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Ivona Travnickova Pavlina Hulinska Svatava Kubickova Katerina Hanzalova Bartozs Kempisty Lucie Nemcova Marie Machatkova 《Reproduction in domestic animals》2021,56(6):864-871
The study investigated the effects of sperm sorting, capacitation treatment and co-cultivation on sexed bovine in vitro embryo production. The effect of treatment and co-culture on production of embryos of the preferred sex from unsorted sperm was also studied. Sperm from five breeding bulls was used for fertilization of mature oocytes as follows : Experiment 1, sorted and unsorted sperm (bulls A-E) treated only with heparin in standard co-cultures; Experiment 2, sorted sperm (bulls A-E) treated with heparin-PHE (penicillamine, hypotaurine, and epinephrine) or heparin-caffeine in drop co-cultures; and Experiment 3, unsorted sperm (bull E) treated with either heparin-PHE or heparin-caffeine in both standard and drop co-cultures. In all bulls, treatment with heparin resulted in significantly (p < .05) reduced cleavage and blastocyst rates from sorted sperm, as compared with those from unsorted sperm. In bulls A, B, D and E, treatment of sorted sperm with heparin-PHE in drops significantly increased the blastocyst rate (p < .05). In unsorted sperm of bull E, heparin-PHE treatment in drops resulted in the XX/XY sex ratio inverse to that obtained by heparin-caffeine treatment in standard co-cultures (32.3%/67.7% and 66.7%/33.3%, respectively). In conclusion, the treatment of sorted sperm with heparin-PHE in modified drop co-cultures can be recommended for production of in vitro sexed embryos. The use of unsorted sperm for production of embryos of the preferred sex by selected capacitation treatment and co-culture can be the method of choice in bulls with low IVF yields from sorted sperm. 相似文献
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Analysis of integrins and vascular endothelial growth factor isoforms mRNA expression in the canine uterus during perimplantation period 总被引：1，自引：0，他引：1
Bukowska D Kempisty B Jackowska M Woźna M Antosik P Piotrowska H Jaśkowski JM 《Polish journal of veterinary sciences》2011,14(2):253-258
Integrins are the major receptors within the extracellular matrix (ECM) that mediate several functions connected with cell life and metabolism, such as cell adhesion, migration, cytoskeletal organization, proliferation, survival, and differentiation. A vascular endothelial growth factor (VEGF) is one of the most important angiogenic factors. It has been suggested that the expression of this gene may play crucial physiological roles in reproductive organs. All investigated endometrial tissues were isolated on day 10-12 after mating. Control bitches, used in this study, were in metestrus, which was determined according to the vaginal cytology and progesterone level in blood. Early pregnancy was verified by flushing the uterine horns with PBS. Total RNA was isolated from the bitches endometrium by means of the Chomczyński and Sacchi method, treated by DNase I, and reverse-transcribed into cDNA. A quantitative analysis of integrins alpha2b, beta2 and beta3, VEGF 164, 182 and 188 cDNA was performed by RT-PCR. In results we have shown an increased expression of all investigated genes (integrins alpha2b, beta2 and beta3, VEGF 164, 182, and 188) in pregnant bitches uterus as compared to non-pregnant females (P < 0.001). Our results indicated that the expression of genes encoding integrins and vascular endothelial growth factors is different in relation to the time of the embryo implantation and it is increased in the first period of this process. This may be associated with the induction of specific mechanisms responsible for receptivity of uterus following the embryo attachment. In addition, all of investigated genes are up-regulated in a pregnancy-specific manner and the increased expression of these genes may regulate the uterus function during the implantation of canine embryos. 相似文献
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Bukowska D Kempisty B Piotrowska H Zawierucha P Brussow KP Jaśkowski JM Nowicki M 《Polish journal of veterinary sciences》2012,15(1):199-205
The maturation of oocytes is one of the most important steps determining their developmental competence. Due to the low percentage of oocytes of bitches that reach the MII stage, searching for reagents that may stimulate the growth and maturation of oocytes is still present in this species of mammals. The most important media supplements include gonadotropins (LH, FSH, hCG), growth factors (IGF, TGF, EGF, FGF), progesterone and follicular fluid. It is suggested that the supplement of EGF, and/or follicular cells may have an important influence on the percentage of cells that reach the MII stage. Despite plenty of research based on the improvement of bitch oocytes in vitro culture, the results obtained are still unsatisfactory. Moreover, in the long stages of canine oocytes maturation many molecular and morphological modifications (including changes in mitochondria structure and configuration in the cytoplasm) are involved. In this article, the influence of selected media supplements on the efficiency of bitch oocytes in vitro maturation was described. The molecular and morphological modifications during canine oocytes maturation were also considered in the text. 相似文献
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Energy Status Characteristics of Porcine Oocytes During In Vitro Maturation is Influenced by Their Meiotic Competence 
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下载免费PDF全文 I Milakovic M Jeseta S Hanulakova D Knitlova K Hanzalova P Hulinska L Machal B Kempisty P Antosik M Machatkova 《Reproduction in domestic animals》2015,50(5):812-819
The characteristics of energy status in porcine oocytes as related to their meiotic competence and in vitro maturation were studied. Cycling pubertal gilts in the early luteal to early follicular phases of the ovarian cycle were used as oocyte donors. The oocytes recovered from medium (MF) or small follicles (SF) were considered meiotically more or less competent, respectively. A half of oocytes from each category was matured by the standard protocol. The oocytes were examined before or after maturation by confocal microscopy, a bioluminescent cell assay and Western blotting. Four experiments, each in triplicate, were performed to assess both SF and MF oocytes in terms of metabolic units formed by mitochondria and lipids, ATP and lipid consumption and lipid droplets with adipose differentiation‐related protein (ADRP) expression. The proportion of oocytes with metabolic units, the mean ATP content and the number of lipid droplets per oocyte, and the relative number of lipid droplets with ADRP expression were significantly higher in the MF compared to SF oocytes before maturation. On the other hand, after maturation, there was an increase in the proportion of oocytes with metabolic units and the relative number of lipid droplets with ADRP expression in the SF compared to MF oocytes. In conclusion, specific differences in energy characteristics between porcine oocytes with different meiotic competence were found. Meiotically more competent oocytes are more advanced in terms of energy reserves before maturation, while meiotically less competent oocytes are more active in replenishing energy stores during maturation. 相似文献
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