title: Regulation of Hedgehog Signaling in Chicken Embryonic Stem Cells Differentiation Into Male Germ Cells (Gallus).
abstract: The study aims to analyze the key signaling pathways in regulating the process of embryonic stem cells (ESCs) differentiation into spermatogonial stem cells (SSCs). Based on RNA Sequencing result, we further explored the specific regulating mechanisms of Hedgehog (HH) signaling in this process. HH signaling was found to be a crucial signaling pathway participating in the differentiation process of ESCs to SSCs. In Retinoic acid (RA) induced in vitro differentiation assay, the expression of two germ cell marker genes, integrin α6, and integrin β1, was observed to significantly increase, while it decreased dramatically when IHH was knocked down. Fluorescence activated cell sorting analysis showed that the proportion of integrin α6+and integrin β1+cells in the RA group was significantly higher than that in the RA + siRNA- Indian Hedgehog (IHH) group. In in vivo situations, siRNA-IHH could stably express in fertilized chicken embryos and significantly down-regulate the IHH expression. With real-time quantitative PCR and western blot, we identified that integrin α6 and integrin β1 expression was significantly suppressed along with IHH inhibition in vivo. We concluded that Hedgehog signaling pathway positively regulates the differentiation of ESCs to male germ cells through signal transduction by IHH. J. Cell. Biochem. 118: 1379-1386, 2017. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Orthopaedics, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, 215006, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Chen
authors.affiliation: Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, College of Animal Science and Technology,, Yangzhou University, Yangzhou, 225009, China.
authors.forename: Qisheng
authors.initials: Q
authors.lastname: Zuo
authors.affiliation: Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, College of Animal Science and Technology,, Yangzhou University, Yangzhou, 225009, China.
authors.forename: Yinjie
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Suez Canal University, Ismailia, 41522, Egypt.
authors.forename: Mahmoud F
authors.initials: MF
authors.lastname: Ahmed
authors.affiliation: Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, College of Animal Science and Technology,, Yangzhou University, Yangzhou, 225009, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Jin
authors.affiliation: Animal and Avian Sciences, University of Maryland, College Park, Maryland, 20741.
authors.forename: Jiuzhou
authors.initials: J
authors.lastname: Song
authors.affiliation: Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, College of Animal Science and Technology,, Yangzhou University, Yangzhou, 225009, China.
authors.forename: Yani
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, College of Animal Science and Technology,, Yangzhou University, Yangzhou, 225009, China.
authors.forename: Bichun
authors.initials: B
authors.lastname: Li
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002645:Chickens; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods*; D018507:Gene Expression Regulation, Developmental; D053823:Hedgehog Proteins / Q000235:genetics* / Q000378:metabolism; D039503:Integrin alpha6 / Q000235:genetics / Q000378:metabolism; D019012:Integrin beta1 / Q000235:genetics / Q000378:metabolism; D008297:Male; D017423:Sequence Analysis, RNA / Q000379:methods*; D015398:Signal Transduction; D013093:Spermatogonia / Q000166:cytology* / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D053823:Hedgehog Proteins; D039503:Integrin alpha6; D019012:Integrin beta1; D014212:Tretinoin
keywords: ESCs; HEDGEHOG SIGNALING PATHWAY; PGCs; RNA-seq; SSCs