| 1. |
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| 2. |
- Alvarez-Rodríguez, Manuel, et al.
(författare)
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The antioxidant effects of soybean lecithin- or low-density lipoprotein-based extenders for the cryopreservation of brown-bear (Ursus arctos) spermatozoa.
- 2013
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Ingår i: Reproduction, Fertility and Development. - : CSIRO Publishing. - 1031-3613 .- 1448-5990. ; 25:8, s. 1185-1193
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Tidskriftsartikel (refereegranskat)abstract
- Egg yolk low-density lipoproteins (LDL) and soybean lecithin were evaluated as replacements for egg yolk in extenders used for the cryopreservation of brown-bear spermatozoa. The motility, viability and acrosomal status of post-thawed spermatozoa were analysed, and an egg-yolk extender was used as a control. The total antioxidant capacity of these extenders was tested. Soybean lecithin showed an effect that was dependent on the soybean concentration (2%, 3.5% and 5%) and source (Type A: 24% L-α-phosphatidylcholine, and Type B: 14-23% L-α-phosphatidylcholine). Only semen cryopreserved with 5% Type A soybean exhibited a sperm motility similar to that of semen cryopreserved in egg-yolk-based extender after thawing, although the sperm viability and acrosome status were not as high. Semen frozen in an extender containing LDL (10-15%) exhibited improved sperm viability in comparison with the control, but sperm motility was lower. The LDL-based extender exhibited a higher anti-oxidant activity than the egg-yolk extender and soy lecithin-based extenders. The extenders with higher anti-oxidant activity showed improvements in frozen sperm viability but lower semen motility. These results indicate that soybean lecithin did not have the same protective effect as egg yolk during the freezing of brown-bear spermatozoa but suggest that LDL (10-15%) could be a useful substitute for egg yolk in these extenders.
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| 3. |
- Chankeaw, Wiruntita, et al.
(författare)
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Elevated non-esterified fatty acids impair survival and promote lipid accumulation and pro-inflammatory cytokine production in bovine endometrial epithelial cells
- 2018
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 30, s. 1770-1784
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Tidskriftsartikel (refereegranskat)abstract
- Elevated non-esterified fatty acids (NEFAs) are associated with negative effects on bovine theca, granulosa and oviductal cells but the effects of NEFAs on bovine endometrial epithelial cells (bEECs) are not as well documented. The objective of this study was to define the effects of NEFAs on bEECs. Postprimary bEECs were treated with 150, 300 or 500 mu M of either palmitic acid (PA), stearic acid (SA) or oleic acid (OA) or a mixture of NEFAs (150 mu M of each FA) or 0.5% final concentration of vehicle ethanol (control). Viability and proliferation of bEECs exposed to 150 mu M of each NEFA or a mixture of NEFAs were unaffected. Increased lipid accumulation was found in all treated groups (P < 0.01). In cells exposed to 500 mu M of each NEFA and 300 mu M PA decreased cell viability (P < 0.001), proliferation (P < 0.05) and increased apoptosis (P < 0.05) were observed. Treatment with 500 mu M OA, PA and SA had the strongest effects on cell viability, proliferation and apoptosis (P < 0.05). Treatment with PA and OA increased interleukin-6 (IL-6) concentrations (P < 0.05), whereas only the highest concentration of PA, OA and SA stimulated IL-8 production (P < 0.05). These results suggest that high concentrations of NEFAs may impair endometrial function with more or less pronounced effects depending on the type of NEFA and time of exposure.
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| 4. |
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| 5. |
- Cuello, C., et al.
(författare)
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Superfine open pulled straws vitrification of porcine blastocysts does not require pretreatment with cytochalasin B and/or centrifugation
- 2010
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Ingår i: Reproduction, Fertility and Development. - : CSIRO Publishing. - 1031-3613 .- 1448-5990. ; 22:5, s. 808-817
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Tidskriftsartikel (refereegranskat)abstract
- The present study investigated the in vitro development of and cytoskeletal disruption suffered by in vivo-derived porcine blastocysts subjected to superfine open pulled straws (SOPS) vitrification. Blastocysts were either untreated prior to SOPS vitrification or were subjected to one of the following three pretreatment protocols: (1) centrifugation (12 min, 13 000g); (2) 25 min equilibration with 7.5 mu g mL(-1) cytochalasin B; or (3) equilibration with cytochalasin B followed by centrifugation. After 24 h culture, fresh (n = 32) and vitrified-warmed (n = 188) blastocysts were evaluated by stereomicroscopy, with survival and hatching rates recorded. Some blastocysts were stained with 4,6-diamidino2- phenylindole and processed for cytoskeletal evaluation. Three cytoskeletal patterns were identified: Grade I, intact cytoskeleton; Grade II, gross maintenance of integrity, but with some clumps of actin within the cytoplasm; and Grade III, a highly disrupted cytoskeleton. There were no differences in the survival, hatching and cell death rats, total cell number or cytoskeletal integrity between the different vitrification groups. Cell death was greater for vitrified blastocysts than for fresh blastocysts (3.6 +/- 0.4% v. 0.4 +/- 0.7%, respectively; P less than 0.05) and the percentage of blastocysts with a Grade I cytoskeletal pattern was lower for vitrified compared with fresh blastocysts (60.8% v. 92%, respectively; P less than 0.05). The vitrified-warmed blastocysts that hatched during culture exhibited a Grade I cytoskeletal pattern. In conclusion, successful SOPS vitrification of porcine blastocysts does not require pretreatment with cytochalasin B and/or centrifugation.
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| 6. |
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| 7. |
- Guo, Yongzhi, et al.
(författare)
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159 changes in protein expression profiles in bovine endometrial epithelial cells (bEEC) following E coli lipopolysaccharide challenge
- 2014
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 27, s. 170-170
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- E. coli is one of the most frequent bacteria involved in uterine diseases. Lipopolysaccharide (LPS) is a component of the outer membrane of Gram-negative bacteria involved in the pathogenic processes leading to postpartum metritis and endometritis in cattle. It also causes inflammation of the endometrium. Increase of cell proliferation by LPS is part of the inflammatory process and has been reported in human epithelial and immune cells (Martin et al. 2000 J. Immunol. 165, 139-147) and from bovine endometrial epithelial cells (bEEC) (Guo et al. 2014 Reprod. Fertil. Dev. 26, 165-166). The aim of this study was to investigate possible changes in protein expression in relation with the proliferative response of bEEC after challenge with E. coli-LPS. In vitro culture of bEEC was performed from 3 cows. On passage 5, bEEC from each individual were exposed to 0, 8, and 16µgmL-1 LPS for 72h. At time 0 and 72h later, attached cells were counted and for each time and LPS dosage, cells were frozen for proteomic analyses. The variation of cells number over time was analysed by ANOVA (SAS 9.1, proc GLM; SAS Institute, Inc., Cary, NC, USA). All samples were analysed (every sample run in triplicate) by 2-D gel electrophoresis coupled to matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF)/time-of-flight (TOF) mass spectrometry (MS) and shotgun nLC-MS/MS analysis. As reported before, a significant increase in cell number was observed for cells treated with 8µgmL-1 LPS (P≤0.001), whereas changes in cell number were highly variable and nonsignificant for 16µgmL-1 LPS. From each sample, ~800 proteins were visualised. Results from 2-D gel coupled to MALDI-TOF/TOF were very reproducible (same responses between individual cows) and revealed changes in protein profiles very much related (from P<0.05 to P<0.01) to proliferative phenotypes for seven proteins. From shotgun analysis, 27 proteins were found significantly differentially expressed (P<0.05 to P<0.01) following exposure to LPS (21 up-regulated and 6 down-regulated). Among the 21 found as up-regulated, 20 were differentially expressed both for the 8 and 16µgmL-1 LPS, whereas 5 out of 6 were down-regulated for both dosages. Differentially expressed proteins were associated to cell proliferation, apoptosis, oxidative stress, regulation of histones, allergy, and general cell metabolism pathways. Candidate proteins need to be confirmed from larger series of individuals and relevant pathways further studied.
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| 8. |
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| 9. |
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| 10. |
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| 11. |
- Humblot, Patrice
(författare)
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An integrated approach to bovine oocyte quality: from phenotype to genes
- 2016
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 28, s. 1276-1287
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Tidskriftsartikel (refereegranskat)abstract
- In cattle, early embryonic failure plays a major role in the limitation of reproductive performance and is influenced by genetic effects. Suboptimal oocyte quality, including an inadequate store of maternal factors, is suspected to contribute to this phenomenon. In the present study, 13 Montbeliarde cows were phenotyped on oocyte quality, based on their ability to produce viable embryos after in vitro maturation, fertilisation and culture for 7 days. This discriminated two groups of animals, exhibiting developmental rates below 18.8% or above 40.9% (relative to cleaved embryos). Using microarrays, transcriptomic profiles were compared between oocytes collected in vivo from these two groups of animals. The difference in oocyte development potential was associated with changes in transcripts from 60 genes in immature oocytes and 135 genes in mature oocytes (following Bonferroni 5% correction). Of these, 16 and 32 genes were located in previously identified fertility quantitative trait loci. A subset of differential genes was investigated on distinct samples by reverse transcription-quantitative polymerase chain reaction. For SLC25A16, PPP1R14C, ROBO1, AMDHD1 and MEAF6 transcripts, differential expression was confirmed between high and low oocyte potential animals. Further sequencing and searches for polymorphisms will pave the way for implementing their use in genomic selection.
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| 12. |
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| 13. |
- Humblot, Patrice
(författare)
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Dietary propylene glycol and in vitro embryo production after ovum pick-up in heifers with different anti-Mullerian hormone profiles
- 2015
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 27, s. 1249-1261
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Tidskriftsartikel (refereegranskat)abstract
- Rapid genetic improvement in cattle requires the production of high numbers of embryos of excellent quality. Increasing circulating insulin and/or glucose concentrations improves ovarian follicular growth, which may improve the response to superovulation. The measurement of anti-Mu " llerian hormone (AMH) can help predict an animal's response to superovulation treatment. The aim of the present study was to investigate whether increasing circulating insulin concentrations, through propylene glycol (PG) drenches, could improve in vitro embryo production in oestrussynchronised superovulated heifers with different AMH profiles. Holstein heifers were grouped according to preexperimental AMH concentrations as low (L) or high (H). The PG drench increased circulating insulin and glucose concentrations and reduced beta-hydroxybutyrate and urea concentrations compared with the control group. AMH was a good predictor of follicle and oocyte numbers at ovum pick-up (OPU), and of oocyte and embryo quality (AMHH > AMHL). PG in theAMHHgroup increased the number of follicles and blastocyst quality above that in the control group, but did not improve these parameters in theAMHL group. These results indicate that short-term oral PG supplementation modifies an animal's metabolic milieu and is effective in improving in vitro embryo production, after superovulation-OPU, more markedly in heifers with high rather than low AMH concentrations.
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| 14. |
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| 15. |
- Humblot, Patrice
(författare)
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Oral propylene glycol modifies follicular fluid and gene expression profiles in cumulus-oocyte complexes and embryos in feed-restricted heifers
- 2018
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 30, s. 417-429
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Tidskriftsartikel (refereegranskat)abstract
- Dietary supplementation with propylene glycol (PG) increases in vitro production of high-quality embryos in feed-restricted heifers. The aim of the present study was to evaluate the effects of PG in feed-restricted heifers on follicular fluid insulin and insulin-like growth factor (IGF) 1 concentrations, expression of IGF system genes in oocytes and cumulus cells and the expression of selected genes in blastocysts. Feed-restricted (R) heifers were drenched with water or PG during induced oestrous cycles (400 mL of PG or water/drench, daily drenching at 1600 hours for the first 9 days of the oestrous cycle). Ovum pick-up (OPU) was performed after superovulation to produce in vitro embryos and without superovulation to recover oocytes, cumulus cells and follicular fluid. OPU was also performed in a control group (not feed restricted and no drenching). Follicular fluid IGF1 concentrations were reduced by R, and PG restored IGF1 concentrations to those seen in the control group. In cumulus cells, expression of IGF1, IGF1 receptor (IGF1R) and IGF binding protein 4 (IGFBP4) was decreased in the R group, and fully (IGF1 and IGF1R) or partially (IGFBP4) restored to control levels by PG. Blastocyst perilipin 2 (PLIN2; also known as adipophilin), Bcl-2-associated X protein (BAX), SCL2A1 (facilitated glucose/fructose transporter GLUT1), aquaporin 3 (AQP3), DNA (cytosine-5)-methyltransferase 3A (DNMT3A) and heat shock 70-kDa protein 9 (HSPA9B) expression were decreased in R heifers; PG restored the expression of the last four genes to control levels. In conclusion, these results suggest that, during follicular growth, PG exerts epigenetic regulatory effects on gene expression in blastocyst stage embryos.
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| 16. |
- Kruse, Robert, et al.
(författare)
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Colloid centrifugation removes seminal plasma and cholesterol from boar spermatozoa
- 2011
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Ingår i: Reproduction, Fertility and Development. - : CSIRO Publishing. - 1031-3613 .- 1448-5990. ; 23, s. 858-865
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Tidskriftsartikel (refereegranskat)abstract
- The objective of the present study was to investigate the effect of Single-Layer Centrifugation (SLC) on boar spermatozoa, namely the effect of removal of seminal plasma proteins and cholesterol from the surface of spermatozoa. The presence of porcine seminal plasma proteins I and II (PSP-I/PSP-II) before and after SLC was studied using immunofluorescence, whereas the removal of cholesterol was shown qualitatively by thin-layer chromatography (TLC). Finally, the integrity of the sperm plasma membrane was observed by electron microscopy. It was shown that the seminal plasma proteins PSP-I and -II were removed from spermatozoa during SLC but could be restored by adding seminal plasma to the SLC-selected sperm samples. Some cholesterol was also lost from the spermatozoa during SLC but the plasma membrane itself appeared to be morphologically intact. Further studies are underway to examine the relevance of these findings to boar sperm cryopreservation and sperm fertility.
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| 17. |
- Kumaresan, A., et al.
(författare)
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Quantification of kinetic changes in protein tyrosine phosphorylation and cytosolic Ca2+ concentration in boar spermatozoa during cryopreservation
- 2012
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 24:4, s. 531-542
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Tidskriftsartikel (refereegranskat)abstract
- Protein tyrosine phosphorylation in sperm is associated with capacitation in several mammalian species. Although tyrosine phosphorylated proteins have been demonstrated in cryopreserved sperm, indicating capacitation-like changes during cryopreservation, these changes have not yet been quantified objectively. We monitored tyrosine phosphorylation, intracellular calcium and sperm kinematics throughout the cryopreservation process, and studied the relationships among them in boar spermatozoa. Sperm kinetics changed significantly during cryopreservation: curvilinear velocity, average path velocity and straight line velocity all decreased significantly (P < 0.05). While the percentage of sperm with high intracellular calcium declined (P < 0.05), global phosphorylation increased significantly (P < 0.01). Specifically, cooling to 5 degrees C induced phosphorylation in the spermatozoa. After cooling, a 32-kDa protein not observed in fresh semen appeared and was consistently present throughout the cryopreservation process. While the level of expression of this phosphoprotein decreased after addition of the second extender, frozen-thawed spermatozoa showed an increased expression. The proportion of sperm cells with phosphorylation in the acrosomal area also increased significantly (P < 0.05) during cryopreservation, indicating that phosphorylation might be associated with capacitation-like changes. These results provide the first quantitative evidence of dynamic changes in the subpopulation of boar spermatozoa undergoing tyrosine phosphorylation during cryopreservation.
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| 18. |
- Kumaresan, Arumugam, et al.
(författare)
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Sperm function during incubation with oestrus oviductal fluid differs in bulls with different fertility
- 2017
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 29, s. 1096-1106
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Tidskriftsartikel (refereegranskat)abstract
- Spermatozoa undergo several modifications in the oviduct before acquiring fertilising capacity. Although spermatozoa are exposed to similar conditions in the oviduct, the speed of the response varies with the male and the state of the spermatozoa. We hypothesised that spermatozoa from bulls with different fertility may differ in their ability to respond to oviductal fluid (ODF). Frozen-thawed spermatozoa from four bulls were incubated with oestrus oviductal fluid (OODF) for 6h. Sperm kinematics, tyrosine phosphorylation, phosphorylation patterns, capacitation and acrosome reaction were analysed at hourly intervals. The amplitude of lateral head displacement (ALH) and straightness coefficient (STR) were higher (P < 0.05) in bulls with higher fertility compared with those with lower fertility, at 1-4h of incubation. At 4h of incubation and onwards, spermatozoa from bulls with higher fertility showed a lower degree (P < 0.05) of tyrosine phosphorylation and higher degree of capacitation and acrosome reaction. At least five tyrosine-phosphorylated sperm proteins were detected in all bulls. However, the expression of two phosphorylated sperm proteins (183 and 109 kDa) was upregulated in bulls with lower fertility. It may be concluded that cryopreserved spermatozoa from high- and low-fertile bulls differ in their ability to respond to OODF. This may help in developing tools for assessing fertility of bulls, once validated in more animals.
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| 19. |
- Laskowski, Denise, et al.
(författare)
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Insulin exposure during in vitro bovine oocyte maturation changes blastocyst gene expression and developmental potential
- 2017
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 29, s. 876-889
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Tidskriftsartikel (refereegranskat)abstract
- Metabolic imbalance impairs fertility, because changes in concentrations of metabolites and hormones in the blood and follicular fluid create an unfavourable environment for early embryonic development. Insulin is a key metabolic hormone known for its effects on fertility: insulin concentrations are increased during energy balance disturbances in diabetes or metabolic syndrome. Still, insulin is frequently used at supraphysiological concentrations for embryo in vitro culture with unknown consequences for the developmental potential of the offspring. In the present study we investigated the effects of insulin exposure during in vitro bovine oocyte maturation on developmental rates, embryo quality and gene expression. Supplementation of the maturation media with insulin at 10 or 0.1 mu gmL(-1) decreased blastocyst rates compared with an insulin-free control (19.8 +/- 1.3% and 20.4 +/- 1.3% vs 23.8 +/- 1.3%, respectively; P<0.05) and led to increased cell numbers (nearly 10% more cells on Day 8 compared with control; P<0.05). Transcriptome analysis revealed significant upregulation of genes involved in lipid metabolism, nuclear factor (erythroid-derived 2)-like 2 (NRF2) stress response and cell differentiation, validated by quantitative polymerase chain reaction. To conclude, the results of the present study demonstrate that insulin exposure during in vitro oocyte maturation has a lasting effect on the embryo until the blastocyst stage, with a potential negative effect in the form of specific gene expression perturbations.
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| 20. |
- Laskowski, Denise, et al.
(författare)
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Insulin treatment during in vitro oocyte maturation leads to different gene expression and methylation patterns of key genes associated with metabolism and steroid synthesis in the bovine blastocyst
- 2017
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 29, s. 108-
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Obesity and overfeeding are common causes for female infertility, leading to insulin resistance and hyperinsulinemia and associated with an increased risk for type 2 diabetes mellitus (Pasquali et al., http://dx.doi.org/10.1093/humupd/dmg024). We investigated here the effect of insulin during in vitro oocyte maturation on methylation changes in bovine Day 8 blastocysts (BC8) and focused on methylation patterns of candidate genes associated with metabolism and steroidogenesis (Day 0¼day of oocyte collection). Abattoir-derived oocytes (n¼882) were in vitro matured for 22 h with 2 different insulin concentrations, INS10 (10 mgmL 1) and INS0.1 (0.1 mgmL 1) or without insulin (INS0, control). Subsequently, IVF and IVC were performed to equal standardized conditions for all groups. Parallel genomic DNA and total RNA extraction (AllPrepDNA/RNA micro kit, cat no. 80284, Qiagen, Valencia, CA, USA) from pools of 10 frozen (808C) BC8 was followed by transcriptome and epigenome analysis (Laskowski et al., http://dx.doi.org/10.1071/RD15315).An empirical Bayesmoderated t-test and the ‘limma' package in R (www.r-project.org) were used to search for differentially expressed genes between the control and the insulin groups.Analysis of the epigenome by using a specific pipeline, described by Shojaei Saadi et al. (2014 BMC Genomics 15, 451), showed that 7632 and 3914 regionswere hypomethylated in the INS0.1 and INS10 v. INS0, whereas 6026 and 8504 regions were hypermethylated in INS0.1 and INS10 v. INS0. Combining epigenetic and transcriptomic data,we found that highmethylation and lowexpression or the reverse (lowmethylation and high expression) were observed for a set of 14 and 11 genes for INS0.1 and INS10 respectively. Most of these genes are associated with lipid metabolism, steroid synthesis, and oxidative stress. Further investigation of the localization of differentially methylated regions (DMR) in genes showed that the conservation odds (methylation) was in general higher in coding regions and CpG islands than in noncoding regions. We observed a large overlap of DMR in the 2 insulin groups compared with controls (3233 common DMR). These numerous changes illustrate the potential unfavourable effects of elevated insulin during maturation leading to alteration of the methylation patterns of the early embryo. This model may help us better understand the mechanisms by which metabolic disorders observed pre-conception can affect embryonic development and subsequent health of the offspring. Our results based on changes in transcriptome or epigenome did showthat insulin challenge during maturation leads to postponed effects associated with steroidogenesis, lipid metabolism and oxidative stress in the BC8. By this early stage, if persistent, specific changes in the expression and methylation patterns of genes associated to hyperinsulinemia may decrease the developmental potential of early embryos or could be responsible for subsequent pathologies.
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| 21. |
- Laskowski, Denise, et al.
(författare)
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Lipid profile of bovine blastocysts exposed to insulin during in vitro oocyte maturation
- 2018
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 30, s. 1253-1266
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Tidskriftsartikel (refereegranskat)abstract
- Insulin is a key hormone with important functions in energy metabolism and is involved in the regulation of reproduction. Hyperinsulinaemia is known to impair fertility (for example, in obese mothers); therefore, we aimed to investigate the impact of elevated insulin concentrations during the sensitive period of oocyte maturation on gene expression and lipid profiles of the bovine Day-8 embryo. Two different insulin concentrations were used during in vitro oocyte maturation (INS10 = 10 mu g mL(-1) and INS0.1 = 0.1 mu g mL(-1)) in order to observe possible dose-dependent effects or thresholds for hyperinsulinaemia in vitro. By investigating gene expression patterns by an mRNA microarray in combination with lipid profile analysis by desorption electrospray ionisation-mass spectrometry (DESI-MS) of embryos derived from insulin-treated oocytes, we gained further insights regarding molecular responses of embryos to insulin provocation during the first days of development. Lipid metabolism appeared to be influenced on multiple levels according to gene expression results but the profiles collected in positive-ion mode by DESI-MS (showing mostly ubiquinone, cholesteryl esters and triacylglycerols) did not differ significantly from controls. There are parallels in follicular development of ruminants and humans that make this bovine model relevant for comparative research on early human embryonic development during hyperinsulinaemia.
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| 22. |
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| 23. |
- Martinez-Madrid, Belen, et al.
(författare)
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Immunolocalisation of aquaporins 3, 7, 9 and 10 in the epididymis of three wild ruminant species (Iberian ibex, mouflon and chamois) and sperm cryoresistance
- 2023
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Ingår i: Reproduction, Fertility and Development. - : CSIRO PUBLISHING. - 1031-3613 .- 1448-5990. ; 35:16, s. 708-721
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Tidskriftsartikel (refereegranskat)abstract
- Context In the epididymis, epithelial cells manage changes in the luminal environment for proper sperm maturation. Moreover, aquaglyceroporins, a subgroup of aquaporins (AQP), modulate the transport of water, glycerol and other small molecules in epithelial cells.AimsWe aim to characterise the lining epithelium, quantify its cell composition and immunolocalise the aquaglyceroporins AQP3, AQP7, AQP9 and AQP10 alongside the epididymal ductus of three wild ruminant species, and to determine if species-specific differences could be associated with cauda sperm cryoresistance variations.MethodsEpididymides from Iberian ibex (n = 5), mouflon (n = 5) and chamois (n = 6) were obtained. Cauda spermatozoa were collected and sperm parameters were analysed before and after freezing. Histology and immunohistochemistry of AQP3, 7, 9, 10 and T-CD3 were performed in the caput, corpus and cauda epididymal regions.Key resultsThis work first describes the lining epithelium in Iberian ibex, mouflon and chamois epididymis along the three anatomical regions, consisting of principal, basal, apical, clear and halo cells. However, the percentage of each cell type differed in ibex compared to mouflon and chamois. The positive T-CD3 immunolabeling of all the halo cells confirmed their T-lymphocyte nature. Aquaglyceroporin expression patterns were similar among species, except for differences in AQP7 and AQP10 immunolocalisation in ibex. Species-specific differences in epididymal sperm cryoresistance were confirmed.ConclusionsThe epididymal epithelium of the three wild ruminants differ in their relative number of cell types and AQP immunolocalisation, which ultimately appears to affect cauda epidydimal spermatozoa cryoresistance.ImplicationsOur study provides information on the relevance of the quantitative composition and AQP pattern expression in epididymal lining epithelium on sperm cryoresistance. This work describes the lining epithelium and expression patterns of aquaglyceroporins (AQP3, AQP7, AQP9 and AQP10) in the Iberian ibex, mouflon and chamois epididymis. Species-specific differences in cauda epididymal sperm cryoresistance were confirmed. Differences among the three wild ruminant epididymides in the relative number of epithelial cell types and the immunolocalisation of AQP7 and AQP10 appear to affect the cryoresistance of cauda epididymal spermatozoa.
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| 24. |
- Mattsson, Anna, et al.
(författare)
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Effects on differentiation of reproductive organs and sexual behaviour in Japanese quail by excessive embryonic ERα activation
- 2010
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 22:2, s. 416-425
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Tidskriftsartikel (refereegranskat)abstract
- Exposure of Japanese quail (Coturnix japonica) embryos to oestrogenic substances disrupts sexual differentiation of the reproductive tract of both sexes and impairs the copulatory behaviour of the adult male. To examine whether these effects can be induced by selective activation of oestrogen receptor alpha (ER alpha), Japanese quail eggs were injected with various doses of the selective ER alpha agonist 16 alpha-lactone-oestradiol (16 alpha-LE2). The natural oestrogen 17 beta-oestradiol (E-2) was used as a positive control. Both 16 alpha-LE2 and E-2 induced formation of an ovary-like cortex in the left testis (ovotestis) and reduced the size of the right testis in male embryos. The asymmetry in testis size remained in sexually mature males. Both substances induced retention and malformation of the Mullerian ducts in embryos of both sexes and malformed oviducts in juveniles. Male copulatory behaviour was suppressed by embryonic exposure to E-2 and the highest dose of 16 alpha-LE2. However, the lower dose of 16 alpha-LE2, which markedly affected development of the reproductive organs, was without effects on behaviour. It can therefore not be excluded that the behavioural demasculinisation at the 100-fold higher dose involved cross-activation of oestrogen receptor beta (ER beta). In conclusion, our results suggest that oestrogen-induced disruption of reproductive organ development in Japanese quail can be mediated via ER alpha, whereas the role of ER alpha in demasculinisation of copulatory behaviour remains to be clarified.
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| 25. |
- Morrell, Jane
(författare)
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Differences in preservation of canine chilled semen using simple sperm washing, single-layer centrifugation and modified swim-up preparation techniques
- 2016
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 28, s. 1545-1552
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Tidskriftsartikel (refereegranskat)abstract
- This study compared the efficacy of simple sperm washing (SW), single-layer centrifugation (SLC) and modified swim-up (SU) techniques in the preparation of dog spermatozoa for cooling. Eighteen ejaculates, collected from three dogs (six per dog), were pooled (three ejaculates per pool) and divided into three aliquots: (1) one aliquot was washed and cooled at 5 degrees C for 72h, considered as control (SW-control), (2) the second aliquot was selected by SLC through Androcoll-C and subsequently cooled in the same way as the SW-control samples (SLC-AC) and (3) the last aliquot was selected by a modified SU method with Androcoll-C and cooled as mentioned above (SU-AC). Assessment of sperm motility, sperm morphology, sperm membrane integrity and acrosome integrity were performed on aliquots of fresh semen and chilled-rewarmed samples. Sperm membrane integrity and progressive motility were significantly (P<0.05) improved by SU-AC compared with SW-control. Morphological sperm abnormalities decreased significantly (P<0.001) in SLC-AC samples compared with SW-control samples. These sperm variables did not differ between SLC-AC and SU-AC methods (P>0.05). The recovery rates were not significantly (P>0.05) different between SW-control, SLC-AC and SU-AC samples. Our results confirm that SU-AC may be a successful method for the preparation of dog spermatozoa for cooling.
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| 26. |
- Morrell, Jane
(författare)
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Donor sperm production in heterologous recipients by testis germ cell transplantation in the dromedary camel
- 2019
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 31, s. 538-546
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Tidskriftsartikel (refereegranskat)abstract
- The object of this study was to investigate if testis germ cell transplantation (TGCT) into a heterologous recipient would result in donor-origin spermatogenesis in the dromedary camel. First, we investigated a workable protocol for TGCT in camels, including donor cell isolation, enrichment by density gradient centrifugation (Percoll and Bovicoll), rete testis injection and microsatellite detection of donor and recipient genotypes. Second, the effects of three doses of Dolichos biflorus agglutinin (DBA), a glycoprotein that specifically binds to gonocytes or Type A spermatogonia, on testis germ cell depletion were investigated by direct injection into the rete testis of a male camel. Seven recipients were prepared with DBA treatment, two males were castrated at 4 weeks for depletion assessment and the remaining five received donor cells 4-6 weeks after treatment. On average, similar to 17 million cells were isolated per gram of testis tissue, with 19.5 +/- 1.9% DBA-positive (DBA(+)) cells. Percoll centrifugation yielded a 1.5-fold increase in DBA(+) cells while Bovicoll centrifugation produced a 2.5-fold increase from the input cells of 18.6 +/- 2.1% DBA(+) cells. Semen was collected from the recipients 13-20 weeks after transfer and the presence of donor DNA in the samples was determined using microsatellite markers. In two of the five recipients, all semen samples were shown to be positive for donor-derived cells. These results demonstrate for the first time that: (1) heterologous testicular germ cell transplantation in camels is feasible and the recipients are able to produce spermatozoa of donor origin and (2) DBA can be used effectively to deplete endogenous stem cells.
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| 27. |
- Morrell, Jane, et al.
(författare)
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Effect of Single Layer Centrifugation on reactive oxygen species and sperm mitochondrial membrane potential in cooled stallion semen
- 2017
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 29, s. 1039-1045
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Tidskriftsartikel (refereegranskat)abstract
- Additional means are needed for evaluating the quality of stallion spermatozoa in semen doses for AI. Mitochondrial membrane potential ((m)) has been linked to fertility in some species, but is rarely used in the evaluation of cooled stallion semen; metabolic activity may be associated with reactive oxygen species production (ROS). In the present study, (m) and ROS production were measured in doses of cooled stallion semen. The effect of colloid centrifugation on these parameters was also investigated. In this case, colloid centrifugation involves centrifuging a sperm sample through a silane-coated silica colloid formulation to retrieve the most robust spermatozoa. High and low (m) in cooled stallion semen varied between stallions and between ejaculates, but was not affected by single-layer centrifugation (SLC). The SLC-selected spermatozoa produced significantly less hydrogen peroxide than controls (P<0.001), which could explain the increased longevity and retention of fertilising capacity seen in previous studies. For SLC samples, (m) was positively associated with viable spermatozoa that were not producing reactive oxygen species (r=0.49; P<0.001) and negatively associated with ROS production (for superoxide: r=-0.4, P<0.01; for hydrogen peroxide: r=-0.39, P<0.05). There was no clear association between (m) and ROS production in control samples.
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| 28. |
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| 29. |
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| 30. |
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| 31. |
- Norrby, Mattias
(författare)
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Restricted feed intake in lactating primiparous sows. II. Effects on subsequent litter sex ratio and embryonic gene expression
- 2011
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Ingår i: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 23, s. 899-911
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Tidskriftsartikel (refereegranskat)abstract
- Expression of panels of candidate genes controlling myogenesis, angiogenesis and gender-specific imprinting of development were analysed in embryonic, placental and endometrial tissues recovered at Day 30 of gestation from a subset of primiparous sows that were either feed restricted (Restrict; n=17) or fed to appetite (Control; n=15) during the last week of the previous lactation. Embryos were also sex typed to investigate gender bias in response to treatments. Average embryonic weight was lower in the subset of Restrict compared with Control litters (1.38 +/- 0.07 vs 1.59 +/- 0.08 g, respectively) and the male : female sex ratio was higher (P<0.05) in embryos (litters) recovered from Restrict sows. Treatment affected (P <= 0.05) the expression of embryonic and placental genes involved in insulin-like growth factor (IGF) 2 signalling, including IGF2, INSR and IGF2R. Embryonic expression of ESR1 was also affected by treatment (P<0.03) and sex x treatment interactions were observed for the expression of embryonic ESR1 (P<0.05) and placental ANGPT2 (P<0.03). At the molecular level, these results support the suggestion that changes in placental function are not the primary mechanism mediating detrimental effects of previous sow catabolism on early embryonic development in the feed-restricted lactational sow model. However, perturbations in the IGF2 system are implicated as mediators of these effects.
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| 32. |
- Rodriguez-Martinez, Heriberto
(författare)
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State of the art in farm animal sperm evaluation
- 2007
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Ingår i: Reproduction, Fertility and Development. - : CSIRO Publishing. - 1031-3613 .- 1448-5990. ; 19:1, s. 91-101
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Tidskriftsartikel (refereegranskat)abstract
- Our ability to screen the structural and functional integrity of the spermatozoon in vitro has increased markedly over the past decades, but our capacity to estimate the fertility of a semen sample or of the sire from which it has been collected, especially in selected farm animal breeders, has not. The estimation of fertility is constrained by several factors (e.g. type of cell, analysis strength, sperm deposition strategies, recordings of fertility), including the fact that the ejaculate is composed of a diverse sperm population. Such cell heterogeneity is reflected not only in differences in the intactness of attributes needed for fertilisation, such as motility or morphology, but also in the relative ability of the spermatozoa to remain fertile over time, to sustain selection steps and responses to exogenous stimuli similar to those during sperm transport in the female genital tract, all of which account for innate variations in the fertilising ability among doses, ejaculates and sires. Determination of how large such a sperm population with competence for fertilisation and in-built ability to display these attributes under physiological signalling is would allow for a better estimation of fertility, provided that the particular sire produces this sub-population in a repeatable manner. The value of these analyses is discussed in the present paper.
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| 33. |
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| 34. |
- Tsiartas, P., et al.
(författare)
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Seven days ex vivo perfusion of whole ewe ovaries with follicular maturation and oocyte retrieval: towards the development of an alternative fertility preservation method
- 2022
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Ingår i: Reproduction Fertility and Development. - : CSIRO Publishing. - 1031-3613 .- 1448-5990. ; 34:3, s. 331-342
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Tidskriftsartikel (refereegranskat)abstract
- Fertility preservation methods for prepubertal women about to undergo gonadotoxic chemo and/or radiation therapy are limited. Therefore, the aim of this study was to investigate the feasibility to develop an alternative fertility preservation method based on an ex vivo perfusion platform for whole ewe ovaries. Thirteen ewe ovaries were divided into two groups (group 1 and 2) that were perfused in a bioreactor for up to 7 days. Group 1 (n = 3) were stimulated with human menopausal gonadotropin (hMG) administered in single daily dose, while group 2 (n = 10) were stimulated continuously for 24 h. The perfused ovaries in group 1 showed no significant differences in follicular density, sub-follicular morphology and oocyte quality after ischaemia and after ex vivo perfusion compared with non-perfused control ovaries. The perfused ovaries in group 2 showed a significant decrease in the follicular reserve and oocyte quality compared with the control group. In total, 16 GV-MI oocytes were retrieved from both groups. This study describes for the first time the ex vivo maintenance of viable follicles of ewe ovaries with oocyte integrity and the retrieval of oocytes after ex vivo hormonal perfusion with two different protocols for up to 7 days.
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| 35. |
- Bage, R, et al.
(författare)
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Deviant peri-oestrual hormone patterns affect the epithelium of the uterine tube in repeat-breeder heifers
- 2002
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Ingår i: Reproduction, fertility, and development. - : CSIRO Publishing. - 1031-3613. ; 14:87-8, s. 461-469
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Tidskriftsartikel (refereegranskat)abstract
- In the bovine reproductive tract, the uterine tube is the critical site for a series of events required for fertilization and early embryonic development. In previous studies, a defined category of subfertile heifers, repeat-breeder heifers (RBH), has presented peri-oestrual disturbances (deviating hormone patterns and follicular dynamics) and uterine maternal–embryonic asynchrony. The present study aimed to investigate if tubal function was also affected, by determination of differences in the morphology of the tubal lining epithelium of RBH (n = 4) in comparison to controls (n = 6) during standing oestrus, studied by light and electron microscopy (SEM/TEM), and relate this to steroid hormone concentrations and receptor distribution in the target tissues. Tissue distribution of oestrogen receptor α (ERα) and progesterone receptor B (PRB) was quantified using immunohistochemistry. In particular, secretory cells differed in appearance between RBH and controls. The cells were less lumen protruding, microvilli were fewer and smaller and secretory granules in the apical cytoplasm were more numerous in RBH. Furthermore, the tubal epithelium was conspicuously coated with amorphous material. Morphological differences between categories were not explained hormonally or by steroid receptor distribution, except in two heifers from which uterine tubes were obtained after the luteinizing hormone (LH) surge. The isthmic PRB : ERα ratio was twice as high in the RBH than in the control. The deviating ultrastructure found in RBH, before and after the LH surge, might influence the tubal microenvironment with effects on gamete transport and final maturation and early embryonic development. The present study confirms that previously recorded perturbations in reproductive physiology in RBH are also manifested in the uterine tube, mainly by a deviating ultrastructure of the lining epithelium.
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| 36. |
- Familari, Mary, et al.
(författare)
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Placenta-derived extracellular vesicles : Their cargo and possible functions
- 2017
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Ingår i: Reproduction, Fertility and Development. - 1031-3613. ; 29:3, s. 433-447
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Forskningsöversikt (refereegranskat)abstract
- The literature on extracellular vesicles consists of rapidly expanding and often contradictory information. In this paper we attempt to review what is currently known regarding extracellular vesicles released specifically from human placental syncytiotrophoblast cells with a focus on the common but complex pregnancy-associated syndrome pre-eclampsia, where the level of syncytiotrophoblast extracellular vesicle release is significantly increased. We review common methods for syncytiotrophoblast extracellular vesicle derivation and isolation and we discuss the cargo of syncytiotrophoblast extracellular vesicles including proteins, RNA and lipids and their possible functions. A meta-analysis of available trophoblast-derived extracellular vesicle proteomic datasets revealed only three proteins in common: albumin, fibronectin-1 and plasminogen activator inhibitor-1, suggesting some variability in vesicle cargo, most likely reflecting stage and cell type of origin. We discuss the possible sources of variability that may have led to the low number of common markers, which has led us to speculate that markers and density in common use may not be strict criteria for identifying and isolating placenta-derived exosomes.
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| 37. |
- Fernández-Gago, Rocío, et al.
(författare)
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Thawing boar semen in the presence of seminal plasma improves motility, modifies subpopulation patterns and reduces chromatin alterations.
- 2017
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Ingår i: Reproduction, fertility, and development. - : CSIRO PUBLISHING. - 1031-3613. ; 29:8, s. 1576-1584
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Tidskriftsartikel (refereegranskat)abstract
- Seminal plasma could have positive effects on boar semen after thawing. In the present study we investigated changes in the motility and chromatin structure in spermatozoa over 4h incubation (37°C) of boar semen thawed in the presence of 0%, 10% or 50% seminal plasma from good-fertility boars. Cryopreserved doses were used from seven males, three of which were identified as susceptible to post-thawing chromatin alterations. Motility was analysed by computer-aided sperm analysis every hour, and data were used in a two-step clustering, yielding three subpopulations of spermatozoa (slow non-linear, fast non-linear, fast linear). Chromatin structure was analysed using a sperm chromatin structure assay and flow cytometry to determine the DNA fragmentation index (%DFI) as a percentage, the standard deviation of the DFI (SD-DFI) and the percentage of high DNA stainability (%HDS), indicating chromatin compaction. Thawing without seminal plasma resulted in a rapid loss of motility, whereas seminal plasma helped maintain motility throughout the incubation period and preserved the subpopulation comprising fast and linear spermatozoa. The incidence of chromatin alterations was very low in samples from non-susceptible males, whereas samples from males susceptible to post-thawing chromatin alterations exhibited marked alterations in%DFI and%HDS. Seminal plasma partly prevented these alterations in samples from susceptible males. Overall, 50% seminal plasma was the most efficient concentration to protect motility and chromatin. Some changes were concomitant with physiological events reported previously (e.g., semen thawed with 50% seminal plasma increased the production of reactive oxygen species and yielded higher fertility after AI). Thawing in the presence of seminal plasma could be particularly useful in the case of samples susceptible to post-thawing chromatin damage.
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| 38. |
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| 39. |
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| 40. |
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| 41. |
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| 42. |
- Kolakowska, J., et al.
(författare)
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Proteomic analysis of the endometrium during early pregnancy in the domestic pig
- 2017
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Ingår i: Reproduction Fertility and Development. - : CSIRO Publishing. - 1031-3613. ; 29:11, s. 2255-2268
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Tidskriftsartikel (refereegranskat)abstract
- Reproductive processes in domestic pigs have been studied extensively. Pigs are one of the main sources of meat for human consumption and are an established model for investigations into mammalian, including human, reproductive physiology. Studies of the uterus during early pregnancy will lead to a better understanding of mechanisms governing pregnancy. Proteomics provides the possibility to explore endometrial functions in an unbiased way. The aim of the study was to compare endometrium harvested from Days 12-13 and 15-16 of pregnancy with the corresponding days of the oestrous cycle. We identified endometrial proteins that are unique to the early stages of pregnancy (Days 12-13 and 15-16). Twenty-one proteins were identified that were uniquely expressed on the selected days of pregnancy or the oestrous cycle. Out of 21 identified proteins, 14 referred to the pregnancy periods. Systemic analysis of the identified proteins revealed cell adhesion and cytoskeletal organisation as two of the major functions, both of which are important for the establishment and maintenance of pregnancy. Thrombospondin 1 expression was validated using western blotting analysis and the results suggest its involvement in the adhesiveness of the embryo during the peri-implantation period in pigs.
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| 43. |
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| 44. |
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| 45. |
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| 46. |
- Tienthai, Paisan, et al.
(författare)
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Expression of hyaluronan synthase-3 in porcine oviducal epithelium during oestrus.
- 2003
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Ingår i: Reprod Fertil Dev. - 1031-3613. ; 15:1-2, s. 99-105
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Tidskriftsartikel (refereegranskat)abstract
- Hyaluronan (HA) has been related to fertilization and embryo development in the pig. Furthermore, HA is present in pig oviduct fluid and the lining epithelium, particularly of the pre-ovulatory sperm reservoir. Because the mechanisms that regulate HA synthesis have not yet been clarified, semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was conducted to assess the expression of mRNAs of two HA-synthesizing enzymes (has2 and has3) in the oviduct epithelium (uterotubal junction, isthmus, ampullary-isthmic junction and ampulla segments) of non-inseminated (control) and inseminated (treatment) sows at pre-, peri- and post-ovulatory oestrus. Only has3 mRNA was detected; it was present in all tubal segments of both control and treatment samples. The level of has3 expression did not vary significantly between non-inseminated and inseminated specimens, but there was a tendency (NS) for increased mean values during the peri- and post-ovulatory stages compared with pre-ovulation. It is concluded that has3 is expressed by the porcine endosalpinx epithelium and the levels of expression do not vary during the critical periods of sperm transport and fertilization, despite fluctuating levels of HA in the tubal fluid at corresponding periods.
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| 47. |
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| 48. |
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