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Nanos2 promotes differentiation of chicken ( Gallus gallus ) embryonic stem cells to male germ cells
Author(s) -
Zhang Wenhui,
Bi Yulin,
Wang Yingjie,
Li Dong,
He Nana,
Wang Man,
Jin Jing,
Zuo Qisheng,
Zhang Yani,
Li Bichun
Publication year - 2018
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.26528
Subject(s) - biology , microbiology and biotechnology , knockout mouse , germ cell , embryonic stem cell , flow cytometry , gene , genetics
Nanos2 is an evolutionarily conserved RNA‐binding protein containing 2 CCHC‐type zinc finger motives. Here, we report that Nanos2 is strongly expressed in the testis compared to other tissues in chicken ( Gallus gallus ). Overexpression and knockout plasmid vectors were constructed, and in‐vitro Cas9/gRNA digestion and T7 endonuclease I (T7E1) assay indicated that Nanos2 ‐g1 possessed the highest knockout activity. In vitro and in vivo, Nanos2 overexpression accelerated the production of embryoid bodies (EBs) and SSC‐like cells and promoted cvh , c‐kit , and integrin α6 expression. Immunofluorescence staining, periodic acid schiff (PAS) and flow cytometry (FCM) assay showed that primordial germ cells (PGCs) and spermatogonial stem cells (SSCs) formation were significantly promoted. On the contrary, Nanos2 knockout delayed the production of EBs and SSC‐like cells and correspondingly reduced cvh , c‐kit , and integrin α6 expression. Simultaneously, the quantity of PGCs and SSCs was blocked. Collectively, these results uncovered a novel function of Nanos2 involved in chicken male germ cell differentiation, where it acts as a facilitator.