RESUMO
Lipopolysaccharide (LPS) derived from gram negative bacteria cell wall is known to cause ruminal acidosis and/or infectious diseases such as metritis and mastitis which has a significant negative impact on the reproductive performance. This study aimed to investigate the effect of LPS on oocyte maturation and subsequent development in vitro. Ovine cumulus oocyte complexes (COCs) were matured in a medium supplemented with 0 (control), 0.01, 0.1, 1 and 10 µg/mL LPS. Nuclear maturation, cleavage and blastocyst rate, mitochondrial membrane potential (ΔΨm), intracellular reactive oxygen species (ROS) content and changes to the transcript abundance were evaluated. In case of the maturation rate, the percentage of oocytes reaching the MII stage was lower following exposure to 10 µg/mL LPS in comparison to the control group (P < 0.05). Moreover, the blastocyst rate decreased in case of 1 and 10 µg/mL LPS when compared to the control group (P < 0.05). ROS overproduction accompanied by a decreased ΔΨm were recorded in LPS treated oocytes in comparison to the control group (P < 0.05). The 3' tag digital gene expression profiling method revealed that 7887 genes were expressed while only seven genes exhibited changes in the transcript abundance following exposure to LPS. Tripartite motif containing 25 (TRIM25), Tripartite motif containing 26 (TRIM26), Zona Pellucida glycoprotein 3 (ZP3), Family with sequence similarity 50-member A (FAM50A), Glyoxalate and hydroxy pyruvate reductase (GRHPR), NADH ubiquinase oxireductase subunit A8 (NDUFA8) were down-regulated (P < 0.05), while only Centrin 3 (CETN3) was up-regulated (P < 0.05). Our findings show that LPS has undesirable effects on the maturation competence of ovine oocytes and subsequent embryo development. In addition, the transcriptomic profiling results may shed more light on the molecular mechanisms of LPS-induced infertility in ruminants.
Assuntos
Técnicas de Maturação in Vitro de Oócitos , Lipopolissacarídeos , Animais , Blastocisto , Células do Cúmulo , Desenvolvimento Embrionário , Feminino , Técnicas de Maturação in Vitro de Oócitos/veterinária , Lipopolissacarídeos/farmacologia , Oócitos , OvinosRESUMO
This study evaluated the distribution and size of lipid droplets (LDs) in oocytes recovered from young and adult ovine ovaries. Collected oocytes were categorised on the basis of their major diameter (small (SO), 70-90 µm; medium (MO), >90-110 µm; large (LO), >110-130µm) and were stained with Nile red to detect LDs. In adult and young oocytes, a diffuse pattern distribution of LDs was dominant in all classes except adult LO and young SO and LO. Larger LDs (i.e. >3µm) were mostly present in young SO and LO, whereas smaller LDs (1-3µm) were detected in the other adult and young oocyte categories.
Assuntos
Gotículas Lipídicas/metabolismo , Oócitos/metabolismo , Ovário/metabolismo , Animais , Feminino , OvinosRESUMO
Cumulative evidence has proven the safety, feasibility and efficacy of stem cell therapy for cardiomyocyte replacement in heart failure treatment. In contrast to embryonic stem cells, induced pluripotent stem cells (iPS cells) provide a route to the production of patient-specific stem cell lines with no ethical concerns. Recent studies have revealed that myogenic transcription factors activated the expression of conserved microRNAs (miRNAs), such as mir-1, that 'fine-tuned' the output of the transcriptional networks. To introduce an efficient and applicable protocol for establishment of autologous cardiac cellular models, herein we introduced a novel protocol for induction of iPS cells into cardiomyocytes using both microRNA-1 transduction and 5'-Azacitidine treatment. Quantitative evaluation of transcription and translation of cardiac markers such as MHC-α, GATA4, FLK and troponin, demonstrated that this new direct protocol led to cardiac differentiation of iPS cells. From a clinical point of view, these results raise the possibility that administration of miRNA mimic or miRNA inhibitor therapies could increase allocation of iPS cells into the cardiac lineage. Taking all the results into account, our novel protocol provides further progress in the application of patient's own cells for more effective therapies. Moreover, such cellular models could be used in personalized drug screening.
Assuntos
Diferenciação Celular/genética , Células-Tronco Pluripotentes Induzidas/metabolismo , Modelos Biológicos , Miócitos Cardíacos/metabolismo , Azacitidina/farmacologia , Diferenciação Celular/efeitos dos fármacos , Terapia Baseada em Transplante de Células e Tecidos/métodos , Células Cultivadas , Inibidores Enzimáticos/farmacologia , Fator de Transcrição GATA4/genética , Fator de Transcrição GATA4/metabolismo , Expressão Gênica , Cardiopatias/genética , Cardiopatias/terapia , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , MicroRNAs/genética , Miócitos Cardíacos/citologia , Troponina/genética , Troponina/metabolismoRESUMO
Mesenchymal stem cells (MSCs) are multipotent stem cells found in many adult tissues, especially bone marrow (BM) and are capable of differentiation into various lineage cells such as osteoblasts, adipocytes, chondrocytes and myocytes. Moreover, MSCs can be mobilized from connective tissue into circulation and from there to damaged sites to contribute to regeneration processes. MSCs commitment and differentiation are controlled by complex activities involving signal transduction through cytokines and catecholamines. There has been an increasing interest in recent years in the neural system, functioning in the support of stem cells like MSCs. Recent efforts have indicated that the catecholamine released from neural and not neural cells could be affected characteristics of MSCs. However, there have not been review studies of most aspects involved in catecholamines-mediated functions of MSCs. Thus, in this review paper, we will try to describe the current state of catecholamines in MSCs destination and discuss strategies being used for catecholamines for migration of these cells to damaged tissues. Then, the role of the nervous system in the induction of osteogenesis, adipogenesis, chondrogenesis and myogenesis from MSCs is discussed. Recent progress in studies of signaling transduction of catecholamines in determination of the final fate of MSCs is highlighted. Hence, the knowledge of interaction between MSCs with the neural system could be applied towards the development of new diagnostic and treatment alternatives for human diseases.