| 1. |
- Atikuzzaman, Mohammad, 1977-
(författare)
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Seminal Influence on the Oviduct : Mating and/or semen components induce gene expression changes in the pre-ovulatory functional sperm reservoir in poultry and pigs
- 2016
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Internal fertilization occurs in birds and eutherian mammals. Foetal development, however, is either extra- respectively intra-corpore (egg vs uterus). In these animal classes, the female genital tract stores ejaculated spermatozoa into a restricted oviductal segment; the functional pre-ovulatory sperm reservoir, where they survive until ovulation/s occur. Paradoxically, this immunologically foreign sperm suspension in seminal fluid/plasma, often microbiologically contaminated, ought to be promptly eliminated by the female local immune defence which, instead, tolerates its presence. The female immune tolerance is presumably signalled via a biochemical interplay of spermatozoa, as well as the peptides and proteins of the extracellular seminal fluid, with female epithelial and immune cells. Such interplay can result in gene expression shifts in the sperm reservoir in relation to variations in fertility. To further aid our understanding of the underlying mechanisms, this thesis studied the proteome of the seminal fluid (using 2D SDS-PAGE and mass spectrometry) including cytokine content (using Luminex and/or ELISA) of healthy, sexually mature and fertile boars and cocks. As well, gene expression changes (using cDNA microarray) in the oviductal sperm reservoirs of sexually-mature females, mated or artificially infused with homologous sperm-free seminal fluid/plasma were studied. Pigs were of commercial, fertility-selected modern breeds (Landrace), while chicken belonged to the ancestor Red Junglefowl (RJF, low egg laying-capacity), a selected egg-layer White Leghorn (WL) and of their Advanced Intercross Line (AIL). Ejaculates were manually collected as single sample in cocks or as the sperm-rich fraction [SRF] and the post- SRF fraction in boars to harvest seminal fluid/plasma for proteome/cytokine and infusion-studies. Oviducts were retrieved for gene-expression analyses via microarray immediately post-mortem (chicken) or at surgery (pig), 24 h after mating or genital infusion. In pigs, the protein-rich seminal plasma showed the highest amounts of cytokines [interferon-γ, interferon gamma-induced protein 10 (IP-10/CXCL10), macrophage derived chemokine (MDC/CCL22), growth-regulated oncogene (GRO/CXCL1), granulocyte-macrophage colony-stimulating factor (GM-CSF), monocyte chemo-attractant protein-1 (MCP-1/ CCL2), interleukin (IL)-6, IL-8/CXCL8, IL-10, IL-15, IL-17 and transforming growth factor (TGF)-β1-3) in the larger, protein-rich and sperm-poor post-SRF, indicating its main immune signalling influence. Chicken showed also a plethora of seminal fluid proteins with serum albumin and ovotransferrin being conserved through selection/evolution. However, they showed fewer cytokines than pigs, as the anti-inflammatory/immune-modulatory TGF-β2 or the pro-inflammatory CXCL10. The RJF contained fewer immune system process proteins and lacked TGF-β2 compared to WL and AIL, suggesting selection for increased fertility could be associated with higher expression of immune-regulating peptides/proteins. The oviductal sperm reservoir reacted in vivo to semen exposure. In chicken, mating significantly changed the expression of immune-modulatory and pH-regulatory genes in AIL. Moreover, modern fertile pigs (Landrace) and chicken (WL), albeit being taxonomically distant, shared gene functions for preservation of viable sperm in the oviduct. Mating or SP/SF-infusion were able to change the expression of comparable genes involved in pH-regulation (SLC16A2, SLC4A9, SLC13A1, SLC35F1, ATP8B3, ATP13A3) or immune-modulation (IFIT5, IFI16, MMP27, ADAMTS3, MMP3, MMP12). The results of the thesis demonstrate that both mating and components of the sperm-free seminal fluid/plasma elicit gene expression changes in the pre-ovulatory female sperm reservoir of chickens and pigs, some conserved over domestication and fertility-selection.
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| 2. |
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| 3. |
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| 4. |
- Broström, Hans, et al.
(författare)
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Topical treatment of equine sarcoids with imiquimod 5% cream or Sanguinaria canadensis and zinc chloride - an open prospective study
- 2020
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Ingår i: Veterinary Dermatology. - : Wiley. - 0959-4493 .- 1365-3164. ; 31, s. 471-481
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Tidskriftsartikel (refereegranskat)abstract
- Background Equine sarcoids are the most prevalent skin neoplasm in horses worldwide. Although several treatments are available, none are consistently effective and recurrence is common. Objectives To evaluate the efficacy and safety of topical imiquimod 5% cream andSanguinaria canadensis + zinc chloride for treatment of equine sarcoids and investigate possible systemic effects on distant untreated sarcoids. Animals/Tumours Twenty-five client-owned horses with a total of 164 tumours were included in the study. Fifty-seven tumours were treated and 107 tumours were left untreated. Methods and materials Biopsies were taken from a minimum of one tumour per horse and the rest were diagnosed based on clinical appearance as likely sarcoids. Imiquimod 5% (A) was applied three times weekly, whileSanguinaria canadensis + zinc chloride (X) was applied every fourth day after a six day daily initiation phase. Treatment continued until clinical remission or for a maximum of 45 weeks, with a long follow-up period (mean 34 months). Biopsies of sarcoids were re-taken before treatment termination and at follow-up if the owner gave consent. Results Complete remission was recorded in 84.4% (A) and 75.0% (X) of the tumours. Relapse was recorded in 7.3% (A) and 21.4% (X). Spontaneous remission was observed in 1.9% of untreated tumours. No systemic effect on untreated tumours was detected. During treatment varying degrees of local inflammatory reaction were common. Conclusions and clinical relevance Both treatments were considered effective and safe. Smaller tumours responded more favourably to treatment. Relapse rate was low and not observed in sarcoids with repeat biopsies before treatment termination.
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| 7. |
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| 8. |
- Chankeaw, Wiruntita, et al.
(författare)
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Analysis of the transcriptome of bovine endometrial cells isolated by laser micro-dissection (1): specific signatures of stromal, glandular and luminal epithelial cells
- 2021
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Ingår i: BMC Genomics. - : Springer Science and Business Media LLC. - 1471-2164. ; 22
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Tidskriftsartikel (refereegranskat)abstract
- Background A number of studies have examined mRNA expression profiles of bovine endometrium at estrus and around the peri-implantation period of pregnancy. However, to date, these studies have been performed on the whole endometrium which is a complex tissue. Consequently, the knowledge of cell-specific gene expression, when analysis performed with whole endometrium, is still weak and obviously limits the relevance of the results of gene expression studies. Thus, the aim of this study was to characterize specific transcriptome of the three main cell-types of the bovine endometrium at day-15 of the estrus cycle. Results In the RNA-Seq analysis, the number of expressed genes detected over 10 transcripts per million was 6622, 7814 and 8242 for LE, GE and ST respectively. ST expressed exclusively 1236 genes while only 551 transcripts were specific to the GE and 330 specific to LE. For ST, over-represented biological processes included many regulation processes and response to stimulus, cell communication and cell adhesion, extracellular matrix organization as well as developmental process. For GE, cilium organization, cilium movement, protein localization to cilium and microtubule-based process were the only four main biological processes enriched. For LE, over-represented biological processes were enzyme linked receptor protein signaling pathway, cell-substrate adhesion and circulatory system process. Conclusion The data show that each endometrial cell-type has a distinct molecular signature and provide a significantly improved overview on the biological process supported by specific cell-types. The most interesting result is that stromal cells express more genes than the two epithelial types and are associated with a greater number of pathways and ontology terms.
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| 9. |
- Chankeaw, Wiruntita, et al.
(författare)
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Analysis of the transcriptome of bovine endometrial cells isolated by laser micro-dissection (2): impacts of post-partum negative energy balance on stromal, glandular and luminal epithelial cells
- 2021
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Ingår i: BMC Genomics. - : Springer Science and Business Media LLC. - 1471-2164. ; 22
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Tidskriftsartikel (refereegranskat)abstract
- Background In post-partum dairy cows, the energy needs to satisfy high milk production induces a status of more or less pronounced Negative Energy Balance (NEB). NEB associated with fat mobilization impairs reproductive function. In a companion paper, we described constitutive gene expression in the three main endometrial cell types (stromal, glandular and luminal epithelial cells) isolated by laser capture micro-dissection (LCM) showing the specificities of their transcriptomic profiles. This study investigates the specific impact of NEB on gene expression in these cells around 80 days after parturition at day 15 of the oestrus cycle and describes their specific response to NEB. Results Following the description of their constitutive expression, the transcriptome profiles obtained by RNA sequencing of the three cells types revealed that differences related to the severity of NEB altered mainly specific patterns of expression related to individual cell types. Number of differentially expressed genes between severe NEB (SNEB) and mild NEB (MNEB) cows was higher in ST than in LE and GE, respectively. SNEB was associated with differential expression of genes coding for proteins involved in metabolic processes and embryo-maternal interactions in ST. Under-expression of genes encoding proteins with functions related to cell structure was found in GE whereas genes encoding proteins participating in pro-inflammatory pathways were over-expressed. Genes associated to adaptive immunity were under-expressed in LE. Conclusion The severity of NEB after calving is associated with changes in gene expression around 80 days after parturition corresponding to the time of breeding. Specific alterations in GEs are associated with activation of pro-inflammatory mechanisms. Concomitantly, changes in the expression of genes encoding proteins involved in cell interactions and maternal recognition of pregnancy takes place in ST. The combination of these effects possibly altering the uterine environment and embryo maternal interactions may negatively influence the establishment of pregnancy.
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| 10. |
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| 11. |
- 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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| 12. |
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| 13. |
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| 14. |
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| 15. |
- Chanrot, Metasu, et al.
(författare)
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Bovine herpes virus type 4 alters TNF-α and IL-8 profiles and impairs the survival of bovine endometrial epithelial cells
- 2017
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Ingår i: Reproductive Biology. - : Elsevier BV. - 1642-431X .- 2300-732X. ; 17, s. 225-232
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Tidskriftsartikel (refereegranskat)abstract
- Bovine herpes virus type 4 (BoNV-4) can be transmitted by contaminated semen to cows at the time of breeding and may cause uterine disease. The aim of this study was to characterize the susceptibility of bovine endometrial epithelial cells (bEEC) to BoHV-4 by using an in vitro model. When bEEC were challenged with different multiplicity of infection (MOI; from 0.001 to 10) of BoHV-4 for 6 days, a significant decrease in cell survival with increasing MOI was observed. The bEEC were subsequently challenged with BoNV-4 MOI 0.1 for 7 days. During the first 4 days, numbers increased in a similar way in controls and infected group (p < 0.01 when compared to Day 0). After Day 4, numbers of live cells in infected samples decreased when compared to controls and were lower than control at Day 7 (p < 0.01). From titration and qPCR, increasing number of viral particles was observed from Day 1, and reached a plateau at Day 5. Concentrations of IL-8 increased with time and were higher in supernatants from infected cells than in controls (p < 0.0001). TNF-alpha concentrations presented similar profile as cell survival ones. In conclusion, the survival of bEEC was strongly impaired by BoNV-4 infection in a time and dose dependent manner and supernatant cytokine profiles were altered. This information supports BoHV-4 implication in clinical cases of uterine diseases and the existence of a risk of BoNV-4 transmission from infected males through animal breeding. (C) 2017 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier Sp. z o.o. All rights reserved.
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| 16. |
- Chanrot, Metasu, et al.
(författare)
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Dose related effects of LPS on endometrial epithelial cell populations from dioestrus cows
- 2017
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Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 177, s. 12-24
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Tidskriftsartikel (refereegranskat)abstract
- Lipopolysaccharides (LPS) from Gram negative bacteria are involved in the pathogeny of uterine diseases in cows. This study aimed to investigate LPS effects on the growth of bovine endometrial epithelial cells (bEEC) and relationships between LPS response and tissue characteristics. Uteri from 35 females were characterized for parity and stage of oestrous cycle. Densities of glandular tissue (dGT), CD11b+ cells and Ki67+ cells were measured in the endometrial tissue. Cells from 13 dioestrus cows were exposed to 0, 2, 4, 8, 12, 16 or 24 mu g/mL LPS. Effects of parity and stage of the oestrous cycle on tissue characteristics and effects of LPS dosage, cow and tissue characteristics on changes in cell numbers were analyzed by ANOVA. The dGT was higher in metoestrus and dioestrus samples than in pro-oestrus ones whereas densities of CD11b+ and Ki67+ cells were higher at pro-oestrus (p < 0.05-p < 0.01). LPS influenced bEEC populations in a dose related manner. An increase in number of live cells was observed for dosages ranging from 2 to 12 mu g/mL LPS (p < 0.0001 vs controls). No effect was found on numbers and frequencies of dead cells. With higher dosages, the numbers of live cells did not increase but the numbers of dead did increase. No relationships were observed between cow or tissue characteristics and growth patterns or frequencies of viable bEEC in controls nor in the response to LPS. To conclude this model is suitable for further studies on dysregulations induced by LPS in endometrial tissue. (C) 2016 The Authors. Published by Elsevier B.V.
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| 17. |
- 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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| 18. |
- Guo, Yongzhi, et al.
(författare)
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Adipokines as biomarkers of postpartum subclinical endometritis in dairy cows
- 2020
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Ingår i: Reproduction. - 1470-1626 .- 1741-7899. ; 160, s. 417-430
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Tidskriftsartikel (refereegranskat)abstract
- Adipokines emerged as regulators of metabolism and inflammation in several scenarios. This study evaluated the relationship between adipokines (adiponectin, chemerin and visfatin) and cytological (subclinical) endometritis, by comparing healthy (without), transient (recovered by 45 days postpartum (DPP)) and persistent (until 45 DPP) endometritis cows (n = 49). Cows with persistent endometritis had higher adiponectin concentrations in plasma (at 21 DPP, P < 0.05 and at 45 DPP, P < 0.01) and in uterine fluid (at 45 DPP, P < 0.001), and higher chemerin concentrations in plasma (P < 0.05) and uterine fluid (P < 0.01) at 45 DPP than healthy cows. Cows with persistent endometritis had higher gene transcription in the cellular pellet of uterine fluid and protein expression in the endometrium of these adipokines and their receptors than healthy cows. Adiponedin plasma concentrations allowed to discriminate healthy from persistent endometritis cows, in 87% (21 DPP) and 98% (45 DPP) of cases, and adiponectin and chemerin uterine fluid concentrations at 45 DPP allowed for this discrimination in 100% of cases. Cows with concentrations above the cutoff were a minimum of 3.5 (plasma 21 DPP), 20.4 (plasma 45 DPP), and 33.3 (uterine fluid 45 DPP) times more at risk of evidencing persistent endometritis at 45 DPP than cows with concentrations below the cutoff. Overall, results indicate a relationship between adipokine signalling and the inflammatory status of the postpartum uterus of dairy cows, evidencing that adipokines represent suitable biomarkers of subclinical endometritis, able to predict the risk of persistence of inflammation.
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| 19. |
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| 20. |
- Guo, Yongzhi, et al.
(författare)
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Changes in protein expression profiles in bovine endometrial epithelial cells exposed to E. coli LPS challenge
- 2017
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Ingår i: Molecular BioSystems. - : Royal Society of Chemistry (RSC). - 1742-206X .- 1742-2051. ; 13, s. 392-405
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Tidskriftsartikel (refereegranskat)abstract
- E. coli is one of the most frequently involved bacteria in uterine diseases. Lipopolysaccharide (LPS) is a component of the outer membrane of Gram-negative bacteria involved in pathogenic processes leading to post-partum metritis and endometritis in cattle. It also causes inflammation of the endometrium. The increase of cell proliferation by LPS is part of the inflammatory process. The aim of this study was to investigate possible changes in protein expression in relation to the proliferative response of bEECs after challenge with E. coli-LPS. In vitro culture of bEECs was performed from cow genital tracts collected at a slaughterhouse. In passage 5, bEECs from each of 9 cows (3 series of 3 cows) were exposed to 0, 8, and 16 mg ml(-1) LPS for 72 h. At time 0 and 72 h later, attached cells/living cells were counted and for each time and LPS dosage, cells were frozen for proteomic analyses. All samples from the 3 series were analyzed by 2-D gel electrophoresis coupled to MALDI-TOF/TOF mass spectrometry. The samples from the first series were subjected to shotgun nLC-MS/MS analysis. From the whole differential proteomics analysis, 38 proteins were differentially expressed (p < 0.05 to p < 0.001) following exposure to LPS. Among them, twenty-eight were found to be up-regulated in the LPS groups in comparison to control groups and ten were down-regulated. Differentially expressed proteins were associated with cell proliferation and apoptosis, transcription, destabilization of cell structure, oxidative stress, regulation of histones, allergy and general cell metabolism pathways. The de-regulations induced by LPS were consistent with the proliferative phenotype and indicated strong alterations of several cell functions. In addition, some of the differentially expressed proteins relates to pathways activated at the time of implantation. The specific changes induced through those signals may have negative consequences for the establishment of pregnancy.
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| 21. |
- Guo, Yongzhi, et al.
(författare)
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Characterization of circulating microRNA profiles of postpartum dairy cows with persistent subclinical endometritis
- 2023
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Ingår i: Journal of Dairy Science. - 0022-0302 .- 1525-3198. ; 106, s. 9704-9717
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Tidskriftsartikel (refereegranskat)abstract
- Subclinical endometritis (SCE) is an unresolved in-flammation of the endometrium of postpartum dairy cows, seriously affecting fertility. Current diagnosis, which relies on uterine cytology or even more invasive biopsy sampling, would benefit from the identifica-tion of blood-based diagnostic biomarkers. Due to the known role of microRNAs (miRNAs) in other diseases, this case-control study evaluated the cell-free circulat-ing miRNA profiles of SCE cows, and the network transcripts predicted to interact with those miRNAs, previously identified as differentially expressed genes (DEG) in the endometrium of the same cows. Healthy (H, n = 6) and persistent SCE (n = 11) cows character-ized by endometrial cytology and biopsy were blood sampled at 21 and 44 d postpartum (DPP). Following extraction of cell-free plasma miRNAs and RNA-seq analysis, differential abundance analysis of miRNAs was performed with the DESeq2 R package (adjusted p-value of 0.05), and in silico prediction of miRNA-interacting genes on a sequence complementary basis was conducted using the miRWalk database. The principal component analysis showed a clear cluster-ing between groups of uterine health phenotypes (H vs. SCE), although the clustering between groups was less pronounced at 44 DPP than at 21 DPP. No effect the stage (21 vs. 44 DPP) was observed. A total of 799 known circulating miRNAs were identified, from which 34 demonstrated differential abundance between H and SCE cows (12 less abundant and 22 more abundant SCE than in H cows). These 34 miRNAs are predicted to interact with 10,104 transcripts, among which 43, 81, and 147 were previously identified as differentially expressed in, respectively, endometrial luminal epithe- lial, glandular epithelial, and stromal cells of the same cows. This accounts for approximately half of the DEG identified between those H and SCE cows, including genes involved in endometrial cell proliferation, angio- genesis and immune response, whose dysregulation in SCE cows may impair pregnancy establishment. From 219 miRNAs with mean normalized read counts above 100, the presence and abundance of miR-425-3p and miR-2285z had the highest discriminatory level to dif- ferentiate SCE from H cows. In conclusion, despite ap- parent confinement to the endometrium, SCE is associ- ated with a distinct circulating miRNA profile, which may represent a link between the systemic changes associated with disease and the endometrial immune response. The validation of a miRNA panel consist- ing of circulating cell-free miR-425-3p and miR-2285z may prove a relevant advancement for the noninvasive diagnosis of persistent SCE.
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| 22. |
- Guo, Yongzhi, et al.
(författare)
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Differential gene expression in bovine endometrial epithelial cells after challenge with LPS; specific implications for genes involved in embryo maternal interactions
- 2019
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Ingår i: PLoS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 14
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Tidskriftsartikel (refereegranskat)abstract
- Lipopolysaccharide (LPS) expressed on the surface of Gram-negative bacteria activates pro-inflammatory pathways, dys-regulates the function of endometrial cells and is a key player in the mechanisms involved in endometritis. This study aimed to investigate the effects of LPS on bovine endometrial epithelial cells (bEEC) from whole transcriptome with a special focus on genes involved in embryo-maternal interactions. Following in vitro culture, bEEC from three cows were exposed to 0, 2, and 8 mu g/mL LPS for 24h. RNA samples extracted at 0 and 24 hours were analyzed by RNA sequencing (RNA-seq). At 24h, 2035 differentially expressed genes (DEGs) were identified between controls and samples treated with 2 mu g/mL LPS. Gene ontology analysis showed that over-expressed DEGs were associated to immune response, response to stress and external stimuli, catalytic activity, and cell cycle. Genes associated with cell membrane and cell adhesion pathways were under-expressed. LPS induced changes in expression of specific genes related to embryo-maternal interactions including under-expression of eight members of the cadherin superfamily, over-expression of six members of the mucin family, and differential expression of a large set of genes binding the above molecules and of more than 20 transcripts coding for cyto-kines and their receptors. Type I interferon-tau dependent genes were also over-expressed. From a sub-set of 19 genes, (biological replicates of bEEC from cows taken at time 6 (n = 3), 24 (n = 6) and 48 hours (n = 3), and 2 technical replicates per sample) differential gene expression was confirmed by RT2-qPCR (r(2) between fold changes at 24 hours by RT2 qPCR and RNA-seq = 0.97). These results indicate that LPS affects the function of bEEC in many ways by differential transcription, glycolytic metabolism and oxidative stress. Many transcriptomic signatures related to implantation and embryo maternal interactions were strongly affected by LPS. These results pave the way for further studies to investigate the duration of these changes and their possible impact on endometrial function and fertility.
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| 23. |
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| 24. |
- Guo, Yongzhi, et al.
(författare)
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Progesterone differentially affects the transcriptomic profiles of cow endometrial cell types
- 2022
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Ingår i: BMC Genomics. - : Springer Science and Business Media LLC. - 1471-2164. ; 23
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Tidskriftsartikel (refereegranskat)abstract
- Background The endometrium is a heterogeneous tissue composed of luminal epithelial (LE), glandular epithelial (GE), and stromal cells (ST), experiencing progesterone regulated dynamic changes during the estrous cycle. In the cow, this regulation at the transcriptomic level was only evaluated in the whole tissue. This study describes specific gene expression in the three types of cells isolated from endometrial biopsies following laser capture microdissection and the transcriptome changes induced by progesterone in GE and ST cells. Results Endometrial LE, GE, and ST cells show specific transcriptomic profiles. Most of the differentially expressed genes (DEGs) in response to progesterone are cell type-specific (96%). Genes involved in cell cycle and nuclear division are under-expressed in the presence of progesterone in GE, highlighting the anti-proliferative action of progesterone in epithelial cells. Elevated progesterone concentrations are also associated with the under-expression of estrogen receptor 1 (ESR1) in GE and oxytocin receptor (OXTR) in GE and ST cells. In ST cells, transcription factors such as SOX17 and FOXA2, known to regulate uterine epithelial-stromal cross-talk conveying to endometrial receptivity, are over-expressed under progesterone influence. Conclusions The results from this study show that progesterone regulates endometrial function in a cell type-specific way, which is independent of the expression of its main receptor PGR. These novel insights into uterine physiology present the cell compartment as the physiological unit rather than the whole tissue.
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| 25. |
- Guo, Yongzhi, et al.
(författare)
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Subclinical endometritis differentially affects the transcriptomic profiles of endometrial glandular, luminal, and stromal cells of postpartum dairy cows
- 2022
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Ingår i: Journal of Dairy Science. - : American Dairy Science Association. - 0022-0302 .- 1525-3198. ; 105, s. 6125-6143
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Tidskriftsartikel (refereegranskat)abstract
- In postpartum dairy cows, subclinical endometritis flammation, which has profound detrimental effects on subsequent reproductive performance. To date, transcriptomic studies related to this condition were either based on biopsy-derived whole-endometrium tissue or endometrial swab or cytobrush samples, thus masking effects of disease on cell type-specific gene expression. This study tested the hypothesis that different endometrial health statuses are associated with distinct transcription profiles of endometrial stromal, glandular, and luminal epithelial cells. At 44 d postpartum (DPP), endometrial biopsies were taken from dairy or affected by persistent SCE, according to endometrial cytology taken at 21 and 44 DPP. Stromal, glandular, and luminal epithelial cells were isolated from the whole-tissue biopsy by laser capture microdissection, and the cell-specific transcription profiles were determined by RNA sequencing. Differential gene expression demonstrated that global transcriptomic profiles and corresponding lists of differentially expressed genes beamong cell types. Results also showed that although healthy and recovered cows presented similar endoprior presence of immune cells still affected the transcriptome of endometrial cells at this stage, delaying complete functional recovery. Recovery or persistence of inflammation was associated with gene expression patterns involved not only in immune function but also in tissue remodeling, cell adhesion, and uterine recep-tivity in a cell type-specific manner. Identifying these signatures may contribute to the development of novel diagnostic tools and therapeutic strategies. In addition, these results may help to define preventive measures or ways to stimulate recovery from endometrial inflamma-tion, thus helping to restore the fertility of postpartum dairy cows.
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| 26. |
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| 27. |
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| 28. |
- 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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| 29. |
- Humblot, Patrice
(författare)
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Antral follicle count, oocyte production and embryonic developmental competence of senescent Nellore (Bos indicus) cows
- 2021
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 174, s. 27-35
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Tidskriftsartikel (refereegranskat)abstract
- Information on the follicular population and oocyte quality of cows in the final period of reproductive life is scarce. The present study aimed to compare the antral follicle count (AFC), oocyte production and embryonic developmental competence of young versus long-lived and senescent Bos indicus beef cows. Nellore cows (Bos indicus) were classified into three groups according to age: young (4-9 years, n = 10), long-lived (14-17 years, n = 10) and senescent (17-23 years, n = 10). At a random time in the estrus cycle, the cows received cloprostenol sodium salt (0.5 mg, IM), estradiol benzoate (1 mg, IM) and an intravaginal P4 device (1.4 g). Five days later, the P4 devise was removed and oocyte collection (OPU1) was performed. A second OPU (OPU2) was performed 5 days after the first in order to aspirate only growing follicles. During each OPU, AFC and the number and quality of cumulus-oocyte complexes (COCs) were evaluated. Then, the COCs were placed in standard maturation medium (IVM), fertilized and incubated for 9 days. The data were subjected to ANOVA and Multinomial Logistic Regression. The AFC was smaller in long-lived and senescent cows in both OPU1 and OPU2 when compared to younger cows. There was no difference in AFC between OPU1 (19.9 +/- 1.8) and OPU2 (17.6 +/- 1.9) in young cows, however, more follicles were punctured in long-lived and senescent cows in OPU1 (12.0 +/- 2.6 and 19.3 +/- 4.6) than in OPU2 (9.2 +/- 1.9 and 10.3 +/- 2.3), respectively (P < 0.01). The numbers of COCs recovered from young cows (OPU1 = 14.2 +/- 1.8; OPU2 = 8.4 +/- 0.9) were higher than those obtained from long-lived cows (OPU1 = 5.9 +/- 2.3; OPU2 = 4.3 +/- 1.0) and senescent cows (OPU1 = 7.2 +/- 3.0; OPU2 = 4.1 +/- 1.7), respectively (P < 0.05). The cleavage rate did not differ between groups. However, the rate of blastocyst formation was higher for young (64.8%) and long-lived (65.0%) compared to senescent (16.5%) cows (P 0.01). In conclusion our results indicate that the AFC is lower in long-lived and senescent cows compared with young cows. However, unlike in senescent cows, the embryonic development of long-lived cows is similar to that of young cows. This suggests that Nellore cows aged 17 years begin to have reduced embryonic development capacity due to ovarian aging. (c) 2021 Elsevier Inc. All rights reserved.
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| 30. |
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| 31. |
- 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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| 32. |
- Humblot, Patrice
(författare)
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Differences of Periods of Calving on Days Open of Dairy Cows in Different Regions and Months of Thailand
- 2011
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Ingår i: Thai Journal Of Veterinary Medicine. - 0125-6491. ; 41, s. 315-320
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Tidskriftsartikel (refereegranskat)abstract
- The objectives of this study were to analyze the potential impact of heat stress on reproduction in different regions of Thailand, to determine the monthly distribution of calving throughout the year and to investigate environmental sources of variation of days-open (DO) in first lactation dairy cows. Data including 13,548 lactation records were collected during 2004 to 2006. Climate data were obtained from the provincial meteorological stations and the corresponding temperature-humidity indexes were calculated. The difference between regions in THI was determined. The geographical regions studied were: Central, Eastern, Northeastern, and Northern. The distribution of calving by month was determined in the 1st to 5th lactation. The effect of month of calving (MOC) on DO was determined only in first lactation dairy cows. The fixed effects in the model included MOC, region and MOC x region. The lowest mean THI was observed in December (72) and the highest mean THI in April (80). THI differed significantly between regions (p<0.0001), and months (p<0.0001). Significant interactions between region and month (p<0.0001) were found for THI. In all regions, minimum THI values were observed in December and January and this effect was more pronounced in the Northeastern and Northern regions. The highest frequency of calving for the first lactation was observed in June (9.96%) and the lowest in February (6.63%). The highest frequencies of calving for the 2(nd) (13.1%), 3(rd) (14.1%), and 4(th) (14.66%) lactation cows were observed in September while for the 5th lactation cows was in October (14.91%). The lowest proportion of calving for 2(nd) (5.02%) and 4(th) (4.14%) lactation cows was in February, and in March for the 3(rd) (4.35%) and 5(th) (4.85%) lactation cows. The average DO in first lactation cows was 152 days. Significant effects of MOC (p<0.0001.) and region (p<0.0001.) were found on DO. February calving cows had the longest DO (219 +/- 11 days) while cows calving in October and November had a significantly shorter mean DO (133 +/- 7 days). The study indicates that the high proportion of cows calving in October and November corresponds to breeding success in the previous months of December and January, which are the coolest Months of the year. Cows which calved during hot months had a prolonged DO period of several months.
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| 33. |
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| 34. |
- Humblot, Patrice
(författare)
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Effects of a dietary supplement of B-carotene given during the dry period on milk production and circulating hormones and metabolites in dairy cows
- 2012
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Ingår i: Revue de Médecine Vétérinaire. - 0035-1555. ; 163, s. 235-241
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Tidskriftsartikel (övrigt vetenskapligt/konstnärligt)abstract
- The objective of this study was to investigate whether a supplement of beta-carotene given during the dry period is able to 1) improve milk production and milk composition and 2) modify hormone and metabolic status in dairy cows during the dry and postpartum periods. This study was conducted using 40 Holstein, primiparous and multiparous cows. On the day of drying-off, cows were allocated to one of two dietary treatments: control diet (n= 20) or control diet plus 1g/cow/d beta-carotene (n= 20). The beta-carotene supplement was given individually to the cows until calving. Blood samples were obtained regularly and the concentrations of beta-carotene in blood and circulating metabolites and hormones in plasma were measured. Live weight and body condition score (BCS) were monitored once a month. Daily milk production was recorded starting 5 days after calving. Milk composition was measured every 15 days. The dietary supplement of beta-carotene increased blood concentrations of beta-carotene during the dry period (P< 0.05) and although the difference decreased postpartum beta-carotene concentrations remained higher compared to the control group until the end of experiment. Live weight, BCS, milk production and composition: milk protein, milk fat, milk urea and somatic cell count, were unaffected by treatment (P> 0.05). Plasma concentrations of insulin, insulin-like growth factor-1, glucose, non-esterified fatty acids and urea were unaffected by dietary supplementation with beta-carotene (P> 0.05). In conclusion, supplementation with beta-carotene during the dry period increased blood concentrations of beta-carotene but had no effect on milk production and milk quality nor on hormone and metabolic status.
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| 35. |
- Humblot, Patrice
(författare)
-
Effects of embryo size at transfer (whole versus demi) and early pregnancy progesterone supplementation on embryo growth and pregnancy-specific protein bovine concentrations in recipient dairy heifers
- 2011
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 76, s. 522-531
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Tidskriftsartikel (refereegranskat)abstract
- The objectives of this study were to evaluate embryonic size and survival, plasma progesterone (P4) and pregnancy-specific protein bovine (PSPB) concentrations in early pregnancies (n = 99) following the transfer of one whole (n = 66) or one demi (n = 33) embryo to recipient virgin dairy heifers. The experiment was designed to evaluate the fixed effects of embryo size at transfer (whole or demi embryo) on Day 7 of the estrous cycle (Day 0 = estrus) and P4 supplementation between Days 7 to 19 through an intravaginal device (yes or no) on plasma P4 and PSPB concentrations and on embryo measurements. Plasma P4 concentrations were measured by RIA on Days 0, 7, 14, 19, 21, 25, 35, 42, 49, 56 and 63 of pregnancy and, PSPB concentrations were measured by ELISA on Days 7, 21, 25, 35, 42, 49, 56 and 63. The presence of an embryonic vesicle was detected on Day 25, embryonic/fetal movements and heartbeat were evaluated on Days 42 and 63 and embryo measurements [crown-rump length (CRL) and width at mid body] were obtained on Day 42 through ultrasonography.In non-supplemented pregnancies, Day 42 whole embryos had higher (P < 0.05) CRL and width than demi embryos, but the difference averaged only 1 to 2 mm. In P4 supplemented pregnancies, whole and demi embryos attained a similar size on Day 42 of pregnancy. Embryo size at transfer, early exogenous P4 supplementation and their interactions had no effects (P > 0.05) on plasma P4 concentrations. However, the post-hoc LSD evaluation showed that plasma P4 concentrations on Day 25 were higher (P < 0.001) in whole than in demi embryo derived pregnancies and, that exogenous P4 supplementation increased (P < 0.05) plasma P4 concentrations on Day 19 of pregnancy. The plasma PSPB detection rate on Days 7 to 63 of pregnancy was similar in pregnancies resulting from the transfer of whole and demi embryos. From a total of 93 recipients remaining pregnant until Day 63, plasma PSPB was constantly undetectable on Day 7, was detected in 4% of Day 21 samples, 41% of Day 25, 95% of Day 35, 96% of Day 42, 99% of Day 49 and in 100% of samples of Days 56 and 63. Concentrations of PSPB increased (P < 0.05) from Days 21 to 42 and from Days 56 to 63, with a plateau between Days 42 to 56. Demi embryo pregnancies had higher (P < 0.05) plasma PSPB concentrations on Days 35 and 42 than whole embryo pregnancies. Progesterone supplementation had a positive effect (P < 0.01) on PSPB concentrations from Days 35 to 63. Concentrations of PSPB were similar in non-supplemented whole and demi embryo pregnancies from Days 7 to Day 63. In contrast, in supplemented recipients, demi embryo pregnancies had higher (P < 0.05) PSPB concentrations on Days 25 to 42 than whole embryo pregnancies. No significant correlation was found between P4 and PSPB concentrations or between the concentrations of these hormones and embryonic measurements on Day 42. In conclusion, demi embryos experienced a compensatory growth until Day 42 of pregnancy, attaining a similar size to that of whole embryos and originating conceptuses producing similar plasma PSPB concentrations to those of whole embryo derived conceptuses. Embryonic growth and conceptus secretion of PSPB were positively stimulated by early pregnancy exogenous P4 treatment. (C) 2011 Elsevier Inc. All rights reserved.
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| 36. |
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| 37. |
- Humblot, Patrice, et al.
(författare)
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Factors affecting the accuracy of pregnancy scanning in ewes
- 2013
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Ingår i: Veterinary Record. - : Wiley. - 0042-4900 .- 2042-7670. ; 173, s. 606-
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Tidskriftsartikel (refereegranskat)abstract
- The objective was to evaluate the accuracy of pregnancy scanning by transabdominal ultrasonography and affecting factors. Altogether 44,783 ewes were registered (2008-2010), 39,724 diagnosed as pregnant and subsequently lambed. The ewes, 0.5-14 years old, were divided into 4 age and 6 breed groups and analysed. The accuracy (per cent; number of scanned fetuses/number of born lambs) decreased with increasing number of fetuses (P<0.001). Overall accuracy was 90.3 per cent, highest (93.7 per cent) in ewes carrying one fetus, 91.9 per cent, 82.4 per cent, and 74.9 per cent in ewes with 2, 3 and >= 4 fetuses at scanning, respectively. Finnsheep ewes with highest number of lambs showed lowest accuracy (P<0.001). Analyses of a more complete sub set of data (n=23,396), showed that number of fetuses diagnosed, breed, age of ewe, operator and time in gestation, significantly affected the accuracy. Accuracy decreased with age of ewe (P<0.001) and was 71.8 per cent, 91.6 per cent and 89.3 per cent for scanning at <40, 40-80 and 81-100 days of gestation, respectively (P<0.001). In general, the numbers of fetuses were overestimated at scanning and increased gradually with number of fetuses diagnosed. In conclusion, the accuracy was affected by several factors, which should be considered when interpreting/implementing the results, especially in breeds with high fecundity.
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| 38. |
- Humblot, Patrice
(författare)
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From clinics to (cow)mics: a reproductive journey
- 2018
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Ingår i: Animal Reproduction. - 1806-9614 .- 1984-3143. ; 15, s. 278-291
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Tidskriftsartikel (refereegranskat)abstract
- This manuscript describes the different topics I have been involved in the fields of reproductive physiology and embryo biotechnologies with attempts to address practical issues raised mainly by the breeding industry. The journey started with phenotyping work in the field of reproductive physio-pathology. Other issues were related to the optimization of reproductive biotechnologies to favorize genetic selection. The implementation of genomic selection raised opportunities to develop the use embryo biotechnologies and showed the interest of combining them in the case of embryo genotyping. There is still a need to refine phenotyping for reproductive traits especially for the identification of markers of uterine dysfunction. It is believed that new knowledge generated by combining different molecular approaches will be the source of applications that may benefit AI practice and embryo technologies.
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| 39. |
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| 40. |
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| 41. |
- Humblot, Patrice
(författare)
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Invited review: Learning from the future-A vision for dairy farms and cows in 2067
- 2018
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Ingår i: Journal of Dairy Science. - : American Dairy Science Association. - 0022-0302 .- 1525-3198. ; 101, s. 3722-3741
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Forskningsöversikt (refereegranskat)abstract
- The world's population will reach 10.4 billion in 2067 with 81% residing in Africa or Asia. Arable land available for food production will decrease to 0.15 ha per person. Temperature will increase in tropical and temperate zones, especially in the Northern Hemisphere, and this will push growing seasons and dairy farming away from arid areas and into more northern latitudes. Dairy consumption will increase because it provides essential nutrients more efficiently than many other agricultural systems. Dairy farming will become modernized in developing countries and milk production per cow will increase, doubling in countries with advanced dairying systems. Profitability of dairy farms will be the key to their sustainability. Genetic improvements will include emphasis on the coding genome and associated noncoding epigenome of cattle, and on microbiomes of dairy cattle and farmsteads. Farm sizes will increase and there will be greater lateral integration of housing and management of dairy cattle of different ages and production stages. Integrated sensors, robotics, and automation will replace much of the manual labor on farms. Managing the epigenome and microbiome will become part of routine herd management. Innovations in dairy facilities will improve the health of cows and permit expression of natural behaviors. Herds will be viewed as superorganisms, and studies of herds as observational units will lead to improvements in productivity, health, and well-being of dairy cattle, and improve the agroecology and sustainability of dairy farms. Dairy farmers in 2067 will meet the world's needs for essential nutrients by adopting technologies and practices that provide improved cow health and longevity, profitable dairy farms, and sustainable agriculture.
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| 42. |
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| 43. |
- Humblot, Patrice
(författare)
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Metabolomic Prediction of Pregnancy Viability in Superovulated Cattle Embryos and Recipients with Fourier Transform Infrared Spectroscopy
- 2014
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Ingår i: BioMed Research International. - : Hindawi Limited. - 2314-6133 .- 2314-6141.
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Tidskriftsartikel (refereegranskat)abstract
- We analyzed embryo culture medium (CM) and recipient blood plasma using Fourier transform infrared spectroscopy (FTIR) metabolomics to identify spectral models predictive of pregnancy outcome. Embryos collected on Day 6 from superovulated cows in 2 countries were individually cultured in synthetic oviduct fluid medium with BSA for 24 h before embryo transfer. Spent CM, blank controls, and plasma samples (Day 0 and Day 7) were evaluated using FTIR. The spectra obtained were analyzed. The discrimination capability of the classifiers was assessed for accuracy, sensitivity (pregnancy), specificity (nonpregnancy), and area under the ROC curve (AUC). Endpoints considered were Day 60 pregnancy and birth. High AUC was obtained for Day 60 pregnancy in CM within individual laboratories (France AUC = 0.751 +/- 0.039, Spain AUC = 0.718 +/- 0.024), while cumulative data decreased the AUC (AUC = 0.604 +/- 0.029). Predictions for CM at birth were lower than Day 60 pregnancy. Predictions with plasma at birth improved cumulative over individual results (Day 0: France AUC = 0.690 +/- 0.044; Spain AUC < 0.55; cumulative AUC = 0.747 +/- 0.032). Plasma generally predicted pregnancy and birth better than CM. These first results show that FTIR metabolomics could allow the identification of embryos and recipients with improved pregnancy viability, which may contribute to increasing the efficiency of selection schemes based on ET.
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| 44. |
- Humblot, Patrice
(författare)
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Oocyte and embryo production and quality after OPU-IVF in dairy heifers given diets varying in their n-6/n-3 fatty acid ratio
- 2012
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 78, s. 632-645
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Tidskriftsartikel (refereegranskat)abstract
- Dietary fat supplementation can improve oocyte quality in ruminants. The influence of the type of dietary fat on the number and quality of oocytes collected by ovum pick-up and on the production of embryos in vitro was investigated in Holstein heifers. Heifers were given hay plus one of two dietary supplements for 42 days. The supplements were linseed (L, rich in linolenic acid, C18:3n-3, n = 9) or soya bean (S, rich in linoleic acid, C18:2n-6, n = 9). Oocytes were collected by ovum pick-up (OPU) for 6 wks (2 sessions/wk) and morphologic quality assessed. Half the oocytes were frozen and the other half was used to produce embryos. Blood samples were analyzed for: insulin, insulin-like growth factor-1, glucose, non-esterified fatty acids, B-hydroxy butyrate and urea and the proportions of fatty acids. Neither growth rate nor plasma hormone and metabolite concentrations were affected by dietary supplement. However, L significantly increased the proportion of plasma C18:3n-3 while S significantly increased the proportion of C18:2n-6(P < 0.001). Neither oocyte characteristics (number, their quality and number fertilized and cleaved per heifer per session) nor embryo characteristics (number and quality per heifer per session) and embryo development stages were affected by dietary treatment. Real-time RT-PCR was performed on immature and mature cumulus-oocyte complexes (COC). Prostaglandin E synthase-1 expression increased in L compared to S heifers. In conclusion, the type of fatty acid did not modify the numbers of oocytes and embryos produced by OPU-IVF and their quality in dairy heifers. Upregulation of prostaglandin E synthase-1 may ensure sufficient POE, production for oocyte maturation even when its precursor is low. (C) 2012 Elsevier Inc. All rights reserved.
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| 45. |
- Humblot, Patrice
(författare)
-
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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| 46. |
- Humblot, Patrice
(författare)
-
Prediction of pregnancy viability in bovine in vitro-produced embryos and recipient plasma with Fourier transform infrared spectroscopy
- 2014
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Ingår i: Journal of Dairy Science. - : American Dairy Science Association. - 0022-0302 .- 1525-3198. ; 97, s. 5497-5507
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Tidskriftsartikel (refereegranskat)abstract
- We analyzed embryo culture medium (CM) and recipient blood plasma using Fourier transform infrared (FTIR) metabolomics to predict pregnancy outcome. Individually cultured, in vitro-produced (IVP) blastocysts were transferred to recipients as fresh and vitrified-warmed. Spent CM and plasma samples were evaluated using FTIR. The discrimination capability of the classifiers was assessed for accuracy, sensitivity (pregnancy), specificity (nonpregnancy), and area under the receiver operator characteristic curve (AUC). Within all IVP fresh embryos (birth rate = 52%), high AUC were obtained at birth, especially with expanded blastocysts (CM: 0.80 +/- 0.053; plasma: 0.89 +/- 0.034). The AUC of vitrified IVP embryos (birth rate = 31%) were 0.607 +/- 0.038 (CM, expanded blastocysts) and 0.672 +/- 0.023 (plasma, all stages). Recipient plasma generally predicted pregnancy outcome better than did embryo CM. Embryos and recipients with improved pregnancy viability were identified, which could increase the economic benefit to the breeding industry.
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| 47. |
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| 48. |
- Humblot, Patrice
(författare)
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Reproductive performance of Bos indicus beef cows treated with different doses of equine chorionic gonadotropin at the end of a progesterone-estrogen based protocol for fixed-time artificial insemination
- 2018
-
Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 118, s. 150-156
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Tidskriftsartikel (refereegranskat)abstract
- Two experiments were performed to evaluate the reproductive performance of zebu beef cows treated with different doses of eCG at the end of a progesterone (P4)/estrogen based protocol for timed artificial insemination (TAI). In Experiment 1, suckling Bos indicus Nelore cows (n = 261) received, on day 0, a progesterone (P4) intravaginal device (PD) and an injection of 1 mg estradiol benzoate (EB). On day 8, the PD was removed, 500 pig of cloprostenol was injected, and cows were assigned to one of the following groups: Control (no treatment), 300 (300 IU of eCG), 600 (600 IU of eCG), and 900 (900 IU of eCG). On day 9, all cows received 1 mg EB and TAI performed 54-56 h after cloprostenol injection. A pregnancy diagnosis was done by ultrasound scanning 40 days after TAI, and the number of fetuses and calves was recorded at pregnancy diagnosis and at birth. More cows treated with eCG displayed estrus within 48 h after removal of the PD (42.3% vs. 11.6%, P < 0.01), and ovulated more than one follicle (42%, 58/138 vs. 1.8%, 1/54; P < 0.01). This effect on ovulation rate was dose dependent (P < 0.05). The pregnancy rate was affected only by cow parity (primiparous, 25.3% vs. multiparous, 48.9%; P < 0.01). Twin pregnancy was higher (P < 0.01) in cows treated with eCG (42%, 58/138) than controls (0%, 0/54). However, few cows (33.3%) were able to keep both fetuses intact until birth. For evaluation of ovarian characteristics by B mode and Doppler ultrasonography, 43 Nelore cows were submitted In Experiment 2 to the same four groups described in Experiment 1. Although no difference (P > 0.1) was observed for size and blood perfusion in the pre-ovulatory follicles, corpus luteum was larger and with greater blood perfusion (P <0.05) in eCG-treated cows. In conclusion, eCG increased the number of double/multiple ovulations in a dose-dependent manner, induced larger and more vascularized corpora lutea, but did not affect the fertility of cyclic or anestrous cows. Although eCG results in twin pregnancies, most of cows underwet embryo/fetus loss and birth a single calf. (C) 2018 Elsevier Inc. All rights reserved.
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| 49. |
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| 50. |
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