Alanine and arginine rich domain containing protein, Aard, is directly regulated by androgen receptor in mouse Sertoli cells

Geng,Qiang; Ni,Li‑Wei; Ouyang,Bin; Hu,Yan‑Hua; Zhao,Yu; Guo,Jun

doi:10.3892/mmr.2016.6028

Alanine and arginine rich domain containing protein, Aard, is directly regulated by androgen receptor in mouse Sertoli cells

Authors:
- Qiang Geng
- Li‑Wei Ni
- Bin Ouyang
- Yan‑Hua Hu
- Yu Zhao
- Jun Guo
View Affiliations

Affiliations: First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300193, P.R. China, Union Stem Cell & Gene Engineering Co., Ltd, Tianjin 300384, P.R. China, Graduate School of Tianjin University of Traditional Chinese Medicine, Tianjin 300193, P.R. China, Department of Andrology, Xiyuan Hospital of China Academy of Chinese Medical Science, Beijing 100091, P.R. China
Published online on: December 12, 2016 https://doi.org/10.3892/mmr.2016.6028
Pages: 352-358

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Alanine and arginine rich domain containing protein (Aard) is specifically expressed in Sertoli cells (SCs) of mouse testis and the expression increases in an age‑dependent manner. A number of previous studies have indicated that androgen and androgen receptor (AR) signaling pathways are particularly important for spermatogenesis in mouse SCs, however, the association between Aard and AR remain to be elucidated. The present study identified Aard as a gene that is directly regulated by AR in mouse SCs, which is important in spermatogenesis. The expression of AARD was significantly downregulated in the testes of Sertoli cell‑selective AR knockout mice compared with wild‑type mice as analyzed by western blotting and immunofluorescence analyses. Quantitative polymerase chain reaction and western blotting indicated that AARD was predominantly expressed in adult mouse testis and its expression was increased in an age-dependent manner. In addition, AARD expression was upregulated by testosterone in primary SCs in vitro, which was confirmed by bioinformatics analysis and a dual‑luciferase reporter assay. Finally, chromatin immunoprecipitation and electrophoretic mobility shift assays indicated that the ligand‑bound AR activated Aard transcription via directly binding to the androgen‑responsive element of the Aard promoter. To the best of our knowledge, the present study is the first to document that Aard is directly regulated by AR in mouse Sertoli cells.

View Figures

View References

1	Heemers HV and Tindall DJ: Androgen receptor (AR) coregulators: A diversity of functions converging on and regulating the AR transcriptional complex. Endocr Rev. 28:778–808. 2007. View Article : Google Scholar : PubMed/NCBI
2	Patrão MT, Silva EJ and Avellar MC: Androgens and the male reproductive tract: An overview of classical roles and current perspectives. Arq Bras Endocrinol Metabol. 53:934–945. 2009.PubMed/NCBI
3	Lazaros L, Xita N, Takenaka A, Sofikitis N, Makrydimas G, Stefos T, Kosmas I, Zikopoulos K, Hatzi E and Georgiou I: Semen quality is influenced by androgen receptor and aromatase gene synergism. Hum Reprod. 27:3385–3392. 2012. View Article : Google Scholar : PubMed/NCBI
4	Heinlein CA and Chang C: Androgen receptor (AR) coregulators: An overview. Endocr Rev. 23:175–200. 2002. View Article : Google Scholar : PubMed/NCBI
5	Nantermet PV, Xu J, Yu Y, Hodor P, Holder D, Adamski S, Gentile MA, Kimmel DB, Harada S, Gerhold D, et al: Identification of genetic pathways activated by the androgen receptor during the induction of proliferation in the ventral prostate gland. J Biol Chem. 279:1310–1322. 2004. View Article : Google Scholar : PubMed/NCBI
6	Marcelli M, Ittmann M, Mariani S, Sutherland R, Nigam R, Murthy L, Zhao Y, DiConcini D, Puxeddu E, Esen A, et al: Androgen receptor mutations in prostate cancer. Cancer Res. 60:944–949. 2000.PubMed/NCBI
7	Chen MJ, Vu BM, Axelrad M, Dietrich JE, Gargollo P, Gunn S, Macias CG, McCullough LB, Roth DR, Sutton VR and Karaviti LP: Androgen Insensitivity Syndrome: Management Considerations from Infancy to Adulthood. Pediatr Endocrinol Rev. 12:373–387. 2015.PubMed/NCBI
8	Kim Y, Jae E and Yoon M: Influence of Androgen Receptor Expression on the Survival Outcomes in Breast Cancer: A Meta-Analysis. J Breast Cancer. 18:134–142. 2015. View Article : Google Scholar : PubMed/NCBI
9	Yeh S, Tsai MY, Xu Q, Mu XM, Lardy H, Huang KE, Lin H, Yeh SD, Altuwaijri S, Zhou X, et al: Generation and characterization of androgen receptor knockout (ARKO) mice: An in vivo model for the study of androgen functions in selective tissues. Proc Natl Acad Sci USA. 99:13498–13503. 2002. View Article : Google Scholar : PubMed/NCBI
10	De Gendt K, Swinnen JV, Saunders PT, Schoonjans L, Dewerchin M, Devos A, Tan K, Atanassova N, Claessens F, Lécureuil C, et al: A Sertoli cell-selective knockout of the androgen receptor causes spermatogenic arrest in meiosis. Proc Natl Acad Sci USA. 101:1327–1332. 2004. View Article : Google Scholar : PubMed/NCBI
11	Wang RS, Yeh S, Tzeng CR and Chang C: Androgen receptor roles in spermatogenesis and fertility: Lessons from testicular cell-specific androgen receptor knockout mice. Endocr Rev. 30:119–132. 2009. View Article : Google Scholar : PubMed/NCBI
12	Chang C, Lee SO, Wang RS, Yeh S and Chang TM: Androgen receptor (AR) physiological roles in male and female reproductive systems: Lessons learned from AR-knockout mice lacking AR in selective cells. Biol Reprod. 89:212013. View Article : Google Scholar : PubMed/NCBI
13	Mou L, Zhang Q, Wang Y, Zhang Q, Sun L, Li C, Huang W, Yuan Y, Duan Y, Diao R, et al: Identification of Ube2b as a novel target of androgen receptor in mouse sertoli cells. Biol Reprod. 89:322013. View Article : Google Scholar : PubMed/NCBI
14	Yang L, Wang Y, Zhang Q, Lai Y, Li C, Zhang Q, Huang W, Duan Y, Jiang Z, Li X, et al: Identification of Hsf1 as a novel androgen receptor-regulated gene in mouse Sertoli cells. Mol Reprod Dev. 81:514–523. 2014. View Article : Google Scholar : PubMed/NCBI
15	Bhardwaj A, Rao MK, Kaur R, Buttigieg MR and Wilkinson MF: GATA factors and androgen receptor collaborate to transcriptionally activate the Rhox5 homeobox gene in Sertoli cells. Mol Cell Biol. 28:2138–2153. 2008. View Article : Google Scholar : PubMed/NCBI
16	De Gendt K, Denolet E, Willems A, Daniels VW, Clinckemalie L, Denayer S, Wilkinson MF, Claessens F, Swinnen JV and Verhoeven G: Expression of Tubb3, a beta-tubulin isotype, is regulated by androgens in mouse and rat Sertoli cells. Biol Reprod. 85:934–945. 2011. View Article : Google Scholar : PubMed/NCBI
17	Svingen T, Beverdam A, Verma P, Wilhelm D and Koopman P: Aard is specifically up-regulated in Sertoli cells during mouse testis differentiation. Int J Dev Biol. 51:255–258. 2007. View Article : Google Scholar : PubMed/NCBI
18	Zhang QX, Zhang XY, Zhang ZM, Lu W, Liu L, Li G, Cai ZM, Gui YT and Chang C: Identification of testosterone-/androgen receptor-regulated genes in mouse Sertoli cells. Asian J Androl. 14:294–300. 2012. View Article : Google Scholar : PubMed/NCBI
19	Verhoeven G and Cailleau J: Testicular peritubular cells secrete a protein under androgen control that inhibits induction of aromatase activity in Sertoli cells. Endocrinology. 123:2100–2110. 1988. View Article : Google Scholar : PubMed/NCBI
20	Lutfalla G and Uze G: Performing quantitative reverse-transcribed polymerase chain reaction experiments. Methods Enzymol. 410:386–400. 2006. View Article : Google Scholar : PubMed/NCBI
21	Kim JM, Hwang SH, Song EJ, Lee SY, Kim YD, Lee CH, Lee MK, Chang CL and Lee EY: Comparative quantification of plasma hnRNP B1 mRNA in non-small cell lung cancer patients by real-time PCR. Korean J Lab Med. 29:249–255. 2009.(In Korean). View Article : Google Scholar : PubMed/NCBI
22	Fujii H and Fujita T: Isolation of Specific Genomic Regions and Identification of Their Associated Molecules by Engineered DNA-Binding Molecule-Mediated Chromatin Immunoprecipitation (enChIP) Using the CRISPR System and TAL Proteins. Int J Mol Sci. 16:21802–21812. 2015. View Article : Google Scholar : PubMed/NCBI
23	Smith LB and Walker WH: The regulation of spermatogenesis by androgens. Semin Cell Dev Biol. 30:2–13. 2014. View Article : Google Scholar : PubMed/NCBI
24	Walker WH: Testosterone signaling and the regulation of spermatogenesis. Spermatogenesis. 1:116–120. 2011. View Article : Google Scholar : PubMed/NCBI
25	Blomberg LA, Chan WY, Clerch LB and Massaro D: Molecular cloning and characterization of a novel gene upregulated early during postnatal rat lung development. Biochim Biophys Acta. 1574:391–398. 2002. View Article : Google Scholar : PubMed/NCBI
26	Wang Q, Li W, Liu XS, Carroll JS, Jänne OA, Keeton EK, Chinnaiyan AM, Pienta KJ and Brown M: A hierarchical network of transcription factors governs androgen receptor-dependent prostate cancer growth. Mol Cell. 27:380–392. 2007. View Article : Google Scholar : PubMed/NCBI
27	Spillane M, Schwarz N and Willoughby DS: Upper-body resistance exercise augments vastus lateralis androgen receptor-DNA binding and canonical Wnt/β-catenin signaling compared to lower-body resistance exercise in resistance-trained men without an acute increase in serum testosterone. Steroids. 98:63–71. 2015. View Article : Google Scholar : PubMed/NCBI