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Vertebrate reproductive science and technology
RESEARCH ARTICLE

Temporally differential protein expression of glycolytic and glycogenic enzymes during in vitro preimplantation bovine embryo development

Manuel García-Herreros A B C , Constantine A. Simintiras B and Patrick Lonergan B
+ Author Affiliations
- Author Affiliations

A National Institute for Agricultural and Veterinary Research (INIAV, I.P.), Quinta da Fonte Boa 2005-048, Santarém, Portugal.

B School of Agriculture and Food Science, University College Dublin, Belfield, Dublin 4, Dublin D04 N2E5, Ireland.

C Corresponding author. Email: herrerosgm@gmail.com

Reproduction, Fertility and Development 30(9) 1245-1252 https://doi.org/10.1071/RD17429
Submitted: 17 October 2017  Accepted: 1 March 2018   Published: 23 March 2018

Abstract

Proteomic analyses are useful for understanding the metabolic pathways governing embryo development. This study investigated the presence of enzymes involved in glycolysis and glycogenesis in in vitro-produced bovine embryos at five developmental stages leading up to blastocyst formation. The enzymes examined were: (1) glycolytic: hexokinase-I (HK-I), phosphofructokinase-1 (PFK-1), pyruvate kinase mutase 1/2 (PKM-1/2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and (2) glycogenic: glycogen synthase kinase-3 isoforms α/ β (GSK-3α/β). Glucose transporter-1 (GLUT-1) was also analysed. The developmental stages examined were: (1) 2–4-cell, (2) 5–8-cell, (3) 16-cell, (4) morula and (5) expanded blastocyst. The enzymes HK-I, PFK-1, PKM-1/2, GAPDH and GLUT-1 were differentially expressed throughout all stages (P < 0.05). GSK-3α and β were also differentially expressed from the 2–4-cell to the expanded blastocyst stage (P < 0.05) and GLUT-1 was identified throughout. The general trend was that the abundance of PFK1, GAPDH and PKM-1/2 decreased whereas HK-I, phospho-GSK3α (P-GSK3α) and P-GSK3β levels increased as the embryo advanced. In contrast, GLUT-1 expression peaked at the 16-cell stage. These data combined suggest that in vitro bovine embryo metabolism switches from being glycolytic-centric to glycogenic-centric around the 16-cell stage, the developmental window also characterised by embryonic genome activation.

Additional keywords: cattle, developmental stages, early embryos, metabolic pathways, signal transduction.


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