Display options
Cite
Chen PR, Lucas CG, Cecil RF, et al. Disrupting porcine glutaminase does not block preimplantation development and elongation nor decrease mTORC1 activation in conceptuses†. Biol Reprod. 2021;105(5):1104-1113doi: 10.1093/biolre/ioab165.
Chen, P. R., Lucas, C. G., Cecil, R. F., Pfeiffer, C. A., Fudge, M. A., Samuel, M. S., Zigo, M., Seo, H., Spate, L. D., Whitworth, K. M., Sutovsky, P., Johnson, G. A., Wells, K. D., Geisert, R. D., & Prather, R. S. (2021). Disrupting porcine glutaminase does not block preimplantation development and elongation nor decrease mTORC1 activation in conceptuses†. Biology of reproduction, 105(5), 1104-1113. https://doi.org/10.1093/biolre/ioab165
Chen, Paula R, et al. "Disrupting porcine glutaminase does not block preimplantation development and elongation nor decrease mTORC1 activation in conceptuses†." Biology of reproduction vol. 105,5 (2021): 1104-1113. doi: https://doi.org/10.1093/biolre/ioab165
Chen PR, Lucas CG, Cecil RF, Pfeiffer CA, Fudge MA, Samuel MS, Zigo M, Seo H, Spate LD, Whitworth KM, Sutovsky P, Johnson GA, Wells KD, Geisert RD, Prather RS. Disrupting porcine glutaminase does not block preimplantation development and elongation nor decrease mTORC1 activation in conceptuses†. Biol Reprod. 2021 Nov 15;105(5):1104-1113. doi: 10.1093/biolre/ioab165. PMID: 34453429.
Copy Download .nbib Format: NLM
Share it on

Biol Reprod. 2021 Nov 15;105(5):1104-1113. doi: 10.1093/biolre/ioab165.

Disrupting porcine glutaminase does not block preimplantation development and elongation nor decrease mTORC1 activation in conceptuses†.

Biology of reproduction

Paula R Chen, Caroline G Lucas, Raissa F Cecil, Caroline A Pfeiffer, Melissa A Fudge, Melissa S Samuel, Michal Zigo, Heewon Seo, Lee D Spate, Kristin M Whitworth, Peter Sutovsky, Gregory A Johnson, Kevin D Wells, Rodney D Geisert, Randall S Prather

Affiliations

  1. Division of Animal Sciences, University of Missouri, Columbia, MO, USA.
  2. Department of Veterinary Integrative Biosciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA.

PMID: 34453429 DOI: 10.1093/biolre/ioab165

Abstract

Elongation of pig conceptuses is a dynamic process, requiring adequate nutrient provisions. Glutamine is used as an energy substrate and is involved in the activation of mechanistic target of rapamycin complex 1 (mTORC1) during porcine preimplantation development. However, the roles of glutamine have not been extensively studied past the blastocyst stage. Therefore, the objective of the current study was to determine if glutaminase (GLS), which is the rate-limiting enzyme in glutamine metabolism, was necessary for conceptus elongation to proceed and was involved in mTORC1 activation. The CRISPR/Cas9 system was used to induce loss-of-function mutations in the GLS gene of porcine fetal fibroblasts. Wild type (GLS+/+) and knockout (GLS-/-) fibroblasts were used as donor cells for somatic cell nuclear transfer, and GLS+/+ and GLS-/- blastocyst-stage embryos were transferred into surrogates. On day 14 of gestation, GLS+/+ conceptuses primarily demonstrated filamentous morphologies, and GLS-/- conceptuses exhibited spherical, ovoid, tubular, and filamentous morphologies. Thus, GLS-/- embryos were able to elongate despite the absence of GLS protein and minimal enzyme activity. Furthermore, spherical GLS-/- conceptuses had increased abundance of transcripts related to glutamine and glutamate metabolism and transport compared to filamentous conceptuses of either genotype. Differences in phosphorylation of mTORC1 components and targets were not detected regarding conceptus genotype or morphology, but abundance of two transcriptional targets of mTORC1, cyclin D1, and peroxisome proliferator-activated receptor gamma coactivator 1-alpha was increased in spherical conceptuses. Therefore, porcine GLS is not essential for conceptus elongation and is not required for mTORC1 activation at this developmental timepoint.

© The Author(s) 2021. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: [email protected].

Keywords: elongation; glutamine; mechanistic target of rapamycin complex 1; porcine; preimplantation development

Publication Types

Grant support