title: Melatonin promotes development of haploid germ cells from early developing spermatogenic cells of Suffolk sheep under in vitro condition.
abstract: Promotion of spermatogonial stem cell (SSC) differentiation into functional sperms under in vitro conditions is a great challenge for reproductive physiologists. In this study, we observed that melatonin (10(-7) M) supplementation significantly enhanced the cultured SSCs differentiation into haploid germ cells. This was confirmed by the expression of sperm special protein, acrosin. The rate of SSCs differentiation into sperm with melatonin supplementation was 11.85 ± 0.93% which was twofold higher than that in the control. The level of testosterone, the transcriptions of luteinizing hormone receptor (LHR), and the steroidogenic acute regulatory protein (StAR) were upregulated with melatonin treatment. At the early stage of SSCs culture, melatonin suppressed the level of cAMP, while at the later stage, it promoted cAMP production. The similar pattern was observed in testosterone content. Expressions for marker genes of meiosis anaphase, Dnmt3a, and Bcl-2 were upregulated by melatonin. In contrast, Bax expression was downregulated. Importantly, the in vitro-generated sperms were functional and they were capable to fertilize oocytes. These fertilized oocytes have successfully developed to the blastula stage.
authors.affiliation: State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Shou-Long
authors.initials: SL
authors.lastname: Deng
authors.affiliation: State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Su-Ren
authors.initials: SR
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Zhi-Peng
authors.initials: ZP
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Ji-Xin
authors.initials: JX
authors.lastname: Tang
authors.affiliation: State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.affiliation: State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Xiu-Xia
authors.initials: XX
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Jin-Mei
authors.initials: JM
authors.lastname: Cheng
authors.affiliation: State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Jin
authors.affiliation: State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Xiao-Yu
authors.initials: XY
authors.lastname: Li
authors.affiliation: National key Lab of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, P.R. China.
authors.forename: Bao-Lu
authors.initials: BL
authors.lastname: Zhang
authors.affiliation: National key Lab of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, P.R. China.
authors.forename: Kun
authors.initials: K
authors.lastname: Yu
authors.affiliation: Laboratory of Animal Genetics and Breeding, College of Animal Science and Technology, China Agricultural University, Beijing, P.R. China.
authors.forename: Zheng-Xing
authors.initials: ZX
authors.lastname: Lian
authors.affiliation: Laboratory of Animal Genetics and Breeding, College of Animal Science and Technology, China Agricultural University, Beijing, P.R. China.
authors.forename: Guo-Shi
authors.initials: GS
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Yi-Xun
authors.initials: YX
authors.lastname: Liu
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008297:Male; D008550:Melatonin / Q000494:pharmacology*; D060888:Real-Time Polymerase Chain Reaction; D012756:Sheep; D020554:Sperm Injections, Intracytoplasmic / Q000187:drug effects; D013091:Spermatogenesis / Q000187:drug effects*; D013094:Spermatozoa / Q000166:cytology* / Q000187:drug effects*; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects
chemical_list: D000975:Antioxidants; D008550:Melatonin
keywords: Suffolk lambs; haploid germ cells; in vitro SSCs culture system; melatonin