Oviductal microvesicles and their effect on in vitro maturation of canine oocytes

Consiglio, A. Lange; Perrini, C.; Albini, G.; Modina, S.; Lodde, Valentina; Orsini, E.B.; Esposti, Paola; Crémonesi, F.

doi:10.1530/rep-17-0117

Cited by 65 publications

(59 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To date, EVs have been identified in the oviduct of different species (bovine, mouse, porcine, avian, and turtle) and their functional effects have been studied in gametes and embryos (reviewed in Almiñana and Bauersachs [12]). For example, it has been shown that oviductal EVs (oEVs) support bovine embryonic development [13,14], canine oocyte maturation [15], modulate sperm capacitation and sperm fertilizing ability in the mouse and in the cat [11,16], and regulate polyspermy fertilization in the pig [17]. Regarding the effects of oEVs on embryonic development, our laboratory previously demonstrated that oEVs are taken up by the bovine embryo during in vitro culture, and that the supplementation of oEVs during in vitro embryo culture improved embryonic development and quality in terms of blastocyst rates, cell number, and hatching rates [13].…”

Section: Introductionmentioning

confidence: 99%

The Oviductal Extracellular Vesicles’ RNA Cargo Regulates the Bovine Embryonic Transcriptome

Bauersachs

Mermillod

Almiñana

2020

IJMS

View full text Add to dashboard Cite

Oviductal extracellular vesicles (oEVs) are emerging as key players in the gamete/embryo–oviduct interactions that contribute to successful pregnancy. Various positive effects of oEVs on gametes and early embryos have been found in vitro. To determine whether these effects are associated with changes of embryonic gene expression, the transcriptomes of embryos supplemented with bovine fresh (FeEVs) or frozen (FoEVs) oEVs during in vitro culture compared to controls without oEVs were analyzed by low-input RNA sequencing. Analysis of RNA-seq data revealed 221 differentially expressed genes (DEGs) between FoEV treatment and control, 67 DEGs for FeEV and FoEV treatments, and minor differences between FeEV treatment and control (28 DEGs). An integrative analysis of mRNAs and miRNAs contained in oEVs obtained in a previous study with embryonic mRNA alterations pointed to direct effects of oEV cargo on embryos (1) by increasing the concentration of delivered transcripts; (2) by translating delivered mRNAs to proteins that regulate embryonic gene expression; and (3) by oEV-derived miRNAs which downregulate embryonic mRNAs or modify gene expression in other ways. Our study provided the first high-throughput analysis of the embryonic transcriptome regulated by oEVs, increasing our knowledge on the impact of oEVs on the embryo and revealing the oEV RNA components that potentially regulate embryonic development.

show abstract

Section: Introductionmentioning

confidence: 99%

The Oviductal Extracellular Vesicles’ RNA Cargo Regulates the Bovine Embryonic Transcriptome

Bauersachs

Mermillod

Almiñana

2020

IJMS

View full text Add to dashboard Cite

show abstract

“…Optimal conditions for in vitro maturation (IVM) of canine oocytes are not yet established (Luvoni, Chigioni, Allievi, & Macis, ), although a wide variety of supplemented culture medium with different compounds, steroids, gonadotropins, oviductal microvesicles or bi‐phasic systems were tested (Apparicio et al., , ; Lange‐Consiglio et al., ; Sato et al., ). The highly variable results could be due either to suboptimal culture conditions and/or to the low competence of female gametes to develop into a fertilizable oocyte and into an embryo.…”

Section: Introductionmentioning

confidence: 99%

Nuclear competence and genetic expression of growth differentiation factor‐9 (GDF‐9) of canine oocytes in 3D culture

Morselli

Loiacono

Colombo

et al. 2018

Reprod Domestic Animals

View full text Add to dashboard Cite

To evaluate the ability of a 3D culture system in improving the nuclear and molecular competence of canine oocytes, barium alginate microcapsules were used for in vitro maturation (IVM) and the expression profile of one selected oocyte‐secreted factor, the growth differentiation factor‐9 (GDF‐9) was analysed. In Experiment I, canine grade I cumulus–oocyte complexes (COCs) were in vitro matured in 3D microcapsules in a controlled atmosphere for 72 hr, and meiosis resumption rates were compared to those of oocytes cultured in traditional 2D microdrops of medium. In Experiment II, a primer pair specific for canine GDF‐9 was designed, and preliminary tested in conventional PCR on genomic DNA. Total RNA content was isolated from oocytes at different time intervals (T0‐T24‐T48‐T72) during in vitro 3D culture, and a reverse transcription to cDNA was performed. The expression of target gene was assessed by quantitative Reverse Transcription Real‐Time PCR (qRT‐PCR), and the obtained amplicons were sequenced to check the specificity of the analysis. Canine COCs resumed meiosis at higher rates in 3D microcapsules than in 2D microdrops (p < 0.05), even though no significant differences in the proportions of oocytes achieving full maturational stages were obtained. A significant dynamic decrease in GDF‐9 expression was recorded during culture: after 72 hr of IVM, the GDF‐9 transcription significantly dropped (p = 0.018) compared to 24 and 48 hr. In conclusion, in vitro 3D culture represents an efficient system for IVM of canine oocytes, and the expression profile of GDF‐9 well reflects temporal dynamics for the acquisition of developmental competence in this species.

show abstract

“…The in vitro embryo production is a standard protocol composed of three steps: collection of oocytes and their in vitro maturation, in vitro fertilization, and in vitro culture of embryos. In our laboratory, these steps are standardized and performed according to the protocol of Perrini et al 11 and Lange-Consiglio et al 40 .…”

Section: Methodsmentioning

confidence: 99%

Amniotic microvesicles impact hatching and pregnancy percentages of in vitro bovine embryos and blastocyst microRNA expression versus in vivo controls

Consiglio

Lazzari

Pizzi

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

Embryo development and implantation are dynamic processes, responsive to external signals, and can potentially be influenced by many environmental factors. The aims of this study were to evaluate the effects of a culture medium supplemented with amniotic-derived microvesicles (MVs) on in vitro embryo hatching after cryopreservation, and pregnancy rate following embryo transfer. In addition, miRNA profiling of blastocysts produced in vitro, with or without (control; CTR) amniotic MV supplementation, was also evaluated using blastocysts produced in vivo. In vitro embryos were cultured with and without amniotic MV supplementation. In vivo blastocysts were obtained from superovulated cows. Samples for RNA isolation were obtained from three pools of 10 embryos each (in vivo, in vitro-ctR and in vitro + MVs). Our results show that the hatching percentage of cryopreserved in vitro + MVs embryos is higher (P < 0.05) than in vitro-CTR embryos and the pregnancy rate with fresh and cryopreserved in vitro + MVs embryos is higher than in vitro-CTR embryos. In addition, the analysis of differently expressed (DE) microRNAs showed that embryos produced in vivo are clearly different from those produced in vitro. Moreover, in vitro-ctR and in vitro + MVs embryos differ significantly for expression of two miRNAs that were found in higher concentrations in in vitro-CTR embryos. Interestingly, these two miRNAs were also reported in degenerated bovine embryos compared to good quality blastocysts. In conclusion, MV addition during in vitro production of embryos seems to counteract the adverse effect of in vitro culture and partially modulate the expression of specific miRNAs involved in successful embryo implantation.In the mammalian reproductive tract, the oviduct secretes a variety of growth factors and cytokines, which may play an essential role in the development of initial stages of pre-implantation embryos 1 . The absence of these maternal-embryo signals could be an important cause of the poor quality of in vitro produced embryos (IVP), compared to those collected in vivo 2,3 . To mimic the in vivo crosstalk between oviduct and embryo and to improve the quality of in vitro produced embryos, co-culture systems with somatic cells have been widely used to increase blastocyst percentage and quality of the resulting embryos, and the induction of specific transcriptomic changes 4 .Unfortunately, concern regarding viral transmission has resulted in the elimination of co-culture and the development of improved culture methods characterized by definite medium as synthetic oviductal fluid with amino acids (SOFaa 5 ). However, the maternal-embryo communication in vivo and the communication between monolayer ('helper') cells and embryos in vitro, take place not only through soluble factors (growth factors, receptors and binding proteins) secreted by cells and embryos in the medium 6,7 but also from insoluble factors. Recent studies have demonstrated that some molecules, including mRNA fragments and microRNAs (miRNAs), cannot

show abstract

Oviductal microvesicles and their effect on in vitro maturation of canine oocytes

Cited by 65 publications

References 52 publications

The Oviductal Extracellular Vesicles’ RNA Cargo Regulates the Bovine Embryonic Transcriptome

The Oviductal Extracellular Vesicles’ RNA Cargo Regulates the Bovine Embryonic Transcriptome

Nuclear competence and genetic expression of growth differentiation factor‐9 (GDF‐9) of canine oocytes in 3D culture

Amniotic microvesicles impact hatching and pregnancy percentages of in vitro bovine embryos and blastocyst microRNA expression versus in vivo controls

Contact Info

Product

Resources

About