Retinoids during the in vitro transition from bovine morula to blastocyst

Rodríguez, Aida; Díez, C.; Ikeda, Shuntaro; Royo, Luis J.; Caamaño, J. N.; Alonso-Montes, Cristina; Álvarez, Isabel; Facal, N.; Gómez, E.

doi:10.1093/humrep/del099

Cited by 26 publications

(38 citation statements)

References 59 publications

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“…In the present study, ATRA increased the development of Day 8 blastocysts compared with LG treatment and control, although significant differences between ATRA and LG treatment groups were already evident on Day 7. Blastocyst development on Day 7 after ATRA treatment did not differ from that recorded without additives, similar to that obtained in the presence of serum with morulae produced in B2 + Vero cells (Rodríguez et al 2006). However, in a recent paper (Rodríguez et al 2007), blastocyst rates obtained after 48 h treatment of morulae with ATRA-containing medium were similar to those obtained in the control (no additives) group, but lower than those seen in embryos cultured with 0.1 µm LG.…”

Section: Discussionsupporting

confidence: 72%

“…The wide band filters for blue and ultraviolet excitation were used for detection of TUNEL and bisbenzimide staining, respectively. Discrimination between nuclei showing apoptotic or necrotic morphology was based on a previous description (Gjørret et al 2003), as shown in Rodríguez et al (2006). Cells were regarded as apoptotic when they showed both TUNEL labelling and apoptotic morphology.…”

Section: Apoptosis and Necrosismentioning

confidence: 99%

“…However, we have demonstrated that long exposure (48 h) to 0.1 µm LG improves blastocyst development (Rodríguez et al 2007) and that short exposure (24 h) to 0.7 µm ATRA increases cell numbers in the inner cell mass (ICM) and trophectoderm (TE) without changing the ICM/TE cell ratio (Rodríguez et al 2006). Classically, the quality of the IVP bovine blastocyst has been associated with the amount and proper allocation of embryonic cells to the ICM and TE (Van Soom et al 1996), as well as their sensitivity to cryopreservation (Imai et al 2002;Lonergan et al 2003aLonergan et al , 2003bRizos et al 2003).…”

Section: Introductionmentioning

confidence: 99%

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127 Development and Quality of Bovine Morulae Cultured in Serum-Free Medium With Retinoic Receptor Specific Agonists

Gómez¹,

Caamaño²,

Muñoz³

et al. 2008

Reprod. Fertil. Dev.

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Abstract.Retinoids regulate development and differentiation of the bovine blastocyst in vitro, although the underlying mechanisms remain to be clarified. A challenge in reproductive biotechnology is the identification of pathways that regulate early embryonic development and their influence on blastocyst differentiation, apoptosis and survival to cryopreservation as traits of embryo quality. The present paper analyses the effects of short-term exposure (24 h) to retinoids on in vitroproduced bovine morulae. Immature cumulus oocyte complexes were in vitro matured and fertilised. Presumptive zygotes were subsequently cultured in modified synthetic oviduct fluid up to Day 6, in which morulae were randomly allocated to the different experimental groups. The treatments consisted of 0.1 µm LG100268 (LG; a retinoid X receptor agonist), 0.7 µm all-trans retinoic acid (ATRA; a retinoic acid receptor agonist) or no additives. Day 8 blastocyst development was increased in the ATRA-treated group compared with the LG and untreated embryos. In Day 7 embryos, the number of total cells and cells allocated to the trophectoderm were higher in the ATRA-treated group compared with untreated embryos. Apoptosis in the inner cell mass increased after LG treatment, whereas ATRA had no effect. After vitrification and warming, survival and hatching rates of Day 7 blastocysts did not change with retinoid treatment. Within the LGtreated and untreated blastocyst groups, survival and hatching rates were higher for Day 7 than Day 8 embryos; however, Day 8 blastocysts treated with ATRA showed improved hatching rates. In conclusion, treatment of morulae with ATRA in serum-free medium improves embryo development and quality without increasing the incidence of apoptosis and necrosis.

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Section: Discussionsupporting

confidence: 72%

Section: Apoptosis and Necrosismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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127 Development and Quality of Bovine Morulae Cultured in Serum-Free Medium With Retinoic Receptor Specific Agonists

Gómez¹,

Caamaño²,

Muñoz³

et al. 2008

Reprod. Fertil. Dev.

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“…Bovine embryos were in vitro produced from slaughterhouse ovaries as previously reported [15]. Presumptive zygotes were cultured in synthetic oviduc fluid (SOF) with 6 g/l BSA at 38.7 8C, 5% CO 2 , 5% O 2 and 90% N 2 .…”

Section: In Vitro Embryo Productionmentioning

confidence: 99%

“…PCR was performed with an i-Cycler iQ Real-Time PCR detection System (Bio-Rad, Hercules, CA, USA) as previously reported [15] Primer sequences, PCR traits for target genes, annealing temperature, sizes of the amplified fragments, and GeneBank accession number are shown in Table 1. Data were normalized to histone H2a1 transcript levels [16].…”

Section: Real-time Pcrmentioning

confidence: 99%

Tyrosine kinase A, C and fibroblast growth factor-2 receptors in bovine embryos cultured in vitro

et al. 2009

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Neurotrophins and basic fibroblast growth factor are ligands of tyrosine kinase receptors, though they bind to different tyrosine kinase receptor classes. Neurotrophins bind to receptor tyrosine kinase class VII, Trk receptor family, while basic fibroblast growth factor binds to receptor tyrosine kinase class IV, FGF receptor family. The mammalian uterine tract immunolocalizes neurotrophins and bFGF; therefore their cognate receptors might exert a role during embryonic development. Using RT-PCR, we found mRNA for p75 NTR TrkA, TrkC and FGFr2 throughout the early bovine embryonic development in vitro. Immunofluorescent staining, assessed by confocal microscopy, showed the expression of TrkA and TrkC proteins in oocytes and all embryonic stages analyzed. We have provided a novel description of TrkA and TrkC proteins, and TrkA, TrkC, p75 NTR and FGFr2 mRNA expression throughout mammalian embryonic development. This work may help to design future research with neurotrophins in bovine embryo culture and embryonic stem cells.

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A Single Cell Transcriptomic Fingerprint of Stressed Premature, Imbalanced Differentiation of Embryonic Stem Cells

L Ruden,

Singh,

Marben

et al. 2024

Birth Defects Research

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BackgroundMiscarriages cause a greater loss‐of‐life than cardiovascular diseases, but knowledge about environmentally induced miscarriages is limited. Cultured naïve pluripotent embryonic stem cells (ESC) differentiate into extra‐embryonic endoderm/extraembryonic endoderm (XEN) or formative pluripotent ESC, during the period emulating maximal miscarriage of peri‐implantation development. In previous reports using small marker sets, hyperosmotic sorbitol, or retinoic acid (RA) decreased naïve pluripotency and increased XEN by FACS quantitation.MethodsBulk and single cell (sc)RNAseq analyses of two cultured ESC lines was done, corroborated by qPCR. Transcriptomic responses were analyzed of cultured ESC stressed by Sorbitol, with Leukemia inhibitory factor (LIF + ; stemness growth factor), RA without LIF to control for XEN induction, and compared with normal differentiation (LIF − , ND).ResultsSorbitol and RA increase subpopulations of 2‐cell embryo‐like (2CEL) and XEN sub‐lineages; primitive, parietal, and visceral endoderm (VE) cells and suppress formative pluripotency, imbalancing alternate lineage choices of initial naïve pluripotent cultured ESC compared with ND. Although bulk RNAseq and gene ontology (GO) group analyses suggest that stress induces anterior VE‐head organizer and placental markers, scRNAseq reveals relatively few cells. But VE and placental markers/cells were in adjacent stressed cell clusters in the UMAP, like recent, normal UMAP of conceptuses. UMAPs show that dose‐dependent stress overrides stemness to force premature lineage imbalance.ConclusionsHyperosmotic stress, and other toxicological stresses, like drugs with active ingredient RA, may cause premature, lineage imbalance, resulting in miscarriages or birth defects.

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Retinoids during the in vitro transition from bovine morula to blastocyst

Cited by 26 publications

References 59 publications

127 Development and Quality of Bovine Morulae Cultured in Serum-Free Medium With Retinoic Receptor Specific Agonists

127 Development and Quality of Bovine Morulae Cultured in Serum-Free Medium With Retinoic Receptor Specific Agonists

Tyrosine kinase A, C and fibroblast growth factor-2 receptors in bovine embryos cultured in vitro

A Single Cell Transcriptomic Fingerprint of Stressed Premature, Imbalanced Differentiation of Embryonic Stem Cells

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