miR-544 Regulates Dairy Goat Male Germline Stem Cell Self-Renewal via Targeting PLZF

• Published in: Journal of cellular biochemistry Vol. 116; no. 10; pp. 2155 - 2165
• Main Authors: Song, Wencong, Mu, Hailong, Wu, Jiang, Liao, Mingzhi, Zhu, Haijing, Zheng, Liming, He, Xin, Niu, Bowen, Zhai, Yuanxin, Bai, Chunling, Lei, Anmin, Li, Guangpeng, Hua, Jinlian
• Format: Journal Article
• Language: English
• Published: United States Blackwell Publishing Ltd 01.10.2015; Wiley Subscription Services, Inc
• Subjects: Animals; DAIRY GOAT; Germ Cells - growth & development; Germ Cells - metabolism; Goats - genetics; Kruppel-Like Transcription Factors - biosynthesis; Kruppel-Like Transcription Factors - genetics; Male; MALE GERMLINE STEM CELLS (mGSCs); MicroRNAs - biosynthesis; MicroRNAs - genetics; miR-544; PLZF; Promyelocytic Leukemia Zinc Finger Protein; Signal Transduction; Spermatogenesis - genetics; Stem Cells; MALE GERMLINE STEM CELLS (mGSCs); DAIRY GOAT; miR-544; PLZF
• ISSN: 0730-2312; 1097-4644
• DOI: 10.1002/jcb.25172

ABSTRACT The balance between the self‐renewal and differentiation of male germline stem cells (mGSCs) is critical for the initiation and maintenance of mammalian spermatogenesis. The promyelocytic leukemia zinc finger (PLZF), a zinc finger protein, is a critical factor for maintaining the self‐renewal of mGSCs, so, evaluation of the PLZF pathway in mGSCs may provide a deeper insight into mammalian spermatogenesis. miRNA was also an important regulating factor for the self‐renewal and differentiation of mGSCs; however, there is currently no data indicating that which miRNA regulate the self‐renewal and differentiation of mGSCs via PLZF. Here, we predicted the prospective miRNA targeting to PLZF using the online Bioinformatics database‐Targetscan, and performed an analysis of the dual‐luciferase recombinant vector, psiCHCEKTM‐2‐PLZF‐3′UTR. miR‐544 mimics (miR‐544m), miR‐544 inhibitors (miR‐544i), Control (NC, scrambled oligonucleotides transfection), pPLZF‐IRES2‐EGFP or PLZF siRNA were transfected into mGSCs; the cells proliferation was evaluated by BRDU incorporation assay and flow cytometry, and the mGSC marker, GFRa1, PLZF, KIT, DAZL, and VASA expression were analyzed by RT‐qPCR, immunofluorescence and Western blot. The results showed that miR‐544 regulates dairy goat male germline stem cell self‐renewal via targeting PLZF. Our study identifies a new regulatory pathway for PLZF and expands upon the PLZF regulatory network in mGSCs. J. Cell. Biochem. 116: 2155–2165, 2015. © 2015 Wiley Periodicals, Inc.