title: Toxoplasma gondii inhibits differentiation of C17.2 neural stem cells through Wnt/β-catenin signaling pathway.
abstract: Toxoplasma gondii is a major cause of congenital brain disease. T. gondii infection in the developing fetus frequently results in major neural developmental damage; however, the effects of the parasite infection on the neural stem cells, the key players in fetal brain development, still remain elusive. This study is aiming to explore the role of T. gondii infection on differentiation of neural stem cells (NSCs) and elucidate the underlying molecular mechanisms that regulate the inhibited differentiation of NSCs induced by the infection. Using a differentiation medium, i.e. , DMEM: F12 (1:1 mixture) supplemented with 2% N2, C17.2 neural stem cells (NSCs) were able to differentiate to neurons and astrocytes, respectively evidenced by immunofluorescence staining of differentiation markers including βIII-tubulin and glial fibrillary acidic protein (GFAP). After 5-day culture in the differentiation medium, the excreted-secreted antigens of T. gondii (Tg-ESAs) significantly down-regulated the protein levels of βIII-tubulin and GFAP in C17.2 NSCs in a dose-dependent manner. The protein level of β-catenin in the nucleus of C17.2 cells treated with both wnt3a (a key activator for Wnt/β-catenin signaling pathway) and Tg-ESAs was significantly lower than that in the cells treated with only wnt3a, but significantly higher than that in the cells treated with only Tg-ESAs. In conclusion, the ESAs of T. gondii RH blocked the differentiation of C17.2 NCSs and downregulated the expression of β-catenin, an essential component of Wnt/β-catenin signaling pathway. The findings suggest a new mechanism underlying the neuropathogenesis induced by T. gondii infection, i.e. inhibition of the differentiation of NSCs via blockade of Wnt/β-catenin signaling pathway, such as downregulation of β-catenin expression by the parasite ESAs.
authors.affiliation: Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, Anhui Medical University, Hefei 230032, PR China.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Gan
authors.affiliation: Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, Anhui Medical University, Hefei 230032, PR China.
authors.forename: Xian
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, Anhui Medical University, Hefei 230032, PR China.
authors.forename: Zhengyang
authors.initials: Z
authors.lastname: Cheng
authors.affiliation: Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, Anhui Medical University, Hefei 230032, PR China.
authors.forename: Lingzhi
authors.initials: L
authors.lastname: Chen
authors.affiliation: Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, Anhui Medical University, Hefei 230032, PR China.
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Ding
authors.affiliation: Department of Biochemistry, Anhui Medical University, Hefei 230032, PR China.
authors.forename: Jian
authors.initials: J
authors.lastname: Du
authors.affiliation: Department of Health Inspection and Quarantine, School of Public Health, Anhui Medical University, Hefei 230032, PR China.
authors.forename: Yihong
authors.initials: Y
authors.lastname: Cai
authors.affiliation: Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, Anhui Medical University, Hefei 230032, PR China.
authors.forename: Qingli
authors.initials: Q
authors.lastname: Luo
authors.affiliation: Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, Anhui Medical University, Hefei 230032, PR China.
authors.forename: Jilong
authors.initials: J
authors.lastname: Shen
authors.affiliation: School of Life Sciences, Anhui University, Hefei 230601, PR China. Electronic address: yongzhong00@126.com.
authors.forename: Yongzhong
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, Anhui Medical University, Hefei 230032, PR China. Electronic address: lilyyu33@126.com.
authors.forename: Li
authors.initials: L
authors.lastname: Yu
mesh_terms: D000818:Animals; D000941:Antigens / Q000737:chemistry; D002454:Cell Differentiation; D002467:Cell Nucleus / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D015536:Down-Regulation; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D058953:Neural Stem Cells / Q000166:cytology* / Q000469:parasitology*; D009474:Neurons / Q000378:metabolism; D014122:Toxoplasma; D051153:Wnt Proteins / Q000378:metabolism*; D060449:Wnt Signaling Pathway; D051176:beta Catenin / Q000378:metabolism*
chemical_list: D000941:Antigens; D005904:Glial Fibrillary Acidic Protein; D051153:Wnt Proteins; D051176:beta Catenin; C528900:glial fibrillary astrocytic protein, mouse; D049452:Green Fluorescent Proteins
keywords: C17.2 neural stem cells; Differentiation; Excreted-secreted antigens; Toxoplasma gondii; β-Catenin