title: Inhibition of FGF Signalling Pathway Augments the Expression of Pluripotency and Trophoblast Lineage Marker Genes in Porcine Parthenogenetic Blastocyst.
abstract: The consistent failure to isolate bona fide pluripotent cell lines from livestock indicates that the underlying mechanisms of early lineage specification are poorly defined. Unlike other species, the contrivances of segregation have been comprehensively studied in the mouse. In mouse, FGF/MAPK signalling pathway dictates the segregation of hypoblast (primitive endoderm). However, it is not evident whether this mechanism is also conserved in livestock. Here, in this study, we examined the roles of FGF/MAP kinase signalling pathways in porcine parthenogenetic embryos during the early development. Porcine parthenogenetic embryos were cultured in the medium addition with FGFR inhibitor BGJ398 (10 μm) or DEMOS. Pluripotency- and lineage-related gene expressions in the early porcine embryos were determined. Compared to control, total cell numbers on day 7 were significantly higher (55 ± 5.96 vs 47 ± 1.97, p < 0.05) in embryos cultured in the presence of BGJ398, but had no significant effect on the rate of blastocyst development (47% vs 44%, p > 0.05). Nonetheless, BGJ398 treatment significantly augmented the expression of pluripotency and trophoblast marker genes (SOX2, OCT4, KLF4 and CDX2), but did not significantly change the expression of NANOG and hypoblast marker gene (GATA4). Furthermore, the addition of FGF signalling agonist (FGF2) during the embryo development significantly decreased the expression of pluripotency and trophoblast marker genes (SOX2, NANOG, KLF4 and CDX2), but no significant effect on the expression of OCT4 and GATA4 was observed. Here, we exhibit that inhibition of FGF signalling could improve the quality of the porcine embryo and escalate the chance to capture pluripotency. Besides, it also promotes the trophoblast development of porcine parthenogenetic embryo. In addition, the data suggested that FGF signalling pathway is dispensable for the segregation of hypoblast and epiblast lineages in porcine embryo during the early development.
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, China.|School of Animal Science and Technology, Guangxi University, Nanning, China.
authors.forename: L Y
authors.initials: LY
authors.lastname: Li
authors.affiliation: School of Animal Science and Technology, Guangxi University, Nanning, China.
authors.forename: M M
authors.initials: MM
authors.lastname: Li
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, China.|School of Animal Science and Technology, Guangxi University, Nanning, China.
authors.forename: S F
authors.initials: SF
authors.lastname: Yang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, China.|School of Animal Science and Technology, Guangxi University, Nanning, China.
authors.forename: J
authors.initials: J
authors.lastname: Zhang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, China.|School of Animal Science and Technology, Guangxi University, Nanning, China.
authors.forename: Z
authors.initials: Z
authors.lastname: Li
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, China.|School of Animal Science and Technology, Guangxi University, Nanning, China.
authors.forename: H
authors.initials: H
authors.lastname: Zhang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, China.|School of Animal Science and Technology, Guangxi University, Nanning, China.
authors.forename: L
authors.initials: L
authors.lastname: Zhu
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, China.|School of Animal Science and Technology, Guangxi University, Nanning, China.
authors.forename: X
authors.initials: X
authors.lastname: Zhu
authors.affiliation: Centre of Biotechnology, Nehru Science Centre, University of Allahabad, Allahabad, India.
authors.forename: V
authors.initials: V
authors.lastname: Verma
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, China. qyliu2002@126.com.|School of Animal Science and Technology, Guangxi University, Nanning, China. qyliu2002@126.com.
authors.forename: Q
authors.initials: Q
authors.lastname: Liu
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, China. deshunshi@gxu.edu.cn.|School of Animal Science and Technology, Guangxi University, Nanning, China. deshunshi@gxu.edu.cn.
authors.forename: D
authors.initials: D
authors.lastname: Shi
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, China. arihuangben@yahoo.com, benhuang@gxu.edu.cn.|School of Animal Science and Technology, Guangxi University, Nanning, China. arihuangben@yahoo.com, benhuang@gxu.edu.cn.
authors.forename: B
authors.initials: B
authors.lastname: Huang
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D005346:Fibroblast Growth Factors / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D005819:Genetic Markers; D000090062:Kruppel-Like Factor 4; D010312:Parthenogenesis / Q000502:physiology*; D039904:Pluripotent Stem Cells / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology; D013552:Swine; D014327:Trophoblasts / Q000502:physiology*
chemical_list: D005819:Genetic Markers; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D005346:Fibroblast Growth Factors