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Regulation of Hedgehog Signaling in Chicken Embryonic Stem Cells Differentiation Into Male Germ Cells ( Gallus )
Author(s) -
Chen Hao,
Zuo Qisheng,
Wang Yinjie,
Ahmed Mahmoud F.,
Jin Kai,
Song Jiuzhou,
Zhang Yani,
Li Bichun
Publication year - 2017
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.25796
Subject(s) - hedgehog signaling pathway , biology , microbiology and biotechnology , embryonic stem cell , hedgehog , signal transduction , stem cell , cellular differentiation , indian hedgehog , integrin , retinoic acid , cell , genetics , cell culture , gene
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.