Dataset Information

Manipulation of bicarbonate concentration in sperm capacitation media improvesin vitro fertilisation output in porcine species.

ABSTRACT:

Background

The in vivo concentration of bicarbonate (HCO₃ ^-), one of the essential sperm capacitating effectors, varies greatly in the different environments sperm go through from cauda epididymis to the fertilisation site. On the contrary, porcine in vitro sperm capacitation and fertilisation media usually contains a standard concentration of 25?mmol/L, and one of the main problems presented is the unacceptable high incidence of polyspermy. This work hypothesised that by modifying the HCO₃ ^- concentration of the medium, the output of in vitro sperm capacitation and fertilisation could be increased.

Results

Once exposed to the capacitation medium, the intracellular pH (pH_i) of spermatozoa increased immediately even at low concentrations of HCO₃ ^-, but only extracellular concentrations of and above 15?mmol/L increased the substrates protein kinase A phosphorylation (pPKAs). Although with a significant delay, 15?mmol/L of HCO₃ ^- stimulated sperm linear motility and increased other late events in capacitation such as tyrosine phosphorylation (Tyr-P) to levels similar to those obtained with 25?mmol/L. This information allowed the establishment of a new in vitro fertilisation (IVF) system based on the optimization of HCO₃ ^- concentration to 15?mmol/L, which led to a 25.3% increment of the viable zygotes (8.6% in the standard system vs. 33.9%).

Conclusions

Optimising HCO₃ ^- concentrations allows for establishing an IVF method that significantly reduced porcine polyspermy and increased the production of viable zygotes. A concentration of 15?mmol/L of HCO₃ ^- in the medium is sufficient to trigger the in vitro sperm capacitation and increase the fertilisation efficiency in porcine.

SUBMITTER: Soriano-Ubeda C

PROVIDER: S-EPMC6410524 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Manipulation of bicarbonate concentration in sperm capacitation media improvesin vitro fertilisation output in porcine species.

Soriano-Úbeda Cristina C Romero-Aguirregomezcorta Jon J Matás Carmen C Visconti Pablo E PE García-Vázquez Francisco A FA

Journal of animal science and biotechnology 20190311

<h4>Background</h4>The in vivo concentration of bicarbonate (HCO<sub>3</sub> <sup>-</sup>), one of the essential sperm capacitating effectors, varies greatly in the different environments sperm go through from cauda epididymis to the fertilisation site. On the contrary, porcine in vitro sperm capacitation and fertilisation media usually contains a standard concentration of 25 mmol/L, and one of the main problems presented is the unacceptable high incidence of polyspermy. This work hypothesised t ...[more]

PMID: 30899459

Similar Datasets

Project description:The red wolf is a critically endangered canid, with ~250 and ~20 individuals in the ex situ and reintroduced wild populations, respectively. Assisted reproductive technologies such as sperm cryopreservation and in vitro fertilization therefore represent critically-needed tools to manage these populations. However, the motility of post-thaw red wolf sperm rapidly declines during in vitro incubation, hindering the ability to develop these technologies. In this study, we evaluated the influence of several culture media (a modified canine capacitation medium (mCCM), a modified North Carolina State University-23 medium (mNCSU-23), a synthetic oviductal fluid (SOF), a fertilization Tyrode's medium base or Fert-TALP (FERT), and a TRIS-based buffer (TRIS)) on the survival and capacitation of red wolf sperm during extended (18 h) incubation at 38.5°C and 5% CO2. Red wolf sperm motility averaged (±s.e.m.) 73.8 ± 7.1% at the time of collection, and was better maintained over 4 h incubation in mCCM (55.0 ± 9.8%) and mNCSU-23 (54.7 ± 10.4), compared to mSOF (43.8 ± 8.3%), FERT (30 ± 10.5), and TRIS (16.4 ± 4.1%) solutions. Patterns of tyrosine phosphorylation signal, as assessed via immunocytochemistry, indicated induction of capacitation between 2 and 4 h in vitro culture. Tyrosine phosphorylation signal was particularly robust in mCCM and mNCSU-23 incubated sperm, although significant acrosome exocytosis was not observed in response to progesterone supplementation after 3 h incubation in any of the media. In sum, results indicate mCCM and mNCSU-23 are promising base media for the in vitro incubation and capacitation of red wolf sperm, for assisted reproduction applications.Lay summaryDevelopment of assisted reproductive technologies such as in vitro fertilization and artificial insemination is of high importance to the genetic management of critically endangered species such as the red wolf (Canis rufus). However, these technologies require the ability to maintain sperm viability and function during extended incubation, which has not been successful for the red wolf thus far. In this study, various culture media developed for sperm/egg/embryo culture in large mammalian species were evaluated for their ability to maintain red wolf sperm motility under physiological incubation conditions. Media and conditions previously utilized for domestic dog sperm were found to best support sperm incubation and capacitation (process of becoming competent to fertilize an egg) in the red wolf, representing a key step for future development of assisted reproductive technologies for the species.

Project description:The male gamete is not completely mature after ejaculation and requires further events in the female genital tract to acquire fertilizing ability, including the processes of capacitation and acrosome reaction. In order to shed light on protein changes experienced by the sperm cell in preparation for fertilization, a comprehensive quantitative proteomic profiling based on isotopic peptide labeling and liquid chromatography followed by tandem mass spectrometry was performed on spermatozoa from three donors of proven fertility under three sequential conditions: purification with density gradient centrifugation, incubation with capacitation medium, and induction of acrosome reaction by exposure to the calcium ionophore A23187. After applying strict selection criteria for peptide quantification and for statistical analyses, 36 proteins with significant changes in their relative abundance within sperm protein extracts were detected. Moreover, the presence of peptide residues potentially harboring sites for post-translational modification was revealed, suggesting that protein modification may be an important mechanism in sperm maturation. In this regard, increased levels of proteins mainly involved in motility and signaling, both regulated by protein modifiers, were detected in sperm lysates following incubation with capacitation medium. In contrast, less abundant proteins in acrosome-reacted cell lysates did not contain potentially modifiable residues, suggesting the possibility that all those proteins might be relocated or released during the process. Protein-protein interaction analysis revealed a subset of proteins potentially involved in sperm maturation, including the proteins Erlin-2 (ERLIN2), Gamma-glutamyl hydrolase (GGH) and Transmembrane emp24 domain-containing protein 10 (TMED10). These results contribute to the current knowledge of the molecular basis of human fertilization. It should now be possible to further validate the potential role of the detected altered proteins as modulators of male infertility.

Dataset Information

Manipulation of bicarbonate concentration in sperm capacitation media improvesin vitro fertilisation output in porcine species.

Background

Results

Conclusions

Publications

Manipulation of bicarbonate concentration in sperm capacitation media improvesin vitro fertilisation output in porcine species.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets