WNT3A signalling pathway in buffalo (Bubalus bubalis) embryonic stem cells
Mohammad Zandi A , Musharifa Muzaffar B , Syed Mohmad Shah B , Ramakant Kaushik B , Manoj Kumar Singh B , Prabhat Palta B , Suresh Kumar Singla B , Radhey Sham Manik B and Manmohan Singh Chauhan B C
+ Author Affiliations
- Author Affiliations
A Department of Animal and Poultry Science and Fisheries, Agricultural Institute, Iranian Research Organisation for Science and Technology, Tehran 33535111, Iran.
B Embryo Biotechnology Laboratory, Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India.
C Corresponding author. Email: chauhan_abtc@rediffmail.com
Reproduction, Fertility and Development 26(4) 551-561 https://doi.org/10.1071/RD13084
Submitted: 14 November 2012 Accepted: 19 March 2013 Published: 9 May 2013
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 400 ng mL–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.
Additional keywords: differentiation, pluripotency.
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