title: WNT3A signalling pathway in buffalo (Bubalus bubalis) embryonic stem cells.
abstract: The aim of this study was to investigate the transcriptional profile and role of WNT3A signalling in maintaining buffalo embryonic stem (ES) cells in a pluripotent state and in the induction of their differentiation. ES cells were derived from embryos produced by in vitro fertilisation (iESC), parthenogenesis (pESC) and hand-made cloning (cESC). The expression of WNT3A, its receptors and intermediate signalling pathways were found to be conserved in ES cells derived from the three different sources. WNT3A was expressed in ES cells but not in embryoid bodies derived from iESC or in buffalo fetal fibroblast cells. It was revealed by real-time polymerase chain reaction analysis that following supplementation of culture medium with WNT3A (100, 200 or 400ngmL(-1)) a significant increase (P<0.05) was observed in the expression level of β-CATENIN, which indicated the activation of the canonical WNT pathway. WNT3A, in combination with exogenous fibroblast growth factor-2 and leukaemia inhibitory factor, induced proliferation of undifferentiated ES cells. Differentiation studies showed that WNT3A caused formation of scaffold-like structures and inhibition of differentiation into neuron-like cells. In conclusion, the WNT3A signalling pathway is necessary both for maintaining undifferentiated buffalo ES cells as well as for directing their differentiation.
authors: Department of Animal and Poultry Science and Fisheries, Agricultural Institute, Iranian Research Organisation for Science and Technology, Tehran 33535111, Iran.
authors: Mohammad
authors: M
authors: Zandi
authors: Embryo Biotechnology Laboratory, Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India.
authors: Musharifa
authors: M
authors: Muzaffar
authors: Embryo Biotechnology Laboratory, Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India.
authors: Syed Mohmad
authors: SM
authors: Shah
authors: Embryo Biotechnology Laboratory, Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India.
authors: Ramakant
authors: R
authors: Kaushik
authors: Embryo Biotechnology Laboratory, Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India.
authors: Manoj Kumar
authors: MK
authors: Singh
authors: Embryo Biotechnology Laboratory, Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India.
authors: Prabhat
authors: P
authors: Palta
authors: Embryo Biotechnology Laboratory, Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India.
authors: Suresh Kumar
authors: SK
authors: Singla
authors: Embryo Biotechnology Laboratory, Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India.
authors: Radhey Sham
authors: RS
authors: Manik
authors: Embryo Biotechnology Laboratory, Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India.
authors: Manmohan Singh
authors: MS
authors: Chauhan
mesh_terms: D000818:Animals; D002020:Buffaloes / Q000196:embryology / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D053605:Leukemia Inhibitory Factor / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D060490:Receptors, Wnt / Q000378:metabolism; D014158:Transcription, Genetic; D060449:Wnt Signaling Pathway; D060509:Wnt3A Protein / Q000235:genetics / Q000378:metabolism*; D051176:beta Catenin / Q000378:metabolism
chemical_list: D053605:Leukemia Inhibitory Factor; D012333:RNA, Messenger; D060490:Receptors, Wnt; D060509:Wnt3A Protein; D051176:beta Catenin; D016222:Fibroblast Growth Factor 2