1,721,226 research outputs found
Experimental data on in vitro sperm capacitation in presence of cAMP and MK571
No full textcAMP has been reported to be an essential driver of sperm capacitation and cAMP efflux through multidrug resistance-associated protein 4 (MRP4) has been suggested to maintain intracellular cAMP homeostasis and generate extracellular signaling able to regulate capacitation. Using an in vitro model of pig sperm capacitation, this study investigated the influence of extracellular cAMP on sperm capacitation and acquisition of fertilizing ability, and evaluated the role of MRP4 using MK571, a MRP4 selective inhibitor.
These data sets contain the raw data on sperm tyrosine-phosphorylated protein localization, sperm viability and in vitro fertilization (IVF) output
Experimental data about mammalian sperm function and survival. Effects of Roundup and its main component, glyphosate
No full textThe wide use of glyphosate-based herbicides (GBHs) has become a matter of concern due to its potential harmful effects on human health, including men fertility. The study investigated, using the pig as a model, the impact of pure glyphosate and its most known commercial formulation, Roundup, on sperm function and survival.
These data sets contain the raw data of the flow cytometry analysis of sperm parameters (sperm viability, acrosome integrity, mitochondrial activity and DNA fragmentation) and motility analysis by computer-assisted sperm analysis (CASA) of fresh commercial semen doses incubated with different concentrations (0–360 μg/mL) of glyphosate (GLY) or Roundup (R), at the equivalent GLY concentration for 3 h
Experimental Data on In Vitro Maturation of Cumulus-Enclosed Oocites Exposed to Polystyrene Nanoplastics
No full textIn recent years, there is increasing concern about the potential harmful effects of nanoplastics (NPs) on mammalian reproductive processes, especially regarding their impact on gametes. Given the limited research on NPs effect on mammalian oocyte, this study investigated the biological impact of increasing concentrations of 100 nm polystyrene nanoplastics (PS-NPs)(5, 50, 100 and 200 μg/mL) on cumulus-enclosed oocytes (COCs), using an in vitro model of pig oocyte maturation (IVM).
These data sets contain the raw data on nuclear maturation, cytoplasmic maturation and developmental competence of oocytes, intracellular oocyte levels of glutathione (GSH) and reactive oxygen species (ROS), and steroidogenic activity of cumulus cells (CCs)
Is Resveratrol Effective in Protecting Stallion Cooled Semen?
No full textThe aim of this work was to evaluate the effect of resveratrol (RSV) during liquid storage of
stallion sperm for 24 hours at either 10C or 4C. The antioxidant RSV was added to reduce
the oxidative damage that occurs during cold storage. Aliquots of 2 mL of diluted semen
were stored either at 4C or 10C under anaerobic conditions, in the absence (control
group) or presence of RSV at different concentrations (10, 20, 40, and 80 mM). Sperm
quality parameters were assessed at 0 hours and after 24 hours of storage. Resveratrol
treatment did not affect sperm quality parameters at 0 hours. At 24-hour storage, a significant
(P < .01) decrease of sperm quality was observed independently from RSV supplementation
and storage temperature. A significant decrease of viable spermatozoa with
high mitochondrial membrane potential (SYBRþ/PI/JC-1þ) was evident at 24-hour
storage in 40- and 80-mM RSV groups compared with control group. Moreover, a decline
of total motility in 80-mM RSV group compared with the control group and a decrease of
progressive motility and average path velocity in 80-mM RSV group compared with control
and 20-mM RSV groups were observed. In conclusion, our findings demonstrate that RSV
supplementation does not enhance sperm quality of stallion semen after 24 hours of
storage. Moreover, 40- and 80-mM RSV concentrations could damage sperm functional
status, probably acting as pro-oxidant. Finally, although 24-hour storage significantly
affected most of the sperm quality parameters, no significant differences were found in
groups maintained at 4C or 10C, suggesting that stallion semen could be equally preserved
at these different temperatures
EFFECT OF STAINING AND SORTING ON BOAR SPERM MEMBRANE INTEGRITY, MITOCHONDRIAL ACTIVITY AND IN VITRO BLASTOCYST DEVELOPMENT
No full textThe objective of this study was to determine the effects of staining with Hoechst 33342 and of the entire sorting procedure on boar sperm membrane integrity (using Annexin-V/PI), mitochondrial activity (using JC-1/SYBR/PI) and blastocyst development in vitro; the effect of storage at 17 degrees C for 24h prior to Hoechst staining and sorting was also investigated. The Hoechst staining and the whole sorting procedure reduced the percent of live spermatozoa in both fresh (day 0) and stored (day 1) semen, as determined by both assays; nevertheless, there was no increase in live sperm cells showing signs of early damage (Annexin-V positive, propidium negative), whose percentages remained nearly zero. The majority of Annexin-V positive cells were propidium positive, therefore dead. JC-1 staining evidenced a correlation between mitochondrial activity and viability. However, a significant difference between viable sperm cells and sperm cells with active mitochondria was detected in control and stained sperm, whereas almost all viable sorted spermatozoa had active mitochondria. No significant differences in the in vitro produced blastocysts both on day 0 and 1 were observed. In conclusion, despite the damages induced by sorting procedures, semen sorted as fresh or after storage at 17 degrees C can be successfully used for in vitro production of pig embryos
Porcine circovirus type 2 detection in in vitro produced porcine blastocysts after virus sperm exposure
Full text linkThis study was aimed at assessing the capability of semen experimentally infected with porcine circovirus type 2 (PCV2) to produce porcine blastocysts PCR positive for PCV2. Embryos were obtained from in vitro maturation (IVM) and in vitro fertilization (IVF) of porcine oocytes or by parthenogenesis. Sperm suspension was exposed to PCV2b and utilized for IVF. PCV2 spiked semen did not reveal any reduction in sperm viability or motility but its ability to produce infected blastocysts was irrelevant as only one out of 15 blastocysts obtained by IVF were PCV2b; however two blastocysts were PCV2a positive. Furthermore, the presence of PCV2 was demonstrated also in embryos obtained by parthenogenesis (one out of 17 was PCV2b and one PCV2a positive). Even if PCV2 firmly attaches to the surface of spermatozoa, experimentally spiked sperm were not effective in infecting oocytes during IVF and in producing PCR positive embryos. The infected blastocysts we obtained derived most probably from infected oocytes recovered at the abattoir
Sperm function and mitochondrial activity: An insight on boar sperm metabolism
Full text linkIn this study boar sperm mitochondrial activity was studied and deepened in order to delineate the main
metabolic strategies used by boar sperm to obtain energy and to link them to sperm function.
Boar spermatozoa were collected, diluted at 30 106 spz/mL and incubated for 1 h with: Rotenone
(ROT), complex I inhibitor, Dimethyl-malonate (DMM), complex II inhibitor, antimycin A (ANTI), complex
III inhibitor, oligomycin (OLIGO), ATP synthase inhibitor, Carbonyl cyanide m-chlorophenyl hydrazone
(CCCP), uncoupling agent, 2-deoxy-glucose (2DG), glucose agonist, and Dimethyl sulphoxide (DMSO) as
control vehicle. Viability and mitochondrial membrane potential (Sybr14/PI/JC1 staining) and sperm
motility (using CASA system) were assayed after incubation.
ROT, ANTI, OLIGO and CCCP significantly reduced total and progressive motility as well as cell velocities;
ANTI and CCCP depressed mitochondrial membrane potential but did not affect cell viability.
Cluster analysis of kinematic parameters showed some interesting features of sperm subpopulations:
ANTI and CCCP caused a shift in sperm subpopulation towards “slow non progressive” cells, OLIGO and
ROT caused a shift towards “average” and “slow non progressive” cells, while DMM and 2DG increased
the “fast progressive” cells subpopulation.
Sperm mitochondrial respiration and substrate oxidation, assayed polographically and spectrofluorimetrically,
respectively pointed out a high ATP turnover and a low spare respiratory capacity, mainly
linked to the NADH-O2 oxidase activity. Therefore, boar spermatozoa heavily rely on mitochondrial
oxidative phosphorylation, and especially on Complex I activity, to produce ATP and fuel motility
X and Y chromosome-bearing spermatozoa are equally able to uptake and internalize exogenous DNA by sperm-mediated gene transfer in swine
No full textSince proteomic differences between male X/Y chromosome-bearing gametes have recently been described, a question has been raised: could these differences be responsible for different behavior between X and Y chromosome-bearing spermatozoa during the binding and internalization of exogenous DNA in the swine species? In order to investigate this hypothesis, our group studied the process of the uptake and internalization of exogenous DNA in X and Y chromosome-bearing sperm sub-populations. No significant differences were found between sperm types in both the uptake and internalization of exogenous DNA. The quantity of internalized exogenous DNA was significantly lower than that of the uptaken DNA. In conclusion, our results showed that X and Y chromosomes-bearing spermatozoa have the same binding capacity and internalization of DNA, and the proteomic differences between them do not seem to interfere with these complex processe
Involvement of extracellular vesicle-encapsulated miRNAs in human reproductive disorders: a systematic review
Full text linkIn recent years, extracellular vesicles (EVs) have emerged as essential players in cell-to-cell communication, particularly having an active regulating role in biological systems. Because reproductive-associated processes are not exempt of this communication, multiple studies have been devoted to this realm, focusing on gamete maturation, embryo implantation or fetal development. The aim of the present review was to comprehensively and systematically collect evidence about the function of the microRNA (miRNA) encapsulated in EVs isolated from different reproductive tissues or fluids in reproductive-related diseases. Following PRISMA guidelines, we conducted a systematic search of the literature published in MEDLINE-PubMed until the end of February 2021. After selection, 32 studies were included in the qualitative review comparing the miRNA expression profile in EVs between different pathological disorders. Most reports showed the potential of the miRNAs carried by EVs to be used as putative biomarkers of reproductive disorders, including pregnancy affections, disease progression and quality of preimplantation embryos. The most relevant miRNAs were found to be highly heterogeneous among studies, with some conflicting results. Further research is thus warranted to address whether cofounding factors, such as the methods to isolate EVs and miRNAs, the subset of EVs, the criteria of patient selection, the timing of sample retrieval, or any other factor, may explain the inconsistencies between studies
Combining reduced glutathione and ascorbic acid has supplementary beneficial effects on boar sperm cryotolerance
No full textThe main aim of this work was to evaluate how supplementing freezing and thawing media with reduced glutathione (GSH) and l-ascorbic acid (AA) affected the quality parameters of frozen-thawed boar spermatozoa. With this purpose, semen samples of 12 ejaculates coming from 12 boars were used. Each ejaculate was split into seven aliquots to which 5 mM of GSH and 100 μM of AA were added separately or together at two different steps of freeze-thawing. Various sperm parameters (levels of free cysteine residues in sperm nucleoproteins, sperm viability, acrosome membrane integrity, intracellular peroxide and superoxide levels [ROS], and total and progressive motility) were evaluated before freezing and at 30 and 240 minutes after thawing. Both GSH and AA significantly improved boar sperm cryotolerance when they were separately added to freezing and thawing media. However, the highest improvement was recorded when both freezing and thawing media were supplemented with 5 mM of GSH plus 100 μM of AA. This improvement was observed in sperm viability and acrosome integrity, sperm motility, and nucleoprotein structure. Although ROS levels were not much increased by freeze-thawing procedures, the addition of GSH and AA to both freezing and thawing extenders significantly decreased intracellular peroxide levels and had no impact on superoxide levels. According to our results, we can conclude that supplementation of freezing and thawing media with both GSH and AA has a combined, beneficial effect on frozen-thawed boar sperm, which is greater than that obtained with the separate addition of either GSH or AA
- …
