Inmaculada ParrillaMaria Antonia GilCristina CuelloJosep M. Cambra...
10页
查看更多>>摘要：The establishment of a successful pregnancy can only occur through a concerted functioning of the entire female reproductive system, allowing for fertilization, subsequent embryo development and implantation of the conceptus. In this context, the uterine immunological responses responsible for rejection or tolerance of the conceptus are of critical importance. The aim of the present review is to summarize our current knowledge about those cellular and molecular immunological events occurring at the uterine level during pre-implantation and implantation stages of pregnancy in the pig. Advancing our understanding of the immune mechanisms involved in the success or failure of pregnancy will provide cues to develop novel strategies augmenting endometrial receptivity, finally increasing the efficiency of assisted reproductive technologies in pigs.
Kasun GodakumaraKeerthie DissanayakeMohammad Mehedi HasanSuranga. P. Kodithuwakku...
8页
查看更多>>摘要：The mammalian reproduction is a process of controlled cellular growth and development regulated by constant communication between the gametes, the subsequent embryo and the maternal system. Extracellular vesicles (EVs) are involved in these communications to a significant degree from the gamete production and maturation to fertilization, embryo development and implantation. They regulate the cellular physiology and the immune reaction to bring about a favourable environment for a successful pregnancy. Deciphering the mechanisms employed in EV-mediated embryo maternal communication could improve our knowledge in mammalian reproduction and increase the efficiency of animal breeding.
查看更多>>摘要：To date, large-scale use of multiple ovulation and embryo transfer (MOET) programmes in ovine species is limited due to unpredictable results and high costs of hormonal stimulation and treatment. Therefore, even if considered reliable, they are not fully applicable in large-scale systems. More recently, the new prospects offered by in vitro embryo production (IVEP) through collection of oocytes post-mortem or by repeated ovum pick-up from live females suggested an alternative to MOET programmes and may be more extensively used, moving from the exclusive research in the laboratory to field application. The possibility to perform oocytes recovery from juvenile lambs to obtain embryos (JIVET) offers the great advantage to significantly reduce the generation interval, speeding the rate of genetic improvement. Although in the past decades several studies implemented novel protocols to enhance embryo production in sheep, the conditions of every single stage of IVEP can significantly affect embryo yield and successful transfer into the recipients. Moreover, the recent progresses on embryo production and freezing technologies might allow wider propagation of valuable genes in small ruminants populations and may be used for constitution of flocks without risks of disease. In addition, they can give a substantial contribution in preserving endangered breeds. The new era of gene editing might offer innovative perspectives in sheep breeding, but the application of such novel techniques implies involvement of specialized operators and is limited by relatively high costs for embryo manipulation and molecular biology analysis.
ristina Ortega-FerrusolaVanesa Gómez-ArronesFrancisco E. Martín-CanoMari Cruz Gil...
11页
查看更多>>摘要：Ultrasound technology has led to new lines of research in equine reproduction, and it has helped to greatly improve clinical diagnosis and reproductive outcomes in equine practice. This review aims to discuss the potential clinical uses and new approaches of ultrasonography in equine reproduction. Doppler modalities are usually used to evaluate the vascularization of the follicles, corpus luteum (CL), and the uterus in the mare for diagnostic purposes. Inclusion of Doppler ultrasound in artificial insemination and embryo transfer programs could improve the reproductive outcome of these techniques. Better selection of recipients based on CL functionality, early pregnancy diagnosis 7-8?days postovulation of the donor before flushing or diagnosis of mares with endometritis with pathological increases of blood flow are examples of clinical applications in the mare. In the stallion, colour Doppler ultrasound has improved the diagnostic potential of B-mode ultrasound, improving the differential diagnosis of pathologies such as testicular torsion (decrease or absence of blood flow in the cord) and orchitis (increased blood flow in the cord). The incorporation of pulsed Doppler ultrasound into the reproductive evaluation of the stallion has enabled early identification of stallions with testicular dysfunction, thus allowing administration of timely treatment and subsequent improvements of the fertility prognosis for these animals. In addition, this technique has been used in the monitoring of patients undergoing medical and surgical treatments, thus verifying their efficacy. Recently, computer-assisted pixel analysis using specific software has been performed in research work in order to semi-quantitatively evaluate the vascularization (colour and power Doppler) and echotexture of different organs. These softwares are now being developed for clinical purposes, as is the case with Ecotext, a computer program developed for the evaluation of testicular echotexture, providing information on testicular functionality.
Giovanni Maria GnemmiCristina Vittoria A. MaraboliBenedetta GnemmiRoberta Saleri...
8页
查看更多>>摘要：In cattle, early detection of gestation is very important from an economic and management point of view in all types of farming. However, due to the poor efficiency of oestrous detection, it is essential to determine non-pregnant cows as early as possible, in order to minimize the inter-insemination interval, thus de facto, reducing herd open days. Direct and indirect gestation diagnostic methods have been developed with the aim of improving the reproductive performance of the herd. Today, the most accurate method for making an early diagnosis of gestation from 28 to 30?days post-insemination is B-mode ultrasound. In recent years, indirect methods have included techniques that allow non-pregnant cows to be identified with a minimum margin of error, the most widely utilized of which is the colour Doppler. This technique is rapidly becoming established for the diagnosis of non-pregnancy, which allows for the identification of non-pregnant animals earlier compared with the pregnancy diagnosis. Some limitations of this technique in dairy cow have been presented.
查看更多>>摘要：The cold-adapted bacterium Pseudomonas sp. ID1 produces the extracellular exopolysaccharide ID1 (EPS ID1) with cryoprotective activity. This study was designed to optimize the vitrification/in-straw warming protocol of in vitro-produced (IVP) blastocysts by adding EPS ID1 to the vitrification media. Day 7-expanded blastocysts were vitrified/warmed using the VitTrans device after the addition of 0 or 100?μg/mL EPS ID1 to the vitrification media. Blastocysts vitrified by the Cryotop method and fresh non-vitrified blastocysts served as controls. Outcomes were assessed in the warmed embryos in terms of survival rates and mRNA relative abundances of BAX, BCL2, GPX1, and CDX2 genes. No differences in survival rates were observed at 3 h post-warming between vitrification treatments. At 24?h post-warming, the addition of EPS prior to vitrification with the VitTrans device produced similar survival rates to Cryotop-vitrified embryos and similar hatching rates to fresh non-vitrified or Cryotop-vitrified embryos. No differences emerged in BCL2 gene expression. Lower BAX (p?<?.05) and higher GPX1 (p?<?.05) and CDX2 (p?<?.1) gene expression were observed in expanded and/or hatched blastocysts derived from VitTrans-EPS-vitrified embryos when compared to those from the non-supplemented group. In conclusion, addition of EPS not only promoted blastocyst survival and hatching after VitTrans vitrification/warming but also modified the expression of genes associated with better embryo quality.
Alejandro Gonzalez-PlazaCristiano BrulloJosep M. CambraManuela Garcia...
6页
查看更多>>摘要：The aims of this study were to investigate the effects of different equilibration times with cryoprotectants on viability and metaphase plate morphology of vitrified-warmed porcine mature oocytes (Experiment 1) and to evaluate the effects of supplementation with 10-9 M melatonin during in vitro maturation on these parameters (Experiment 2). In Experiment 1, 2,392 mature oocytes were vitrified using different equilibration times of oocytes with cryoprotectants (3, 10, 15, 20, 30, 40, 60 and 80?min). Fresh oocytes matured in vitro for 44?hr (n = 509) were used as controls. In Experiment 2, a total of 573 COCs were used. COCs were matured with 10~(-9) M melatonin supplementation or without melatonin (control). Some oocytes from each group were vitrified with a 60-min equilibration time with cryoprotectants according to the results of Experiment 1. The remaining oocytes from each maturation group were used as fresh control groups. In both experiments, oocytes were stained with 2′,7′-dichlorodihydrofuorescein diacetate and Hoechst 33342 to assess viability and metaphase plate morphology, respectively. Vitrification and warming affected (p?<?.01) oocyte viability compared with controls, which were all viable after 44?hr of IVM. In Experiment 1, the longer the equilibration time with cryoprotectants, the higher the viability. Oocytes equilibrated for 60 and 80?min had the highest (p?<?.05) viability and similar metaphase plate characteristics to the fresh control oocytes. In Experiment 2, supplementation with melatonin during in vitro maturation had no effect on oocyte viability or metaphase plate morphology of vitrified-warmed oocytes. In conclusion, under our experimental conditions, vitrified porcine mature oocytes equilibrated with cryoprotectants for 60 or 80?min exhibited the highest viability and similar metaphase plate characteristics to fresh controls. Furthermore, supplementation with 10~(-9) M melatonin during in vitro maturation had no effect on these parameters.
查看更多>>摘要：The objectives of this study were to evaluate the effect of vitrification on the DNA fragmentation rate of equine cumulus cells and to assess its relationship to oocyte in vitro maturation (IVM) after vitrification. Cumulus cells (CC) from 14 mares were recovered from COCs, previously submitted to vitrification (VIT) and IVM. The DNA fragmentation rate of the cumulus cells (CC-DF) was assessed using a chromatin dispersion test. CC-DF rates between vitrified and control COCs were statistically compared by Student's?t-test. The rates of CC-DF from control COCs were lower than in vitrified COCs. The percentage of CC-DF was not significantly different (p?>?.05) between groups of COCs able to reach metaphase II (MII > 0) and those in which oocyte maturation was not achieved (MII = 0). In conclusion, vitrification has a deleterious effect on the DNA fragmentation of equine cumulus cells; however, this parameter cannot be used as a predictor for IVM success after COCs vitrification.
Irene Sánchez-AjofrínPatricia Peris-FrauOlga García-álvarezMaría del Rocío Fernández-Santos...
4页
查看更多>>摘要：Incubation with estrous sheep serum (ESS) is required to induce in vitro capacitation of spermatozoa during in vitro fertilization of small ruminants. However, the effect of adding different serum concentrations in the fertilization media on the quality of resulting blastocysts has not yet been studied. Here, 298 sheep oocytes were co-incubated with capacitated spermatozoa with either 10% or 2% ESS. There were no differences between treatments in cleavage (10% ESS: 63.81?±?5.87% and 2% ESS: 45.31?±?5.87%) and blastocyst rates (10% ESS: 20.83?±?2.12% and 2% ESS: 15.93?±?2.12%). Nonetheless, in vitro-produced blastocysts from the 10% ESS treatment showed a higher transcript abundance of mRNAs involved in apoptosis (ITM2B and BCL2), antioxidant defence (GPX1) and growth-related imprinting (IGF2R). Our data suggest that ESS supplementation during in vitro fertilization can influence the quality of sheep embryos at later stages of development by increasing the transcription of developmentally important genes.
María Iniesta-CuerdaJi?ina HavránkováHedvika ?imná?ováOlga García-álvarez...
6页
查看更多>>摘要：Deficient sperm motility is a frequent cause of the age-related male sub-/infertility. Since the protein sirtuin 1 (SIRT1) develops anti-aging action and participates in sperm motility and ATP synthesis in mitochondria, we investigated its role in the acquisition of hyperactivated motility during capacitation. For this, the dynamics of sperm subpopulations were studied, using males of Sirt1~(+/-) heterozygous mutant mice. After 2 hr of capacitation, we observed reduced percentage of hyperactivated spermatozoa in Sirt1~(+/-) males. Interestingly, prior to capacitation, Sirt1~(+/-) spermatozoa showed higher mitochondrial superoxide levels, which could render mitochondrial injury and thereby motility defects. Accordingly, the fertilization rate of Sirt1~(+/-) males after mating was decreased. We elucidated that SIRT1 male insufficiency underlies posterior sperm defects to hyperactivate during capacitation and propose Sirt1~(+/-) males as a model for the study of the age-related infertility.
NSTL
Wiley