title: Combination of melatonin and Wnt-4 promotes neural cell differentiation in bovine amniotic epithelial cells and recovery from spinal cord injury.
abstract: Although melatonin has been shown to exhibit a wide variety of biological functions, its effects on promoting differentiation of neural cells remain unknown. Wnt signaling mediates major developmental processes during embryogenesis and regulates maintenance, self-renewal, and differentiation of adult mammalian stem cells. However, the role of the noncanonical Wnt pathway during neurogenesis remains poorly understood. In this study, the amniotic epithelial cells ( AECs) were isolated from bovine amnion and incubated with various melatonin concentrations (0.01, 0.1, 1, 10, or 100 μm) and 5 × 10(-5) m all-trans retinoic acid (RA) for screening optimum culture medium of neural differentiation, compared with each groups, 1 μm melatonin and 5 × 10(-5) m RA were selected to induce neural differentiation of AECs, and then siMT1, siMT2, oWnt-4, and siWnt-4 were expressed in AECs to research role of these genes in neural differentiation. Efficiency of neural differentiation was evaluated after expressed above genes using flow cytometry. Cell function of neural cells was demonstrated in vivo using spinal cord injury model after cell transplantation, and damage repair of spinal cord was assessed using cell tracking and Basso, Beattie, Bresnahan Locomotor Rating Scale scores. Results demonstrated that melatonin stimulated melatonin receptor 1, which subsequently increased bovine amniotic epithelial cell vitality and promoted differentiation into neural cells. This took place through cooperation with Wnt-4. Additionally, following cotreatment with melatonin and Wnt-4, neurogenesis gene expression was significantly altered. Furthermore, single inhibition of melatonin receptor 1 or Wnt-4 expression decreased expression of neurogenesis-related genes, and bovine amniotic epithelial cell-derived neural cells were successfully colonized into injured spinal cord, which suggested participation in tissue repair.
authors.affiliation: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.|College of Wildlife Resources, Northeast Forestry University, Harbin, China.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Chunyu
authors.initials: C
authors.lastname: Bai
authors.affiliation: College of Wildlife Resources, Northeast Forestry University, Harbin, China.
authors.forename: Dong
authors.initials: D
authors.lastname: Zheng
authors.affiliation: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Changli
authors.initials: C
authors.lastname: Li
authors.affiliation: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Wenxiu
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Mei
authors.initials: M
authors.lastname: Li
authors.affiliation: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Weijun
authors.initials: W
authors.lastname: Guan
authors.affiliation: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Yuehui
authors.initials: Y
authors.lastname: Ma
mesh_terms: D000650:Amnion / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D002454:Cell Differentiation / Q000187:drug effects*; D004847:Epithelial Cells / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D008550:Melatonin / Q000494:pharmacology*; D055495:Neurogenesis / Q000187:drug effects; D013119:Spinal Cord Injuries / Q000378:metabolism*; D058630:Spinal Cord Regeneration / Q000187:drug effects*; D060528:Wnt4 Protein / Q000378:metabolism*
chemical_list: D060528:Wnt4 Protein; D008550:Melatonin
keywords: Wnt-4; amniotic epithelial cells; bovine; melatonin; neural differentiation; neuroectodermal marker