NODAL secreted by male germ cells regulates the proliferation and function of human Sertoli cells from obstructive azoospermia and nonobstructive azoospermia patients.
This study was designed to explore the regulatory effects of male germ cell secreting factor NODAL on Sertoli cell fate decisions from obstructive azoospermia (OA) and nonobstructive azoospermia (NOA) patients. Human Sertoli cells and male germ cells were isolated using two-step enzymatic digestion and SATPUT from testes of azoospermia patients. Expression of NODAL and its multiple receptors in human Sertoli cells and male germ cells were characterized by reverse transcription-polymerase chain reaction (RT-PCR) and immunochemistry. Human recombinant NODAL and its receptor inhibitor SB431542 were employed to probe their effect on the proliferation of Sertoli cells using the CCK-8 assay. Quantitative PCR and Western blots were utilized to assess the expression of Sertoli cell functional genes and proteins. NODAL was found to be expressed in male germ cells but not in Sertoli cells, whereas its receptors ALK4, ALK7, and ACTR-IIB were detected in Sertoli cells and germ cells, suggesting that NODAL plays a regulatory role in Sertoli cells and germ cells via a paracrine and autocrine pathway, respectively. Human recombinant NODAL could promote the proliferation of human Sertoli cells. The expression of cell cycle regulators, including CYCLIN A, CYCLIN D1 and CYCLIN E, was not remarkably affected by NODAL signaling. NODAL enhanced the expression of essential growth factors, including GDNF, SCF, and BMP4, whereas SB431542 decreased their levels. There was not homogeneity of genes changes by NODAL treatment in Sertoli cells from OA and Sertoli cell-only syndrome (SCO) patients. Collectively, this study demonstrates that NODAL produced by human male germ cells regulates proliferation and numerous gene expression of Sertoli cells.
Ru-Hui
RH
Tian
Shi
S
Yang
Zi-Jue
ZJ
Zhu
Jun-Long
JL
Wang
Yun
Y
Liu
Chencheng
C
Yao
Meng
M
Ma
Ying
Y
Guo
Qingqing
Q
Yuan
Yanan
Y
Hai
Yi-Ran
YR
Huang
Department of Urology, Institute of Andrology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University; State Key Laboratory of Oncogenes and Related Genes, Renji Med-X Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai Institute of Cancer, Shanghai 200127, China.
Zuping
Z
He
Department of Urology, Institute of Andrology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China.
Zheng
Z
Li
D030201:Activin Receptors, Type I / Q000378:metabolism; D030301:Activin Receptors, Type II / Q000378:metabolism; D000328:Adult; D019898:Autocrine Communication; D053713:Azoospermia / Q000378:metabolism*; D001549:Benzamides / Q000494:pharmacology; D015153:Blotting, Western; D055415:Bone Morphogenetic Protein 4 / Q000187:drug effects / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects*; D019925:Cyclin A / Q000187:drug effects / Q000378:metabolism; D019938:Cyclin D1 / Q000187:drug effects / Q000378:metabolism; D019927:Cyclin E / Q000187:drug effects / Q000378:metabolism; D004149:Dioxoles / Q000494:pharmacology; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D055457:Nodal Protein / Q000378:metabolism / Q000494:pharmacology*; D019899:Paracrine Communication; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012708:Sertoli Cells / Q000187:drug effects*; D013094:Spermatozoa / Q000378:metabolism*; D019089:Stem Cell Factor / Q000187:drug effects / Q000378:metabolism
C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; C525809:BMP4 protein, human; D001549:Benzamides; D055415:Bone Morphogenetic Protein 4; D019925:Cyclin A; D019927:Cyclin E; D004149:Dioxoles; D051100:Glial Cell Line-Derived Neurotrophic Factor; C526007:NODAL protein, human; D055457:Nodal Protein; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D019938:Cyclin D1; C498932:ACVR1B protein, human; C424205:ACVR1C protein, human; C579679:ACVR2B protein, human; D030201:Activin Receptors, Type I; D030301:Activin Receptors, Type II