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Article ; Online: Isolation and Processing of Bovine Oocytes for Small RNA Sequencing.

Tan, Minjie / van Tol, Helena T A / Roovers, Elke F

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2509, Page(s) 83–91

Abstract: In modern biomedical research, mice have been the mammalian model system of choice to investigate molecular pathways for potential future medical applications. Over the last years, it has become clear that female mice employ an exceptional piRNA pathway- ... ...

Abstract	In modern biomedical research, mice have been the mammalian model system of choice to investigate molecular pathways for potential future medical applications. Over the last years, it has become clear that female mice employ an exceptional piRNA pathway-independent mechanism to neutralize transposon activity in the ovary. In other model organisms studied to date, the piRNA pathway is indispensable for efficient targeting of transposable elements and fertility in both males and females. Moreover, recent studies have demonstrated that in other mammals, including humans, the piRNA pathway is highly active in the female germline as well, indicating that the situation in the mouse female germline is anomalous. For this reason, novel models to study piRNA pathways in female mammalian germlines are currently emerging, including Bos taurus. Here we describe a protocol for isolation and downstream processing of female bovine tissues in order to perform downstream applications including piRNA sequencing.
MeSH term(s)	Animals ; Argonaute Proteins/genetics ; Cattle ; DNA Transposable Elements/genetics ; Female ; Germ Cells/metabolism ; Male ; Mammals/genetics ; Mice ; Oocytes/metabolism ; RNA, Small Interfering/metabolism ; Sequence Analysis, RNA
Chemical Substances	Argonaute Proteins ; DNA Transposable Elements ; RNA, Small Interfering
Language	English
Publishing date	2022-07-07
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2380-0_5
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Article: Cellular Fragments in the Perivitelline Space Are Not a Predictor of Expanded Blastocyst Quality.

Yu, Bo / van Tol, Helena T A / Stout, Tom A E / Roelen, Bernard A J

Frontiers in cell and developmental biology

2021 Volume 8, Page(s) 616801

Abstract: The presence of cellular fragments in the perivitelline space is a commonly used parameter to determine quality before transfer ... ...

Abstract	The presence of cellular fragments in the perivitelline space is a commonly used parameter to determine quality before transfer of
Language	English
Publishing date	2021-01-05
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2737824-X
ISSN	2296-634X
ISSN	2296-634X
DOI	10.3389/fcell.2020.616801
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Article ; Online: Reverse transcription priming methods affect normalisation choices for gene expression levels in oocytes and early embryos.

Yu, Bo / van Tol, Helena T A / Stout, Tom A E / Roelen, Bernard A J

Molecular human reproduction

2021 Volume 27, Issue 7

Abstract: Mammalian oocytes and embryos rely exclusively on maternal mRNAs to accomplish early developmental processes. Since oocytes and early embryos are transcriptionally silent after meiotic resumption, most of the synthesised maternal mRNA does not undergo ... ...

Abstract	Mammalian oocytes and embryos rely exclusively on maternal mRNAs to accomplish early developmental processes. Since oocytes and early embryos are transcriptionally silent after meiotic resumption, most of the synthesised maternal mRNA does not undergo immediate translation but is instead stored in the oocyte. Quantitative RT-PCR is commonly used to quantify mRNA levels, and correct quantification relies on reverse transcription and the choice of reference genes. Different methods for reverse transcription may affect gene expression determination in oocytes. In this study, we examined the suitability of either random or oligo(dT) primers for reverse transcription to be used for quantitative RT-PCR. We further looked for changes in poly(A) length of the maternal mRNAs during oocyte maturation. Our data indicate that depending on the method of reverse transcription, the optimal combination of reference genes for normalisation differed. Surprisingly, we observed a shortening of the poly(A) tail lengths of maternal mRNA as oocytes progressed from germinal vesicle to metaphase II. Overall, our findings suggest dynamic maternal regulation of mRNA structure and gene expression during oocyte maturation and early embryo development.
MeSH term(s)	Animals ; Blastomeres/metabolism ; Cattle ; DNA Primers/chemical synthesis ; DNA, Complementary/genetics ; Embryo Culture Techniques ; Gene Expression Regulation, Developmental ; Genes ; Morula/metabolism ; Oocytes/metabolism ; Poly A/analysis ; RNA, Messenger/genetics ; RNA, Messenger/isolation & purification ; RNA, Messenger/metabolism ; Reference Standards ; Research Embryo Creation ; Reverse Transcriptase Polymerase Chain Reaction/methods ; Reverse Transcription ; Sequence Alignment ; Sequence Homology, Nucleic Acid ; Zygote/metabolism
Chemical Substances	DNA Primers ; DNA, Complementary ; RNA, Messenger ; Poly A (24937-83-5)
Language	English
Publishing date	2021-06-20
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1324348-2
ISSN	1460-2407 ; 1360-9947
ISSN (online)	1460-2407
ISSN	1360-9947
DOI	10.1093/molehr/gaab040
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Article: Fatty Acid Supplementation During

Aardema, H / Bertijn, I / van Tol, Hta / Rijneveld, A / Vernooij, Jcm / Gadella, B M / Vos, Plam

Frontiers in cell and developmental biology

2022 Volume 10, Page(s) 837405

Abstract: ... In ... ...

Abstract	In vitro
Language	English
Publishing date	2022-03-09
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2737824-X
ISSN	2296-634X
ISSN	2296-634X
DOI	10.3389/fcell.2022.837405
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Article ; Online: Bovine

Asimaki, K / Vazakidou, P / van Tol, H T A / Oei, C H Y / Modder, E A / van Duursen, M B M / Gadella, B M

Frontiers in toxicology

2022 Volume 4, Page(s) 811285

Abstract: Endocrine disrupting chemicals (EDCs) can interfere with normal hormonal action and regulation. Exposure of women to EDCs has been associated with adverse reproductive health outcomes. The assays currently used to identify EDCs that elicit female ... ...

Abstract	Endocrine disrupting chemicals (EDCs) can interfere with normal hormonal action and regulation. Exposure of women to EDCs has been associated with adverse reproductive health outcomes. The assays currently used to identify EDCs that elicit female reproductive toxicity lack screening tests that address effects on the maturation of oocytes, a process that enables them to be fertilized and develop into embryos. Here, a screening method employing the bovine model of
Language	English
Publishing date	2022-05-24
Publishing country	Switzerland
Document type	Journal Article
ISSN	2673-3080
ISSN (online)	2673-3080
DOI	10.3389/ftox.2022.811285
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Article ; Online: An overview on how cumulus cells interact with the oocyte in a condition with elevated NEFA levels in dairy cows.

Aardema, Hilde / van Tol, Helena T A / Vos, Peter L A M

Animal reproduction science

2019 Volume 207, Page(s) 131–137

Abstract: Metabolic stress in humans and animals is associated with impaired fertility. A major characteristic of metabolic stress is elevated levels of free fatty acids (NEFAs) in blood due to mobilization of body fat reserves. Dairy cows undergo a period of ... ...

Abstract	Metabolic stress in humans and animals is associated with impaired fertility. A major characteristic of metabolic stress is elevated levels of free fatty acids (NEFAs) in blood due to mobilization of body fat reserves. Dairy cows undergo a period of metabolic stress during the peri-calving period, the so-called negative energy balance (NEB) in the early weeks postpartum. At the time of NEB, both saturated and unsaturated NEFAs are mobilized to serve as an alternative energy supply for cells, however in particular saturated NEFAs can have a detrimental effect on somatic cells. Circulating NEFAs are also reflected in the follicular fluid of ovarian follicles and hence reach the cumulus-oocyte-complex (COC), which implies a potential risk for the developing oocyte. To this end, the current review focusses on the impact of NEFAs on the quality of the oocyte.
MeSH term(s)	Animal Feed ; Animal Nutritional Physiological Phenomena ; Animals ; Cattle ; Cell Communication/physiology ; Cumulus Cells/physiology ; Dairying ; Fatty Acids, Nonesterified/blood ; Female ; Follicular Fluid/metabolism ; Lactation/metabolism ; Oocytes/physiology
Chemical Substances	Fatty Acids, Nonesterified
Language	English
Publishing date	2019-06-04
Publishing country	Netherlands
Document type	Journal Article ; Review
ZDB-ID	429674-6
ISSN	1873-2232 ; 0378-4320
ISSN (online)	1873-2232
ISSN	0378-4320
DOI	10.1016/j.anireprosci.2019.06.003
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Article ; Online: Initiation of X Chromosome Inactivation during Bovine Embryo Development.

Yu, Bo / van Tol, Helena T A / Stout, Tom A E / Roelen, Bernard A J

Cells

2020 Volume 9, Issue 4

Abstract: X-chromosome inactivation (XCI) is a developmental process that aims to equalize the dosage of X-linked gene products between XY males and XX females in eutherian mammals. In female mouse embryos, paternal XCI is initiated at the 4-cell stage; however, ... ...

Abstract	X-chromosome inactivation (XCI) is a developmental process that aims to equalize the dosage of X-linked gene products between XY males and XX females in eutherian mammals. In female mouse embryos, paternal XCI is initiated at the 4-cell stage; however, the X chromosome is reactivated in the inner cell mass cells of blastocysts, and random XCI is subsequently initiated in epiblast cells. However, recent findings show that the patterns of XCI are not conserved among mammals. In this study, we used quantitative RT-PCR and RNA in situ hybridization combined with immunofluorescence to investigate the pattern of XCI during bovine embryo development. Expression of
MeSH term(s)	Animals ; Blastocyst/metabolism ; Cattle ; DNA Methylation ; Embryonic Development/genetics ; Embryonic Development/physiology ; Gene Expression Regulation, Developmental/genetics ; Genomic Imprinting/genetics ; Histones/metabolism ; Humans ; RNA, Long Noncoding/genetics ; RNA, Long Noncoding/metabolism ; X Chromosome Inactivation/physiology
Chemical Substances	Histones ; RNA, Long Noncoding
Language	English
Publishing date	2020-04-19
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells9041016
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Article ; Online: Microinjection induces changes in the transcriptome of bovine oocytes.

Tan, Minjie / van Tol, Helena T A / Mokry, Michal / Stout, Tom A E / Roelen, Bernard A J

Scientific reports

2020 Volume 10, Issue 1, Page(s) 11211

Abstract: Gene knockdown techniques are widely used to examine the function of specific genes or proteins. While a variety of techniques are available, a technique commonly used on mammalian oocytes is mRNA knockdown by microinjection of small interfering RNA ( ... ...

Abstract	Gene knockdown techniques are widely used to examine the function of specific genes or proteins. While a variety of techniques are available, a technique commonly used on mammalian oocytes is mRNA knockdown by microinjection of small interfering RNA (siRNA), with non-specific siRNA injection used as a technical control. Here, we investigate whether and how the microinjection procedure itself affects the transcriptome of bovine oocytes. Injection of non-specific siRNA resulted in differential expression of 119 transcripts, of which 76 were down-regulated. Gene ontology analysis revealed that the differentially regulated genes were enriched in the biological processes of ATP synthesis, molecular transport and regulation of protein polyubiquitination. This study establishes a background effect of the microinjection procedure that should be borne in mind by those using microinjection to manipulate gene expression in oocytes.
MeSH term(s)	Animals ; Cattle ; Female ; Gene Expression Regulation, Developmental ; Gene Knockdown Techniques/adverse effects ; Gene Knockdown Techniques/methods ; Microinjections/adverse effects ; Oocytes/metabolism ; RNA, Messenger/genetics ; RNA, Small Interfering/administration & dosage ; RNA, Small Interfering/metabolism ; RNA-Seq ; Single-Cell Analysis ; Transcriptome/genetics
Chemical Substances	RNA, Messenger ; RNA, Small Interfering
Language	English
Publishing date	2020-07-08
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-020-67603-4
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Article ; Online: Lysophosphatidic Acid Accelerates Bovine In Vitro-Produced Blastocyst Formation through the Hippo/YAP Pathway.

Yu, Bo / van Tol, Helena T A / Oei, Christine H Y / Stout, Tom A E / Roelen, Bernard A J

International journal of molecular sciences

2021 Volume 22, Issue 11

Abstract: The segregation of trophectoderm (TE) and inner cell mass in early embryos is driven primarily by the transcription factor CDX2. The signals that trigger CDX2 activation are, however, less clear. In mouse embryos, the Hippo-YAP signaling pathway is ... ...

Abstract	The segregation of trophectoderm (TE) and inner cell mass in early embryos is driven primarily by the transcription factor CDX2. The signals that trigger CDX2 activation are, however, less clear. In mouse embryos, the Hippo-YAP signaling pathway is important for the activation of CDX2 expression; it is less clear whether this relationship is conserved in other mammals. Lysophosphatidic acid (LPA) has been reported to increase YAP levels by inhibiting its degradation. In this study, we cultured bovine embryos in the presence of LPA and examined changes in gene and protein expression. LPA was found to accelerate the onset of blastocyst formation on days 5 and 6, without changing the TE/inner cell mass ratio. We further observed that the expression of
MeSH term(s)	Acyltransferases/genetics ; Adaptor Proteins, Signal Transducing/genetics ; Animals ; Blastocyst/drug effects ; Blastocyst Inner Cell Mass/drug effects ; CDX2 Transcription Factor/genetics ; Cattle ; Cell Lineage/genetics ; Embryonic Development/drug effects ; Embryonic Development/genetics ; Gene Expression Regulation, Developmental/drug effects ; Lysophospholipids/pharmacology ; Mice ; Protein-Serine-Threonine Kinases/genetics ; Signal Transduction/drug effects ; Transcription Factors/genetics ; Trophoblasts/drug effects
Chemical Substances	Adaptor Proteins, Signal Transducing ; CDX2 Transcription Factor ; Lysophospholipids ; Transcription Factors ; Yap1 protein, mouse ; Acyltransferases (EC 2.3.-) ; tafazzin protein, mouse (EC 2.3.-) ; Hippo protein, mouse (EC 2.7.11.1) ; Protein-Serine-Threonine Kinases (EC 2.7.11.1) ; lysophosphatidic acid (PG6M3969SG)
Language	English
Publishing date	2021-05-31
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms22115915
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Article ; Online: Exposure to elevated glucose concentrations alters the metabolomic profile of bovine blastocysts.

Uhde, Karen / van Tol, Helena T A / Stout, Tom A E / Roelen, Bernard A J

PloS one

2018 Volume 13, Issue 6, Page(s) e0199310

Abstract: Chronically high blood glucose concentrations are a characteristic of diabetes mellitus. Maternal diabetes affects the metabolism of early embryos and can cause a delay in development. To mimic maternal diabetes, bovine in vitro fertilization and embryo ... ...

Abstract	Chronically high blood glucose concentrations are a characteristic of diabetes mellitus. Maternal diabetes affects the metabolism of early embryos and can cause a delay in development. To mimic maternal diabetes, bovine in vitro fertilization and embryo culture were performed in fertilization medium and culture medium containing 0.5, 2, 3, and 5 mM, glucose whereas under control conditions, the medium was glucose free (0 mM). Compared to control conditions (0 mM, 31%), blastocyst development was decreased to 23% with 0.5 and 2 mM glucose. Presence of 3 or 5 mM glucose in the medium resulted in decreased blastocyst rates (20% and 10% respectively). The metabolomic profile of resulting day 8 blastocysts was analysed by UPLC-MS/MS, and compared to that of blastocysts cultured in control conditions. Elevated glucose concentrations stimulated an increase in glycolysis and activity of the hexosamine pathway, which is involved in protein glycosylation. However, components of the tricarboxylic acid cycle, such as citrate and alpha-ketoglutarate, were reduced in glucose stimulated blastocysts, suggesting that energy production from pyruvate was inefficient. On the other hand, activity of the polyol pathway, an alternative route to energy generation, was increased. In short, cattle embryos exposed to elevated glucose concentrations during early development showed changes in their metabolomic profile consistent with the expectations of exposure to diabetic conditions.
MeSH term(s)	Animals ; Blastocyst/drug effects ; Blastocyst/metabolism ; Cattle ; Cleavage Stage, Ovum/cytology ; Cleavage Stage, Ovum/drug effects ; Culture Media, Conditioned/pharmacology ; Embryonic Development/drug effects ; Female ; Glucose/toxicity ; Metabolome/drug effects
Chemical Substances	Culture Media, Conditioned ; Glucose (IY9XDZ35W2)
Language	English
Publishing date	2018-06-20
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0199310
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