Alfonso Gutiérrez‐Adán

Institute for Animal Reproduction

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D. Rizos

Institute for Animal Reproduction

129

Raúl Fernández‐González

Institute for Animal Reproduction

Eva Pericuesta

Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria

Belén Pintado

Universidad Autónoma de Madrid

P. Lonergan

University College Dublin

Pablo Bermejo-Ãlvarez

Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria

MÁ

Miguel Ángel Ramírez

University of Notre Dame

Isabel Gómez‐Redondo

Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria

Benjamín Planells

University of Nottingham

Priscila Ramos‐Ibeas

Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria

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Cornell University

230

University of Córdoba

228

Consejo Superior de Investigaciones Científicas

126

Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria

120

Centro Regional de Selección y Reproducción Animal

Instituto de Salud Carlos III

Institute for Research and Technology in Food and Agriculture

Inserm

Centro de Investigación Biomédica en Red

Universidad Complutense de Madrid

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Additional file 5: Table S4. of Suboptimal culture conditions induce more deviations in gene expression in male than female bovine blastocysts

Figshare (2016)

Sonia Heras Dieter De Coninck Mario Van Poucke Karen Goossens Osvaldo Bogado Pascottini

GO, in terms of the biological process, molecular function, and cellular component, and KEGG pathways of the genes differentially expressed (FDR corrected p-value <0.05, â FCâ â Ľ2) in all the comparisons performed. (XLSX 66 kb)

Table (database)

10.6084/m9.figshare.c.3632816_d5

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Effects of Synchronous and Asynchronous Embryo Transfer on Postnatal Development, Adult Health, and Behavior in Mice1

Biology of Reproduction (2015)

Angela Patricia López‐Cardona Raúl Fernández‐González Míriam Pérez-Crespo Francisco Alén Fernando Rodrı́guez de Fonseca

Asynchronous embryo transfer (ET) is a common assisted reproduction technique used in several species, but its biological effects on postnatal and early development remain unknown. The aim of this study was to determine whether asynchronous ET produces long-term effects in mice. Postnatal development, animal weight, systolic blood pressure (SBP), relative organ weight (liver, spleen, kidneys, heart, lungs, brain, and testicles), and behavior (assessed in open-field and elevated plus maze tests) were assessed in CD1 mice produced by different ET procedures: 1) the transfer of Day 3.5 (D3.5) blastocysts to the uterus (BL-UT); 2) the transfer of D3.5 blastocysts to the oviduct (BL-OV); or 3) the transfer of D0.5 zygotes to the oviduct (Z-OV). In vivo conceived animals served as controls (CT). The transfer of blastocysts to the uterus or zygotes to the oviduct was defined as synchronous, and transfer of blastocysts to the oviduct was defined as asynchronous. Both synchronous and asynchronous ET resulted in increased weight at birth that normalized thereafter with the exception of asynchronous ET females. In this group, female BL-OV, a clear lower body weight was recorded along postnatal life when compared with controls (P < 0.05). No effects on animal weight were produced during postnatal development in the synchronous ET groups (BL-UT, Z-OV, and CT). Both synchronous and asynchronous ET had impacts on adult (Wk 30) organ weight. SBP was modified in animals derived from blastocyst but not zygote ET. Effects on behavior (anxiety in the plus maze) were only detected in the BL-UT group (P < 0.05). Our findings indicate that zygotes are less sensitive than blastocysts to ET and that both synchronous and asynchronous blastocyst ET may have long-term consequences on health, with possible impacts on weight, arterial pressure, relative organ weight, and behavior.

Oviduct

10.1095/biolreprod.115.130385

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Citations (15)

Author response for "A high glucose concentration during early stages of <i>in vitro</i> equine embryo development alters expression of genes involved in glucose metabolism"

María Jesús Sánchez‐Calabuig Raúl Fernández‐González Meriem Hamdi Katrien Smits Angela Patricia López‐Cardona

Carbohydrate Metabolism

10.1111/evj.13342/v5/response1

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Author response for "A high glucose concentration during early stages of <i>in vitro</i> equine embryo development alters expression of genes involved in glucose metabolism"

María Jesús Sánchez‐Calabuig Raúl Fernández‐González Meriem Hamdi Katrien Smits Angela Patricia López‐Cardona

Carbohydrate Metabolism

10.1111/evj.13342/v3/response1

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Semen quality at cryobanking before oncologic treatment

Yasuyo Matsumoto Lévai Írisz Sakae Goto Hidehisa HASHIMOTO S. Kokeguchi

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Fathers Produced by In Vitro Culture Program Obesity and Glucose Intolerance in Male Mice Offspring.

Biology of Reproduction (2011)

Alexandra Calle Raúl Fernández‐González Alberto Miranda Bedate Ricardo Laguna Alfonso Gutiérrez‐Adán

10.1093/biolreprod/85.s1.202

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Effect of long-term culture of mouse embryonic stem cells under low oxygen concentration as well as on glycosaminoglycan hyaluronan on cell proliferation and differentiation

Cell Proliferation (2010)

Miguel Ángel Ramírez Eva Pericuesta María Yáñez‐Mó Artur Pałasz Alfonso Gutiérrez‐Adán

Objectives: Maintaining undifferentiated stem cells in defined conditions is of critical importance to improve their in vitro culture.We have evaluated the effects of culturing mouse stem (mES) cells under physiological oxygen concentration as well as by replacing fibroblast feeder layer (mEF) with gelatin or glycosaminoglycan hyaluronan (HA), on cell proliferation and differentiation.Materials and methods: After 3 days culture or after long-term cell culture under different conditions, levels of apoptotic cell death were determined by cell cycle and TUNEL (TdT-mediated dUTP nick end labelling) assays and levels of cell proliferation by CFSE (5-(and-6)-carboxyfluorescein diacetate succinimidyl ester) labelling.We assessed spontaneous differentiation into cardiomyocytes and mRNA expression of pluripotency and differentiation biomarkers.Results: After 3 days culture under hypoxic conditions, levels of proliferation and apoptosis of mES cells were higher, in correlation with increase in intracellular reactive oxygen species.However, when cells were continuously grown for 1 month under those conditions, the level of apoptosis was, in all cases, under 4%.Hypoxia reduced spontaneous differentiation of mES into cardiomyocytes.Long-term culture on HA was more effective in maintaining the pluripotent state of the mES cells when compared to that on gelatin.Level of terminal differentiation was highest on mEF, intermediate on HA and lowest on gelatin.Conclusions: Our data suggest that hypoxia is not necessary for maintaining pluripotency of mES cells and appeared to be detrimental during ES differentiation.Moreover, HA may offer a valuable alternative for long-term culture of mES cells in vitro.

10.1111/j.1365-2184.2010.00732.x

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Mouse ICSI with frozen–thawed sperm: The impact of sperm freezing procedure and sperm donor strain

Molecular Reproduction and Development (2003)

Pedro N. Moreira Adela Jiménez Raúl Fernández‐González Ninoska Bury‐Madrid Julio de la Fuente

Normal mouse offspring can be obtained from oocytes injected with frozen-thawed spermatozoa without cryoprotection, however, embryo development can be affected by sperm freezing procedure and sperm donor strain. In this study we observed that direct contact of mouse spermatozoa with liquid nitrogen did not affect their ability to activate injected oocytes but severely restricted subsequent in vitro embryo development to blastocyst stage. Tris-EDTA buffer and M2 were also shown to be better sperm freezing extenders than DPBS, allowing higher developmental potential. In addition, differences in embryo development obtained by intracytoplasmic sperm injection (ICSI) with frozen-thawed spermatozoa were observed between hybrid sperm donor strains. Frozen-thawed B6D2F1 spermatozoa provided higher embryo development than sperm cells from C57CBAF1.

10.1002/mrd.10335

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235RETINOID-DEPENDENT MRNA EXPRESSION IN BOVINE OOCYTES PREMATURED AND/OR MATURED IN VITRO

Reproduction Fertility and Development (2004)

E. Gómez C. Dı́ez E. Morán Alejandro Ávalos‐Rodríguez L. J. Royo

As a transcription factor, retinoic acid (RA) can activate or silence a wide number of genes, thus inducing differentiation in cell systems and playing a role in cell cycle regulation. However, little is known of RA-dependent gene expression in the oocyte. Bovine oocytes and cumulus cells express most RA receptors, and the presence of 9-cis-RA during in vitro maturation (IVM) is beneficial to oocyte development (Duque et al., 2002 Hum. Reprod. 17, 2706–2714; Hidalgo et al., 2003 Reproduction 125, 409–416). The present work analyzes the relative abundance of various developmentally important gene transcripts in bovine oocytes during in vitro prematuration and/or maturation. Cumulus-oocyte complexes (COCs) were manipulated in defined medium with polyvinyl-alcohol (DM-PVA). Those COCs undergoing prematuration were cultured for 24h in DM-PVA with 25μM roscovitine. For IVM, some prematured COCs were cultured for 24h in DM-PVA containing pFSH, LH and E2. Incubations were made at 39°C in an atmosphere of 5% CO2 in air and high humidity. Within experiments, COCs were cultured with nM 9-cis-RA 5, in 1% ethanol (both as vehicle and inhibitor of endogenous RA synthesis), 3% ethanol, 5% ethanol and untreated. Using Real Time PCR (10 oocytes per group) (Rizos et al., 2003 Biol. Reprod. 68, 236) we examined the relative mRNA expression of genes involved in protection against free oxygen radicals (Mn-superoxide dismutase, MnSOD), glucose metabolism (glucose-6-phosphate dehydrogenase, G6PDH) and cell cycle events (Cyclin B1 and H1). Data (of 4 replicates) were analyzed by ANOVA and Duncan test (P<0.05). Regarding immature oocytes, prematuration in 1% ethanol increased cyclin B1 expression and decreased cyclin H1, while 9-cis-RA increased G6PDH. Maturation without additives increased cyclin B1 and G6PDH, but decreased cyclin H1 and MnSOD expression;; opposite trends were observed under increasing ethanol dosages (3% and 5%). Maturation with 1% ethanol or 9-cis-RA enhanced cyclin B1 and G6PDH, while reducing cyclin H1 and MnSOD expressions. The presence of 9-cis-RA during both prematuration and maturation processes tended to show more prominent effects than the ones observed when it was present only during prematuration or maturation alone. In our study, in presence of 9-cis-RA during both prematuration and maturation processes, the expression of cyclin B1 and G6PDH tended to increase, while cyclin H1 and MnSOD tended to decrease. However, the differences with the control group without additives were not significant. Our study during both prematuration and maturation processes show that beneficial effects of RA on oocyte developmental competence may not be related to the alteration of mRNA expression of the four genes analyzed. Grant support: Spanish Ministry of Science and Technology (AGL-2002-01175; 2003-05783).

In vitro maturation

Growth differentiation factor-9

10.1071/rdv16n1ab235

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New Challenges in the Analysis of Gene Transcription in Bovine Blastocysts

Reproduction in Domestic Animals (2011)

Pablo Bermejo-Ãlvarez Eva Pericuesta Alberto Miranda Bedate C. de Frutos Serafín Pérez‐Cerezales

Contents In the last years, enormous progress has been made in the analysis of gene transcription at the blastocyst stage. The study of gene expression at this early stage of development is challenging because of the very small amount of starting material, which limits the use of traditional mRNA analysis approaches such as Northern blot. Another problem is the difficulty for data normalization, particularly the identification of the best housekeeping gene with the lowest fluctuation under different developmental conditions. Moreover, the transcriptional analysis of embryo biopsies or individual embryos needs to take into consideration that the blastocyst is a transitional stage of development, which is composed of three different types of cells (trophoblast, epiblast and primitive ectoderm) with different patterns of gene expression, and that there are large differences between male and female blastocysts. In this review, we analyse the different specific and sensitive tools available to compare mRNA expression levels of specific genes at the blastocyst stage, and how the protocol and the analytical method used can influence the results dramatically. Finally, we describe future research challenges to identify candidate genes related to developmental competence of bovine blastocysts, not only in terms of pregnancy rates but also in relation to adverse long‐term consequences in the adult animal.

Housekeeping gene

Trophoblast

Epiblast

Northern blot

10.1111/j.1439-0531.2011.01842.x

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Citations (7)