Focal adhesion pathway is one of the key molecular pathways affected by suboptimal culture conditions during embryonic development. The epidermal growth factor (EGF) and hyaluronic acid (HA) are believed to be involved in the focal adhesion pathway function by regulating the adherence of the molecules to the extracellular matrix. However, regulatory and molecular mechanisms through which the EGF and HA could influence the embryo development is not clear. Therefore, this study aimed to investigate the effect of continued or stage specific supplementation of EGF and/or HA on the developmental competence and quality of bovine preimplantation embryos and the subsequent consequences on the expression and DNA methylation patterns of genes involved in the focal adhesion pathway. The results revealed that, the supplementation of EGF or HA from zygote to the blastocysts stage reduced the level of reactive oxygen species and increased hatching rate after thawing. On the other hand, HA decreased the apoptotic nuclei and increased blastocyst compared to EGF supplemented group. Gene expression and DNA methylation analysis in the resulting blastocysts indicated that, combined supplementation of EGF and HA increased the expression of genes involved in focal adhesion pathway while supplementation of EGF, HA or a combination of EGF and HA during the entire preimplantation period changed the DNA methylation patterns of genes involved in focal adhesion pathway. On the other hand, blastocysts developed in culture media supplemented with EGF + HA until the 16-cell stage exhibited higher expression level of genes involved in focal adhesion pathway compared to those supplemented after the 16-cell stage. Conversely, the DNA methylation level of candidate genes was increased in the blastocysts obtained from embryos cultured in media supplemented with EGF + HA after 16-cell stage. In conclusion, supplementation of bovine embryos with EGF and/or HA during the entire preimplantation period or in a stage specific manner altered the DNA methylation and expression patterns of candidate genes involved in the focal adhesion pathway which was in turn associated with the observed embryonic developmental competence and quality.
This study was conducted in Hadraba Valley, Halaieb and Shalateen Research Station, Desert Research Center. The objective of this study is to investigate the effectiveness of providing shade and/ or feeding probiotics as means for alleviating heat stress on Hassani goats raised in the far south of the eastern desert of Egypt indicating by thermo-cardio-respiratory responses and changes in some hemato-biochemical parameters. Four groups of mature male Hassani goats (Five animals in each) were used; group one (G1); kept unshaded without probiotics, group two (G2); kept unshaded and fed probiotics. Group three (G3) shaded and had un-supplemented probiotics; while group four (G4); shaded and fed probiotics. Meteorological data in terms of radiant ambient temperature (RAT), ambient temperature (AT) and relative humidity (RH) and thermocardio-respiratory responses (rectal; RT, skin; ST and coat temperatures; CT and respiration; RR and heart rates; HR) were recorded twice daily at 08.00 and 14.00 hr. Hemoglobin (Hb), packed cell volume (PCV), mean of corpuscular hemoglobin concentration (MCHC), plasma total cholesterol (TC), glucose (GLU), total proteins (TP), albumin (A), globulin (G), A/G ratio, alanine amino transferase (ALT) and aspartate amino transferase (AST) were determined.The results revealed that providing probiotics relieved the burden of heat stress as indicated by reducing (P<0.01) the thermo-cardio-respiratory responses and increasing (P<0.05) TP and G. Moreover, probiotics caused a non-significant elevation of mean values of PCV, HB, MCHC, A, GLU and reduction of A/G ratio, TC and AST and ALT. on the other hand, Shading resulted in significant (P<0.01) reductions in RT, CT and RR and non-significant reductions in ST, HR, TP, A, G, AST, ALT. Furthermore, shading caused significant (P<0.05) increases in PCV and MCHC and non-significant increases in Hb, TC and GLU. The improvement in thermo-cardio-respiratory responses and hemobiochemical parameters due to probiotics were more pronounced than those of shading. However, the best benefits were obtained of the group provided the two treatments together.It could be concluded that providing shade and/ or probiotics for heat stressed animals in such remote region would improve their heat tolerance to the severe hot conditions prevailing in this region.
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