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Disrupting porcine glutaminase does not block preimplantation development and elongation nor decrease mTORC1

Paula R Chen1, Caroline G Lucas1, Raissa F Cecil1

  • 1Division of Animal Sciences, University of Missouri, Columbia, MO, USA.

Biology of Reproduction
|August 28, 2021
PubMed
Summary
This summary is machine-generated.

Porcine glutaminase (GLS) is not essential for pig conceptus elongation or mechanistic target of rapamycin complex 1 (mTORC1) activation. GLS knockout embryos elongated, indicating GLS is not required for this crucial developmental process.

Keywords:
elongationglutaminemechanistic target of rapamycin complex 1porcinepreimplantation development

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Area of Science:

  • Reproductive Biology
  • Developmental Biology
  • Biochemistry

Background:

  • Conceptus elongation in pigs is vital for pregnancy establishment and relies on nutrient supply.
  • Glutamine is a key energy substrate and influences mechanistic target of rapamycin complex 1 (mTORC1) signaling during early embryonic development.
  • The role of glutamine metabolism, specifically glutaminase (GLS), beyond the blastocyst stage in pigs remains understudied.

Purpose of the Study:

  • To investigate the necessity of glutaminase (GLS) for conceptus elongation in pigs.
  • To determine if GLS is involved in activating mTORC1 signaling during porcine conceptus elongation.
  • To elucidate the metabolic and signaling pathways influencing conceptus development.

Main Methods:

  • CRISPR/Cas9 gene editing was employed to create GLS knockout (GLS-/-) porcine fetal fibroblasts.
  • Somatic cell nuclear transfer (SCNT) was used to generate GLS+/+ and GLS-/- embryos.
  • Embryos were transferred to surrogate mothers, and conceptuses were collected on day 14 of gestation for morphological and molecular analysis.

Main Results:

  • GLS-/- conceptuses exhibited diverse morphologies (spherical, ovoid, tubular, filamentous) and successfully elongated, despite absent GLS protein and minimal enzyme activity.
  • Spherical GLS-/- conceptuses showed increased transcripts for glutamine and glutamate metabolism and transport compared to filamentous conceptuses.
  • No significant differences in mTORC1 component phosphorylation were observed between genotypes or morphologies, though targets like cyclin D1 were elevated in spherical conceptuses.

Conclusions:

  • Porcine glutaminase (GLS) is not essential for conceptus elongation past the blastocyst stage.
  • GLS is not required for mTORC1 activation during porcine conceptus elongation.
  • Alternative metabolic pathways likely support conceptus elongation in the absence of functional GLS.