Expression pattern of Ngn3 in dairy goat testis and its function in promoting meiosis by upregulating Stra8.
OBJECTIVE
Ngn3 is a typical transcription factor and marker of differentiating spermatogonial stem cells (SSCs) in mouse, belonging to the basic helix-loop-helix (bHLH) family. Its gene is specifically expressed in A type spermatogonia in mouse testis, thus plays a critical role in controlling differentiation of SSCs. However, roles of Ngn3 and its protein in dairy goat testis remain unknown.


METHODS
Testis development and expression patterns of Ngn3 were analysed by immunofluorescence and quantitative reverse transcription-polymerase chain reaction (QRT-PCR) in the dairy goat. Furthermore, effects of its overexpression on male germline stem cells (mGSCs) were evaluated by QRT-PCR, immunofluorescence, luciferase reporter assay and western blotting.


RESULTS
Revealed that Ngn3 was expressed more highly during puberty and in the adult than in testis of other ages. Overexpression of Ngn3 promoted expression of meiosis-related gene Stra8 and stem-cell differentiation marker CD117, but suppressed expression of Plzf, a classical marker of SSCs. Furthermore, Ngn3 did not promote expression of Stra8 directly as shown in transcription and translation levels detected by luciferase reporter assay and western blotting.


CONCLUSIONS
Taken together, these results suggest that Ngn3 plays an important role in spermatogenesis and that overexpression of Ngn3 can promote meiosis in testis of the dairy goat.
College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Key Lab for Animal Biotechnology of Agriculture Ministry of China, Northwest A&F University, Yangling, Shaanxi, 712100, China.
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F
Tang
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Yao
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H
Zhu
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H
Mu
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Z
Niu
M
M
Yu
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Yang
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S
Peng
G
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Li
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J
Hua
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000367:Age Factors; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics*; D005786:Gene Expression Regulation / Q000502:physiology; D006041:Goats / Q000235:genetics*; D008297:Male; D008540:Meiosis / Q000502:physiology*; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics*; D012741:Sexual Maturation / Q000502:physiology; D013045:Species Specificity; D013093:Spermatogonia / Q000166:cytology / Q000502:physiology*; D013737:Testis / Q000166:cytology / Q000502:physiology*; D015533:Transcriptional Activation / Q000502:physiology; D015854:Up-Regulation / Q000235:genetics
D048868:Adaptor Proteins, Signal Transducing; D051792:Basic Helix-Loop-Helix Proteins; D009419:Nerve Tissue Proteins; C407737:Neurog3 protein, mouse; C102019:Stra8 protein, mouse