
Peuchen, Elizabeth H; Zhu, Guije; Sun, Liangliang et al. (2017) Evaluation of a commercial electro-kinetically pumped sheath-flow nanospray interface coupled to an automated capillary zone electrophoresis system. Anal Bioanal Chem 409:1789-1795
Sun, Liangliang; Dubiak, Kyle M; Peuchen, Elizabeth H et al. (2016) Single Cell Proteomics Using Frog (Xenopus laevis) Blastomeres Isolated from Early Stage Embryos, Which Form a Geometric Progression in Protein Content. Anal Chem 88:6653-7
Peuchen, Elizabeth H; Sun, Liangliang; Dovichi, Norman J (2016) Optimization and comparison of bottom-up proteomic sample preparation for early-stage Xenopus laevis embryos. Anal Bioanal Chem 408:4743-9
Sun, Liangliang; Champion, Matthew M; Huber, Paul W et al. (2016) Proteomics of Xenopus development. Mol Hum Reprod 22:193-9
Zhang, Zhenbin; Sun, Liangliang; Zhu, Guijie et al. (2016) 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. Anal Chem 88:877-82