Improving porcine in vitro fertilization output by simulating the oviductal environment

Soriano‐Úbeda, Cristina; García–Vázquez, Francisco Alberto; Romero-Aguirregomezcorta, Jon; Matás, Carmen

doi:10.1038/srep43616

Cited by 19 publications

(22 citation statements)

References 63 publications

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“…COCs were collected from antral follicles (3‐6 mm diameter), and only those with complete and dense cumulus oophorus were used for IVM. COCs were separated in groups of 50 and cultured for 42 hours in 500 µL NCSU‐37 medium following the protocol described by Funahashi et al, CM was obtained as described by Soriano‐Úbeda et al, The NCSU‐37 IVM medium from wells where groups of 50 COCs had completed the second phase of IVM (with FF but without dbAMPc, PMSG, and hCG) was pipetted to mix COCs’ secretions with the surrounding medium. After that, the medium was centrifuged at 7000 g for 10 minutes at 4ºC and the pellet containing cellular debris was discarded.…”

Section: Methodsmentioning

confidence: 99%

“…Research suggests that some molecules present in the OF as well as in the cumulus oophorus are involved in the modulation of sperm function and their acquisition of fertilizing capacity . The OF is a product of both serum transudate and active secretion from the epithelium and is composed of numerous metabolites, enzymes, growth factors, hormones, proteases, simple and complex carbohydrates, ions, lipids, phospholipids, glycosidases, glycosyltransferases, glycosaminoglycans, proteoglycans, chaperones, and heat shock proteins .…”

Section: Introductionmentioning

confidence: 99%

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Periovulatory oviductal fluid decreases sperm protein kinase A activity, tyrosine phosphorylation, and in vitro fertilization in pig

et al. 2020

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Background: Molecules from the female reproductive tract modulate capacitation and function of sperm cells in vivo. These molecules vary in a quantitative and qualitative manner throughout the estrous cycle. Objectives: This work evaluates the effect of using various female reproductive fluids on capacitation and fertilization of pig spermatozoa in vitro. Material and methods: The effects of culturing spermatozoa in different fluids on the levels of sperm protein kinase A (pPKA), tyrosine phosphorylation, acrosome reaction, and in vitro fertilization (IVF) were evaluated. The fluids tested were as follows: oviductal fluid (OF) from five phases of the estrous cycle, namely early and late follicular (OF-EF, OF-LF), early and late luteal (OF-EL, OF-LL) and periovulatory (pOF), follicular fluid from medium-sized follicles, and secretions of cumulus-oocyte complexes (conditioned medium). Results:The pPKAs and tyrosine phosphorylation were decreased by OF-EF, OF-LF, OF-EL, and pOF but not by follicular fluid and conditioned medium. OF-EF, OF-LF, and pOF also decreased the sperm acrosome reaction. Moreover, the effect of pOF on pPKAs and tyrosine phosphorylation was reversible. In in vitro fertilization, OF-EF, OF-LF, OF-EL, and pOF reduced the percentage of penetrated oocytes, the mean number of spermatozoa per penetrated oocyte, and increased monospermy.Conclusion: OF from follicular, early luteal, and periovulatory phases of the estrous cycle modulates the sperm protein phosphorylation as well as the acrosome reaction involved in capacitation and increases monospermic fertilization in in vitro fertilization. Our findings suggest that fluids from the female reproductive tract could be used as additives in porcine IVF systems to modulate sperm-oocyte interaction. K E Y W O R D Sboar, fluid, in vitro fertilization, oviduct, capacitation

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Periovulatory oviductal fluid decreases sperm protein kinase A activity, tyrosine phosphorylation, and in vitro fertilization in pig

et al. 2020

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“…In vivo, key reproductive processes, including gamete maturation and fertilization occur within the oviductal fluid (ODF) . With assisted reproductive technology (ART), the physiological situation has been imitated in vitro . Thus, ODF has been recognized in playing a critical role in the modulation of the reproductive processes, by influencing the viability and motility of spermatozoa, oocyte maturation, gamete interaction, and early embryo development .…”

Section: Introductionmentioning

confidence: 99%

“…With assisted reproductive technology (ART), the physiological situation has been imitated in vitro. [2,3] Thus, ODF has been recognized in playing a critical role in the modulation of the reproductive processes, by influencing the viability and motility of spermatozoa, oocyte maturation, gamete interaction, and early embryo development. [1,[4][5][6][7] Moreover, ODF affects the capacitation of spermatozoa in vitro.…”

Section: Introductionmentioning

confidence: 99%

Identification of Rabbit Oviductal Fluid Proteins Involved in Pre‐Fertilization Processes by Quantitative Proteomics

Hackenbroch

Meyer

et al. 2019

Proteomics

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Oviductal fluid (ODF) proteins modulate and support reproductive processes in the oviduct. In the present study, proteins involved in the biological events that precede fertilization have been identified in the rabbit ODF proteome, isolated from the ampulla and isthmus of the oviduct at different time points within 8 h after intrauterine insemination. A workflow is used that integrates lectin affinity capture with stable‐isotope dimethyl labeling prior to nanoLC‐MS/MS analysis. In total, over 400 ODF proteins, including 214 lectin enriched glycoproteins, are identified and quantified. Selected data are validated by Western blot analysis. Spatiotemporal alterations in the abundance of ODF proteins in response to insemination are detected by global analysis. A subset of 63 potentially biologically relevant ODF proteins is identified, including extracellular matrix components, chaperones, oxidoreductases, and immunity proteins. Functional enrichment analysis reveals an altered peptidase regulator activity upon insemination. In addition to protein identification and abundance changes, N‐glycopeptide analysis further identifies 281 glycosites on 199 proteins. Taken together, these results show, for the first time, the evolving oviductal milieu early upon insemination. The identified proteins are likely those that modulate in vitro processes, including spermatozoa function.

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“…However, despite some clear evidences found in vitro, little attention has been paid to mimicking the in vivo sperm transit, specially through the uterine horns, for further use in ARTs. Thus, pH, CO 2 or HCO 3 − variations in culture media during in vitro gamete preparation and/or interaction showed relevant consequences in the fertilization output [49,50]. Also, an in uence of the uterine uid composition on sperm selection and consequent embryo culture was observed in vitro in several species including porcine [51][52][53].…”

Section: Discussionmentioning

confidence: 99%

In vivo measurement of pH, CO2 and HCO3- levels in the uterus of non-anesthetized sows through the estrous cycle and after insemination

López-Albors¹,

Llamas-López²,

Ortuño³

et al. 2020

Preprint

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Background The pH-CO2-HCO3− system is a ubiquitous biological regulator with important functional implications for reproduction. Knowledge of the physiological values of its components is relevant for reproductive biology and the optimization of Assistant Reproductive Techniques (ARTs). In vivo pH of the oviduct and uterus has been estimated by direct in situ measurements in a few species. However, regarding the levels of CO2 and HCO3−, information is very scarce and, when available, it comes from fluid samples instead of in vivo estimations. This study describes a non-invasive method to measure pH and % of CO2 in the uterus of sows with cutting-edge technology and no medication. Sows were at three different reproductive conditions, estrous with no insemination E(-)AI and after insemination E(+)AI, and diestrous (non-estrous, NE). From pH and CO2 data, HCO3− concentration was estimated. Results The designed methodology allowed for in situ time-lapse recording of pH and % of CO2 within the uterus of non-anesthetized sows. Variable oscillatory patterns of pH, CO2 and HCO3− were found independently of the estrous condition. Insemination did not changed the levels of uterine pH, % of CO2 and HCO3− concentration, -E(-)AI = E(+)AI-, but all the values were affected by the estrous cycle in a way that decreased significantly at diestrous condition - E(-)AI and E(+)AI > NE-. Conclusions A non-invasive approach to the porcine uterus with novel optical probes allowed the obtaining of in situ physiological values of pH, CO2, and HCO3− at different reproductive conditions. While the short-time presence of sperm in the uterus did not change the physiological milieu, the whole pH-CO2-HCO3− system was affected by the estrous cycle. This study contributes to a better understanding of the in vivo regulation of the pH/CO2/HCO3− system in the uterus and may help to optimize the protocols of sperm treatment for in vitro fertilization.

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Improving porcine in vitro fertilization output by simulating the oviductal environment

Cited by 19 publications

References 63 publications

Periovulatory oviductal fluid decreases sperm protein kinase A activity, tyrosine phosphorylation, and in vitro fertilization in pig

Periovulatory oviductal fluid decreases sperm protein kinase A activity, tyrosine phosphorylation, and in vitro fertilization in pig

Identification of Rabbit Oviductal Fluid Proteins Involved in Pre‐Fertilization Processes by Quantitative Proteomics

In vivo measurement of pH, CO2 and HCO3- levels in the uterus of non-anesthetized sows through the estrous cycle and after insemination

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