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Abstract

Volume 17, Issue 6 (November 2015) 17, 996–1005; 10.4103/1008-682X.159722

NODAL secreted by male germ cells regulates the proliferation and function of human Sertoli cells from obstructive azoospermia and nonobstructive azoospermia patients

Ru-Hui Tian, Shi Yang, Zi-Jue Zhu, Jun-Long Wang, Yun Liu, Chencheng Yao, Meng Ma, Ying Guo, Qingqing Yuan, Yanan Hai, Yi-Ran Huang, Zuping He, Zheng Li

Department of Urology, Institute of Andrology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China
2 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
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

Correspondence: Dr. Z Li

19-Aug-2015

Abstract

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.

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