Glucose and redox metabolism in meiotically blocked in vitro grown mouse antral follicles

Anamaria-Cristina Herta; Lucia von Mengden; Nazli Akin; Katy Billooye; Wim Coucke; Berta Cava-Cami; Fábio Klamt; Johan Smitz; Ellen Anckaert

doi:10.1007/s10815-023-02940-7

Glucose and redox metabolism in meiotically blocked in vitro grown mouse antral follicles

Anamaria-Cristina Herta, Lucia von Mengden, Nazli Akin, Katy Billooye, Wim Coucke, Berta Cava-Cami, Fábio Klamt, Johan Smitz, Ellen Anckaert

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Abstract

PURPOSE: Glucose and redox metabolism characterization in mouse antral follicles with meiotically blocked oocytes, after in vitro follicle culture (IFC) from the early secondary stage.

METHODS: Following IFC (10 days), oocytes, corresponding cumulus (CC), and granulosa cells (GC) were collected from antral follicles: (i) on day 9-immature, germinal vesicle (GV) stage; (ii) on day 10, after hCG/EGF stimulation-mature, metaphase II (MII) stage and meiotically blocked (MB) immature GV stage. The metabolic profiles of all samples (GV, MII, and MB) were compared by measuring changes in metabolites involved in glycolysis, tricarboxylic acid (TCA) cycle, pentose phosphate pathway (PPP), and redox activity via enzymatic spectrophotometric assays in each cell type.

RESULTS: Within MB follicles, GCs drive higher levels of glycolysis and lactic acid fermentation (LAF) while oocytes exert more PPP activity. MB-oocytes had significantly larger diameters compared to day 9 GVs. MB follicles revealed limited metabolic changes in the somatic compartment compared to their GV counterparts (before stimulation). MB-CCs showed increased aconitase and glucose-6-phosphate dehydrogenase activities with lower malate levels comparted to GV-CCs. MB and MII in vitro grown follicles displayed comparable metabolic profiles, suggesting culture induces metabolic exhaustion regardless of the maturation stage.

CONCLUSIONS: Current results suggest that in addition to impaired nuclear maturation, metabolic disruption is present in MB follicles. MB follicles either compensate with high levels of TCA cycle and PPP activities in CCs, or are unable to drive proper levels of aerobic metabolism, which might be due to the current culture conditions.

Original language	English
Pages (from-to)	2851-2863
Number of pages	13
Journal	Journal of Assisted Reproduction and Genetics
Volume	40
Issue number	12
DOIs	https://doi.org/10.1007/s10815-023-02940-7
Publication status	Published - Dec 2023

Bibliographical note

Funding Information:
This study was funded by Flanders FWO The Excellence of Science (EOS, FWO- F.R.S.-FNRS; G0F3118N) and by the Brazilian funds MCTI (Ministério da Ciência, Tecnologia e Inovações)/CNPq (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico) INCTTM (Instituto Nacional de Ciência e Tecnologia Translacional em Medicina)/CAPES (Coordenaçao de Aperfeiçoamento de Pessoal de Nivel Superior)/FAPESP (465458/2014-9) and PDSE-CAPES (Programa de Doutorado Sanduiche no Exterior-Coordenaçao de Aperfeiçoamento de Pessoal de Nivel Superior) 47/2017 (88881.188914/2018-01).

Publisher Copyright:
© 2023, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

Cumulus cells
Failed oocyte maturation
Glucose metabolism
Granulosa cells
Mouse secondary follicle culture

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Cite this

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title = "Glucose and redox metabolism in meiotically blocked in vitro grown mouse antral follicles",

abstract = "PURPOSE: Glucose and redox metabolism characterization in mouse antral follicles with meiotically blocked oocytes, after in vitro follicle culture (IFC) from the early secondary stage.METHODS: Following IFC (10 days), oocytes, corresponding cumulus (CC), and granulosa cells (GC) were collected from antral follicles: (i) on day 9-immature, germinal vesicle (GV) stage; (ii) on day 10, after hCG/EGF stimulation-mature, metaphase II (MII) stage and meiotically blocked (MB) immature GV stage. The metabolic profiles of all samples (GV, MII, and MB) were compared by measuring changes in metabolites involved in glycolysis, tricarboxylic acid (TCA) cycle, pentose phosphate pathway (PPP), and redox activity via enzymatic spectrophotometric assays in each cell type.RESULTS: Within MB follicles, GCs drive higher levels of glycolysis and lactic acid fermentation (LAF) while oocytes exert more PPP activity. MB-oocytes had significantly larger diameters compared to day 9 GVs. MB follicles revealed limited metabolic changes in the somatic compartment compared to their GV counterparts (before stimulation). MB-CCs showed increased aconitase and glucose-6-phosphate dehydrogenase activities with lower malate levels comparted to GV-CCs. MB and MII in vitro grown follicles displayed comparable metabolic profiles, suggesting culture induces metabolic exhaustion regardless of the maturation stage.CONCLUSIONS: Current results suggest that in addition to impaired nuclear maturation, metabolic disruption is present in MB follicles. MB follicles either compensate with high levels of TCA cycle and PPP activities in CCs, or are unable to drive proper levels of aerobic metabolism, which might be due to the current culture conditions.",

keywords = "Cumulus cells, Failed oocyte maturation, Glucose metabolism, Granulosa cells, Mouse secondary follicle culture",

author = "Anamaria-Cristina Herta and {von Mengden}, Lucia and Nazli Akin and Katy Billooye and Wim Coucke and Berta Cava-Cami and F{\'a}bio Klamt and Johan Smitz and Ellen Anckaert",

note = "Funding Information: This study was funded by Flanders FWO The Excellence of Science (EOS, FWO- F.R.S.-FNRS; G0F3118N) and by the Brazilian funds MCTI (Minist{\'e}rio da Ci{\^e}ncia, Tecnologia e Inova{\c c}{\~o}es)/CNPq (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico) INCTTM (Instituto Nacional de Ci{\^e}ncia e Tecnologia Translacional em Medicina)/CAPES (Coordena{\c c}ao de Aperfei{\c c}oamento de Pessoal de Nivel Superior)/FAPESP (465458/2014-9) and PDSE-CAPES (Programa de Doutorado Sanduiche no Exterior-Coordena{\c c}ao de Aperfei{\c c}oamento de Pessoal de Nivel Superior) 47/2017 (88881.188914/2018-01). Publisher Copyright: {\textcopyright} 2023, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.",

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doi = "10.1007/s10815-023-02940-7",

language = "English",

volume = "40",

pages = "2851--2863",

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Glucose and redox metabolism in meiotically blocked in vitro grown mouse antral follicles. / Herta, Anamaria-Cristina; von Mengden, Lucia; Akin, Nazli ; Billooye, Katy; Coucke, Wim; Cava-Cami, Berta; Klamt, Fábio; Smitz, Johan ; Anckaert, Ellen.

In: Journal of Assisted Reproduction and Genetics, Vol. 40, No. 12, 12.2023, p. 2851-2863.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Glucose and redox metabolism in meiotically blocked in vitro grown mouse antral follicles

AU - Herta, Anamaria-Cristina

AU - von Mengden, Lucia

AU - Akin, Nazli

AU - Billooye, Katy

AU - Coucke, Wim

AU - Cava-Cami, Berta

AU - Klamt, Fábio

AU - Smitz, Johan

AU - Anckaert, Ellen

N1 - Funding Information: This study was funded by Flanders FWO The Excellence of Science (EOS, FWO- F.R.S.-FNRS; G0F3118N) and by the Brazilian funds MCTI (Ministério da Ciência, Tecnologia e Inovações)/CNPq (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico) INCTTM (Instituto Nacional de Ciência e Tecnologia Translacional em Medicina)/CAPES (Coordenaçao de Aperfeiçoamento de Pessoal de Nivel Superior)/FAPESP (465458/2014-9) and PDSE-CAPES (Programa de Doutorado Sanduiche no Exterior-Coordenaçao de Aperfeiçoamento de Pessoal de Nivel Superior) 47/2017 (88881.188914/2018-01). Publisher Copyright: © 2023, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.

PY - 2023/12

Y1 - 2023/12

N2 - PURPOSE: Glucose and redox metabolism characterization in mouse antral follicles with meiotically blocked oocytes, after in vitro follicle culture (IFC) from the early secondary stage.METHODS: Following IFC (10 days), oocytes, corresponding cumulus (CC), and granulosa cells (GC) were collected from antral follicles: (i) on day 9-immature, germinal vesicle (GV) stage; (ii) on day 10, after hCG/EGF stimulation-mature, metaphase II (MII) stage and meiotically blocked (MB) immature GV stage. The metabolic profiles of all samples (GV, MII, and MB) were compared by measuring changes in metabolites involved in glycolysis, tricarboxylic acid (TCA) cycle, pentose phosphate pathway (PPP), and redox activity via enzymatic spectrophotometric assays in each cell type.RESULTS: Within MB follicles, GCs drive higher levels of glycolysis and lactic acid fermentation (LAF) while oocytes exert more PPP activity. MB-oocytes had significantly larger diameters compared to day 9 GVs. MB follicles revealed limited metabolic changes in the somatic compartment compared to their GV counterparts (before stimulation). MB-CCs showed increased aconitase and glucose-6-phosphate dehydrogenase activities with lower malate levels comparted to GV-CCs. MB and MII in vitro grown follicles displayed comparable metabolic profiles, suggesting culture induces metabolic exhaustion regardless of the maturation stage.CONCLUSIONS: Current results suggest that in addition to impaired nuclear maturation, metabolic disruption is present in MB follicles. MB follicles either compensate with high levels of TCA cycle and PPP activities in CCs, or are unable to drive proper levels of aerobic metabolism, which might be due to the current culture conditions.

AB - PURPOSE: Glucose and redox metabolism characterization in mouse antral follicles with meiotically blocked oocytes, after in vitro follicle culture (IFC) from the early secondary stage.METHODS: Following IFC (10 days), oocytes, corresponding cumulus (CC), and granulosa cells (GC) were collected from antral follicles: (i) on day 9-immature, germinal vesicle (GV) stage; (ii) on day 10, after hCG/EGF stimulation-mature, metaphase II (MII) stage and meiotically blocked (MB) immature GV stage. The metabolic profiles of all samples (GV, MII, and MB) were compared by measuring changes in metabolites involved in glycolysis, tricarboxylic acid (TCA) cycle, pentose phosphate pathway (PPP), and redox activity via enzymatic spectrophotometric assays in each cell type.RESULTS: Within MB follicles, GCs drive higher levels of glycolysis and lactic acid fermentation (LAF) while oocytes exert more PPP activity. MB-oocytes had significantly larger diameters compared to day 9 GVs. MB follicles revealed limited metabolic changes in the somatic compartment compared to their GV counterparts (before stimulation). MB-CCs showed increased aconitase and glucose-6-phosphate dehydrogenase activities with lower malate levels comparted to GV-CCs. MB and MII in vitro grown follicles displayed comparable metabolic profiles, suggesting culture induces metabolic exhaustion regardless of the maturation stage.CONCLUSIONS: Current results suggest that in addition to impaired nuclear maturation, metabolic disruption is present in MB follicles. MB follicles either compensate with high levels of TCA cycle and PPP activities in CCs, or are unable to drive proper levels of aerobic metabolism, which might be due to the current culture conditions.

KW - Cumulus cells

KW - Failed oocyte maturation

KW - Glucose metabolism

KW - Granulosa cells

KW - Mouse secondary follicle culture

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U2 - 10.1007/s10815-023-02940-7

DO - 10.1007/s10815-023-02940-7

M3 - Article

C2 - 37776437

VL - 40

SP - 2851

EP - 2863

JO - Journal of Assisted Reproduction and Genetics

JF - Journal of Assisted Reproduction and Genetics

SN - 1058-0468

IS - 12

ER -

Glucose and redox metabolism in meiotically blocked in vitro grown mouse antral follicles

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Keywords

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