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In utero pulse injection of isotopic amino acids quantifies protein turnover rates during murine fetal development
Protein translational control is highly regulated step in the gene expression program during mammalian development that is critical for ensuring that the fetus develops correctly and that all of the necessary organs and tissues are formed and functional. Defects in protein expression during fetal de...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10245746/ https://www.ncbi.nlm.nih.gov/pubmed/37293076 http://dx.doi.org/10.1101/2023.05.18.541242 |
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author | Baeza, Josue Coons, Barbara E. Lin, Zongtao Riley, John Mendoza, Mariel Peranteau, William H. Garcia, Benjamin A |
author_facet | Baeza, Josue Coons, Barbara E. Lin, Zongtao Riley, John Mendoza, Mariel Peranteau, William H. Garcia, Benjamin A |
author_sort | Baeza, Josue |
collection | PubMed |
description | Protein translational control is highly regulated step in the gene expression program during mammalian development that is critical for ensuring that the fetus develops correctly and that all of the necessary organs and tissues are formed and functional. Defects in protein expression during fetal development can lead to severe developmental abnormalities or premature death. Currently, quantitative techniques to monitor protein synthesis rates in a developing fetus (in utero) are limited. Here, we developed a novel in utero stable isotope labeling approach to quantify tissue-specific protein dynamics of the nascent proteome during mouse fetal development. Fetuses of pregnant C57BL/6J mice were injected with isotopically labeled lysine (Lys8) and arginine (Arg10) via the vitelline vein at various gestational days. After treatment, fetal organs/tissues including brain, liver, lung, and heart were harvested for sample preparation and proteomic analysis. We show that the mean incorporation rate for injected amino acids into all organs was 17.50 ± 0.6%. By analyzing the nascent proteome, unique signatures of each tissue were identified by hierarchical clustering. In addition, the quantified proteome-wide turnover rates (k(obs)) were calculated between 3.81E-5 and 0.424 hour(−1). We observed similar protein turnover profiles for analyzed organs (e.g., liver versus brain), however, their distributions of turnover rates vary significantly. The translational kinetic profiles of developing organs displayed differentially expressed protein pathways and synthesis rates which correlated with known physiological changes during mouse development. |
format | Online Article Text |
id | pubmed-10245746 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
spelling | pubmed-102457462023-06-08 In utero pulse injection of isotopic amino acids quantifies protein turnover rates during murine fetal development Baeza, Josue Coons, Barbara E. Lin, Zongtao Riley, John Mendoza, Mariel Peranteau, William H. Garcia, Benjamin A bioRxiv Article Protein translational control is highly regulated step in the gene expression program during mammalian development that is critical for ensuring that the fetus develops correctly and that all of the necessary organs and tissues are formed and functional. Defects in protein expression during fetal development can lead to severe developmental abnormalities or premature death. Currently, quantitative techniques to monitor protein synthesis rates in a developing fetus (in utero) are limited. Here, we developed a novel in utero stable isotope labeling approach to quantify tissue-specific protein dynamics of the nascent proteome during mouse fetal development. Fetuses of pregnant C57BL/6J mice were injected with isotopically labeled lysine (Lys8) and arginine (Arg10) via the vitelline vein at various gestational days. After treatment, fetal organs/tissues including brain, liver, lung, and heart were harvested for sample preparation and proteomic analysis. We show that the mean incorporation rate for injected amino acids into all organs was 17.50 ± 0.6%. By analyzing the nascent proteome, unique signatures of each tissue were identified by hierarchical clustering. In addition, the quantified proteome-wide turnover rates (k(obs)) were calculated between 3.81E-5 and 0.424 hour(−1). We observed similar protein turnover profiles for analyzed organs (e.g., liver versus brain), however, their distributions of turnover rates vary significantly. The translational kinetic profiles of developing organs displayed differentially expressed protein pathways and synthesis rates which correlated with known physiological changes during mouse development. Cold Spring Harbor Laboratory 2023-05-21 /pmc/articles/PMC10245746/ /pubmed/37293076 http://dx.doi.org/10.1101/2023.05.18.541242 Text en https://creativecommons.org/licenses/by/4.0/This work is licensed under a Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/) , which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use. |
spellingShingle | Article Baeza, Josue Coons, Barbara E. Lin, Zongtao Riley, John Mendoza, Mariel Peranteau, William H. Garcia, Benjamin A In utero pulse injection of isotopic amino acids quantifies protein turnover rates during murine fetal development |
title | In utero pulse injection of isotopic amino acids quantifies protein turnover rates during murine fetal development |
title_full | In utero pulse injection of isotopic amino acids quantifies protein turnover rates during murine fetal development |
title_fullStr | In utero pulse injection of isotopic amino acids quantifies protein turnover rates during murine fetal development |
title_full_unstemmed | In utero pulse injection of isotopic amino acids quantifies protein turnover rates during murine fetal development |
title_short | In utero pulse injection of isotopic amino acids quantifies protein turnover rates during murine fetal development |
title_sort | in utero pulse injection of isotopic amino acids quantifies protein turnover rates during murine fetal development |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10245746/ https://www.ncbi.nlm.nih.gov/pubmed/37293076 http://dx.doi.org/10.1101/2023.05.18.541242 |
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