A potential role for zinc transporter 7 in testosterone synthesis in mouse Leydig tumor cells

Chu,Qingqing; Chi,Zhi-Hong; Zhang,Xiuli; Liang,Dan; Wang,Xuemei; Zhao,Yue; Zhang,Li; Zhang,Ping

doi:10.3892/ijmm.2016.2576

A potential role for zinc transporter 7 in testosterone synthesis in mouse Leydig tumor cells

Authors:
- Qingqing Chu
- Zhi-Hong Chi
- Xiuli Zhang
- Dan Liang
- Xuemei Wang
- Yue Zhao
- Li Zhang
- Ping Zhang
View Affiliations

Affiliations: Department of Pathophysiology, China Medical University, Shenyang, Liaoning, P.R. China, Department of Nephrology, Benxi Center Hospital, China Medical University, Benxi, Liaoning, P.R. China, Troops of 95935 Unit, Harbin, Heilongjiang, P.R. China, School of Stomatology of China Medical University, Shenyang, Liaoning, P.R. China, Department of Histology and Embryology, Liaoning Medical College, Jinzhou, Liaoning, P.R. China
Published online on: April 25, 2016 https://doi.org/10.3892/ijmm.2016.2576
Pages: 1619-1626

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

Previous studies have demonstrated that zinc (Zn) is an essential trace element which is involved in male reproduction. The zinc transporter (ZnT) family, SLC30a, is involved in the maintenance of Zn homeostasis and in mediating intracellular signaling events; however, relatively little is known regarding the effect of ZnTs on testosterone synthesis. Thus, in the present study, we aimed to determine the effect of Zn transporter 7 (ZnT7) on testosterone synthesis in male CD-1 mice and mouse Leydig cells. The findings of the present study revealed that the concentrations of Zn in the testes and Leydig cells were significantly lower in mice fed a Zn-deficient diet compared with the control mice fed a Zn-adequate diet. In addition, ZnT7 was principally expressed and colocalized with steroidogenic acute regulatory protein (StAR) in the Leydig cells of male CD-1 mice. ZnT7 expression was downregulated in the mice fed a Zn-deficient diet, which led to decreases in the expression of the enzymes involved in testosterone synthesis namely cholesterol side‑chain cleavage enzyme (P450scc) and 3β-hydroxysteroid dehydrogenase/D5-D4 isomerase (3β-HSD) as well as decreased serum testosterone levels. These results suggested that Znt7 may be involved in testosterone synthesis in the mouse testes. To examine this hypothesis, we used the mouse Leydig tumor cell line (MLTC-1 cell line) in which the ZnT7 gene had been silenced, in order to gauge the impact of changes in ZnT7 expression on testosterone secretion and the enzymes involved in testosterone synthesis. The results demonstrated that ZnT7 gene silencing downregulated the expression of StAR, P450scc and 3β-HSD as well as progesterone concentrations in the human chorionic gonadotrophin (hCG)-stimulated MLTC-1 cells. Taken together, these findings reveal that ZnT7 may play an important role in the regulation of testosterone synthesis by modulating steroidogenic enzymes, and may represent a therapeutic target in testosterone deficiency.

View Figures

View References

1	Hidiroglou M and Knipfel JE: Zinc in mammalian sperm: a review. J Dairy Sci. 67:1147–1156. 1984. View Article : Google Scholar : PubMed/NCBI
2	Coleman JE: Zinc proteins: enzymes, storage proteins, transcription factors, and replication proteins. Annu Rev Biochem. 61:897–946. 1992. View Article : Google Scholar : PubMed/NCBI
3	Hartoma TR, Nahoul K and Netter A: Zinc, plasma androgens and male sterility. Lancet. 2:1125–1126. 1977. View Article : Google Scholar : PubMed/NCBI
4	Aihara K, Nishi Y, Hatano S, Kihara M, Ohta M, Sakoda K, Uozumi T and Usui T: Zinc, copper, manganese, and selenium metabolism in patients with human growth hormone deficiency or acromegaly. J Pediatr Gastroenterol Nutr. 4:610–615. 1985. View Article : Google Scholar : PubMed/NCBI
5	Bedwal RS and Bahuguna A: Zinc, copper and selenium in reproduction. Experientia. 50:626–640. 1994. View Article : Google Scholar : PubMed/NCBI
6	El Hendy HA, Yousef MI and Abo El-Naga NI: Effect of dietary zinc deficiency on hematological and biochemical parameters and concentrations of zinc, copper, and iron in growing rats. Toxicology. 167:163–170. 2001. View Article : Google Scholar : PubMed/NCBI
7	Matossian MK: Fertility decline in Europe 1875–1913. Was zinc deficiency the cause? Perspect Biol Med. 34:604–616. 1991. View Article : Google Scholar : PubMed/NCBI
8	Brown KH and Wuehler SE: Zinc and human health: results of recent trials and implications for program interventions and research. Micronutrient Initiative; Ottawa: 2000
9	Hesketh JE: Effects of dietary zinc deficiency on Leydig cell ultrastructure in the boar. J Comp Pathol. 92:239–247. 1982. View Article : Google Scholar : PubMed/NCBI
10	Lei KY, Abbasi A and Prasad AS: Function of pituitary-gonadal axis in zinc-deficient rats. Am J Physiol. 230:1730–1732. 1976.PubMed/NCBI
11	Hadwan MH, Almashhedy LA and Alsalman AR: Study of the effects of oral zinc supplementation on peroxynitrite levels, arginase activity and NO synthase activity in seminal plasma of Iraqi asthenospermic patients. Reprod Biol Endocrinol. 12:12014. View Article : Google Scholar : PubMed/NCBI
12	Abbasi AA, Prasad AS and Rabbani PR: Experimental zinc deficiency in man: effect on spermatogenesis. Trans Assoc Am Physicians. 92:292–302. 1979.PubMed/NCBI
13	Khan MS, Zaman S, Sajjad M, Shoaib M and Gilani G: Assessment of the level of trace element zinc in seminal plasma of males and evaluation of its role in male infertility. Int J Appl Basic Med Res. 1:93–96. 2011. View Article : Google Scholar : PubMed/NCBI
14	Chasapis CT, Loutsidou AC, Spiliopoulou CA and Stefanidou ME: Zinc and human health: an update. Arch Toxicol. 86:521–534. 2012. View Article : Google Scholar
15	Kambe T, Yamaguchi-Iwai Y, Sasaki R and Nagao M: Overview of mammalian zinc transporters. Cell Mol Life Sci. 61:49–68. 2004. View Article : Google Scholar : PubMed/NCBI
16	Palmiter RD and Huang L: Efflux and compartmentalization of zinc by members of the SLC30 family of solute carriers. Pflugers Arch. 447:744–751. 2004. View Article : Google Scholar
17	Kambe T, Hashimoto A and Fujimoto S: Current understanding of ZIP and ZnT zinc transporters in human health and diseases. Cell Mol Life Sci. 71:3281–3295. 2014. View Article : Google Scholar : PubMed/NCBI
18	Palmiter RD, Cole TB, Quaife CJ and Findley SD: ZnT-3, a putative transporter of zinc into synaptic vesicles. Proc Natl Acad Sci USA. 93:14934–14939. 1996. View Article : Google Scholar : PubMed/NCBI
19	Cole TB, Wenzel HJ, Kafer KE, Schwartzkroin PA and Palmiter RD: Elimination of zinc from synaptic vesicles in the intact mouse brain by disruption of the ZnT3 gene. Proc Natl Acad Sci USA. 96:1716–1721. 1999. View Article : Google Scholar : PubMed/NCBI
20	Kambe T, Narita H, Yamaguchi-Iwai Y, Hirose J, Amano T, Sugiura N, Sasaki R, Mori K, Iwanaga T and Nagao M: Cloning and characterization of a novel mammalian zinc transporter, zinc transporter 5, abundantly expressed in pancreatic beta cells. J Biol Chem. 277:19049–19055. 2002. View Article : Google Scholar : PubMed/NCBI
21	Kirschke CP and Huang L: ZnT7, a novel mammalian zinc transporter, accumulates zinc in the Golgi apparatus. J Biol Chem. 278:4096–4102. 2003. View Article : Google Scholar
22	Lemaire K, Chimienti F and Schuit F: Zinc transporters and their role in the pancreatic β-cell. J Diabetes Investig. 3:202–211. 2012. View Article : Google Scholar : PubMed/NCBI
23	Chimienti F, Devergnas S, Favier A and Seve M: Identification and cloning of a beta-cell-specific zinc transporter, ZnT-8, localized into insulin secretory granules. Diabetes. 53:2330–2337. 2004. View Article : Google Scholar : PubMed/NCBI
24	Kambe T, Weaver BP and Andrews GK: The genetics of essential metal homeostasis during development. Genesis. 46:214–228. 2008. View Article : Google Scholar : PubMed/NCBI
25	Jiang MH, Cai B, Tuo Y, Wang J, Zang ZJ, Tu X, Gao Y, Su Z, Li W, Li G, et al: Characterization of Nestin-positive stem Leydig cells as a potential source for the treatment of testicular Leydig cell dysfunction. Cell Res. 24:1466–1485. 2014. View Article : Google Scholar : PubMed/NCBI
26	Wang H, Wang Q, Zhao XF, Liu P, Meng XH, Yu T, Ji YL, Zhang H, Zhang C, Zhang Y and Xu DX: Cypermethrin exposure during puberty disrupts testosterone synthesis via downregu-lating StAR in mouse testes. Arch Toxicol. 84:53–61. 2010. View Article : Google Scholar
27	Caron KM, Soo SC, Wetsel WC, Stocco DM, Clark BJ and Parker KL: Targeted disruption of the mouse gene encoding steroidogenic acute regulatory protein provides insights into congenital lipoid adrenal hyperplasia. Proc Natl Acad Sci USA. 94:11540–11545. 1997. View Article : Google Scholar : PubMed/NCBI
28	King SR, Manna PR, Ishii T, Syapin PJ, Ginsberg SD, Wilson K, Walsh LP, Parker KL, Stocco DM, Smith RG and Lamb DJ: An essential component in steroid synthesis, the steroidogenic acute regulatory protein, is expressed in discrete regions of the brain. J Neurosci. 22:10613–10620. 2002.PubMed/NCBI
29	Stocco DM and Clark BJ: The role of the steroidogenic acute regulatory protein in steroidogenesis. Steroids. 62:29–36. 1997. View Article : Google Scholar : PubMed/NCBI
30	Raucci F, D'Aniello A and Di Fiore MM: Stimulation of androgen production by D-aspartate through the enhancement of StAR, P450scc and 3β-HSD mRNA levels in vivo rat testis and in culture of immature rat Leydig cells. Steroids. 84:103–110. 2014. View Article : Google Scholar : PubMed/NCBI
31	Stocco DM: StAR protein and the regulation of steroid hormone biosynthesis. Annu Rev Physiol. 63:193–213. 2001. View Article : Google Scholar : PubMed/NCBI
32	Hu Y, Dong C, Chen M, Lu J, Han X, Qiu L, Chen Y, Qin J, Li X, Gu A, et al: Low-dose monobutyl phthalate stimulates steroidogenesis through steroidogenic acute regulatory protein regulated by SF-1, GATA-4 and C/EBP-beta in mouse Leydig tumor cells. Reprod Biol Endocrinol. 11:722013. View Article : Google Scholar : PubMed/NCBI
33	Pakarinen P, Vihko KK, Voutilainen R and Huhtaniemi I: Differential response of luteinizing hormone receptor and steroidogenic enzyme gene expression to human chorionic gonadotropin stimulation in the neonatal and adult rat testis. Endocrinology. 127:2469–2474. 1990. View Article : Google Scholar : PubMed/NCBI
34	Baba Y: Effect of HCG administration in the gossypol-treated male rats. Hinyokika Kiyo. 34:643–647. 1988.In Japanese. PubMed/NCBI
35	Scott IS, Charlton HM, Cox BS, Grocock CA, Sheffield JW and O'Shaughnessy PJ: Effect of LH injections on testicular steroidogenesis, cholesterol side-chain cleavage P450 mRNA content and Leydig cell morphology in hypogonadal mice. J Endocrinol. 125:131–138. 1990. View Article : Google Scholar : PubMed/NCBI
36	Xu Z, Kawai M, Bandiera SM and Chang TK: Influence of dietary zinc deficiency during development on hepatic CYP2C11, CYP2C12, CYP3A2, CYP3A9, and CYP3A18 expression in postpubertal male rats. Biochem Pharmacol. 62:1283–1291. 2001. View Article : Google Scholar : PubMed/NCBI
37	Croxford TP, McCormick NH and Kelleher SL: Moderate zinc deficiency reduces testicular Zip6 and Zip10 abundance and impairs spermatogenesis in mice. J Nutr. 141:359–365. 2011. View Article : Google Scholar : PubMed/NCBI
38	Favier AE: The role of zinc in reproduction. Hormonal mechanisms. Biol Trace Elem Res. 32:363–382. 1992. View Article : Google Scholar : PubMed/NCBI
39	Om AS and Chung KW: Dietary zinc deficiency alters 5 alpha-reduction and aromatization of testosterone and androgen and estrogen receptors in rat liver. J Nutr. 126:842–848. 1996.PubMed/NCBI
40	Qureshi IZ and Abbas Q: Modulation of testicular and whole blood trace element concentrations in conjunction with testosterone release following kisspeptin administration in male rabbits (Oryctolagus cuniculus). Biol Trace Elem Res. 154:210–216. 2013. View Article : Google Scholar : PubMed/NCBI
41	Sunderman FW Jr: The influence of zinc on apoptosis. Ann Clin Lab Sci. 25:134–142. 1995.PubMed/NCBI
42	Vallee BL and Falchuk KH: The biochemical basis of zinc physiology. Physiol Rev. 73:79–118. 1993.PubMed/NCBI
43	Prasad AS: Clinical and biochemical manifestation zinc deficiency in human subjects. J Pharmacol. 16:344–352. 1985.PubMed/NCBI
44	Root AW, Duckett G, Sweetland M and Reiter EO: Effects of zinc deficiency upon pituitary function in sexually mature and immature male rats. J Nutr. 109:958–964. 1979.PubMed/NCBI
45	Huang L and Tepaamorndech S: The SLC30 family of zinc transporters - a review of current understanding of their biological and pathophysiological roles. Mol Aspects Med. 34:548–560. 2013. View Article : Google Scholar : PubMed/NCBI
46	Kelleher SL, Seo YA and Lopez V: Mammary gland zinc metabolism: regulation and dysregulation. Genes Nutr. 4:83–94. 2009. View Article : Google Scholar : PubMed/NCBI
47	Chi ZH, Feng WY, Gao HL, Zheng W, Huang L and Wang ZY: ZNT7 and Zn²⁺ are present in different cell populations in the mouse testis. Histol Histopathol. 24:25–30. 2009.
48	Chi ZH, Ren H, Wang X, Rong M, Huang L and Wang ZY: The cellular and subcellular localization of zinc transporter 7 in the mouse spinal cord. Histol Histopathol. 23:781–787. 2008.PubMed/NCBI
49	Huang L, Yan M and Kirschke CP: Over-expression of ZnT7 increases insulin synthesis and secretion in pancreatic beta-cells by promoting insulin gene transcription. Exp Cell Res. 316:2630–2643. 2010. View Article : Google Scholar : PubMed/NCBI
50	Mukherjee S, Chiu R, Leung SM and Shields D: Fragmentation of the Golgi apparatus: an early apoptotic event independent of the cytoskeleton. Traffic. 8:369–378. 2007. View Article : Google Scholar : PubMed/NCBI
51	Chi ZH, Wang X, Wang ZY, Gao HL, Dahlstrom A and Huang L: Zinc transporter 7 is located in the cis-Golgi apparatus of mouse choroid epithelial cells. Neuroreport. 17:1807–1811. 2006. View Article : Google Scholar : PubMed/NCBI