CC BY-NC-ND 4.0 · Ann Natl Acad Med Sci 2020; 56(01): 30-37
DOI: 10.1055/s-0040-1709091
Original Article

A Novel Antigonadotropic Role of Thyroid Stimulating Hormone on Leydig Cell-Derived Mouse Leydig Tumor Cells-1 Line

Bodhana Dhole
1   Department of Reproductive Biology, All India Institute of Medical Sciences, New Delhi, India
,
Surabhi Gupta
1   Department of Reproductive Biology, All India Institute of Medical Sciences, New Delhi, India
,
Skand Shekhar
2   Section on Endocrinology and Genetics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, United States
,
Anand Kumar*
1   Department of Reproductive Biology, All India Institute of Medical Sciences, New Delhi, India
› Author Affiliations
Funding None.

Abstract

Subclinical hypothyroid men characterized by a rise in only thyroid stimulating hormone (TSH) levels, and normal thyroid hormone levels showed a fall in their serum progesterone and testosterone levels. This suggested a role of TSH in regulating Leydig cell steroidogenesis. Therefore, we investigated the direct role of TSH on steroid production and secretion using a mouse Leydig tumor cell line-1 (MLTC-1). MLTC-1 cells were treated with different doses of TSH isolated from porcine pituitary as well as recombinant TSH. Steroid secretion was measured by radioimmunoassay (RIA). The mRNA levels of steroidogenic enzymes were quantitated by real-time polymerase chain reaction (RT-PCR), whereas the corresponding protein levels were determined by western blot. In MLTC-1 cells, pituitary TSH as well as recombinant TSH inhibited progesterone and testosterone secretion in a dose-dependent manner. The inhibitory action of TSH on steroid secretion was unique and not mimicked by other anterior pituitary hormones including follicle stimulating hormone and adrenocorticotropic hormone. Recombinant TSH showed no effect on steroidogenic acute regulatory protein and CYP11A1, the enzymes catalyzing the nonsteroidogenic and steroidogenic rate-limiting steps of steroid synthesis, respectively. Recombinant TSH was shown to inhibit steroidogenesis in MLTC-1 cells by inhibiting the 3-β hydroxy steroid dehydrogenase mRNA and protein levels, the enzyme that catalyzes the conversion of pregnenolone to progesterone. This inhibitory effect of TSH is probably direct as both mRNA and protein of the TSH receptor were shown to be present in the MLTC-1 cells.

* Retired




Publication History

Article published online:
20 April 2020

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  • References

  • 1 Manna PR, Kero J, Tena-Sempere M, Pakarinen P, Stocco DM, Huhtaniemi IT. Assessment of mechanisms of thyroid hormone action in mouse Leydig cells: regulation of the steroidogenic acute regulatory protein, steroidogenesis, and luteinizing hormone receptor function. Endocrinology 2001; 142 (01) 319-331
  • 2 Manna PR, Tena-Sempere M, Huhtaniemi IT. Molecular mechanisms of thyroid hormone-stimulated steroidogenesis in mouse leydig tumor cells. Involvement of the steroidogenic acute regulatory (StAR) protein. J Biol Chem 1999; 274 (09) 5909-5918
  • 3 Maran RR, Arunakaran J, Aruldhas MM. T3 directly stimulates basal and modulates LH induced testosterone and oestradiol production by rat Leydig cells in vitro. Endocr J 2000; 47 (04) 417-428
  • 4 Kumar A, Mohanty BP, Rani L. Secretion of testicular steroids and gonadotrophins in hypothyroidism. Andrologia 2007; 39 (06) 253-260
  • 5 Teerds KJ, de Rooij DG, de Jong FH, van Haaster LH. Development of the adult-type Leydig cell population in the rat is affected by neonatal thyroid hormone levels. Biol Reprod 1998; 59 (02) 344-350
  • 6 Hardy MP, Sharma RS, Arambepola NK. et al. Increased proliferation of Leydig cells induced by neonatal hypothyroidism in the rat. J Androl 1996; 17 (03) 231-238
  • 7 Cristovão FC, Bisi H, Mendonça BB, Bianco AC, Bloise W. Severe and mild neonatal hypothyroidism mediate opposite effects on Leydig cells of rats. Thyroid 2002; 12 (01) 13-18
  • 8 Rao JN, Liang JY, Chakraborti P, Feng P. Effect of thyroid hormone on the development and gene expression of hormone receptors in rat testes in vivo. J Endocrinol Invest 2003; 26 (05) 435-443
  • 9 Kumar A, Chaturvedi PK, Mohanty BP. Hypoandrogenaemia is associated with subclinical hypothyroidism in men. Int J Androl 2007; 30 (01) 14-20
  • 10 Mendis-Handagama SM, Siril Ariyaratne HB. Leydig cells, thyroid hormones and steroidogenesis. Indian J Exp Biol 2005; 43 (11) 939-962
  • 11 Rebois RV. Establishment of gonadotropin-responsive murine leydig tumor cell line. J Cell Biol 1982; 94 (01) 70-76
  • 12 Cascio C, Prasad VV, Lin YY, Lieberman S, Papadopoulos V. Detection of P450c17-independent pathways for dehydroepiandrosterone (DHEA) biosynthesis in brain glial tumor cells. Proc Natl Acad Sci U S A 1998; 95 (06) 2862-2867
  • 13 O’Shaughnessy PJ, Morris ID, Baker PJ. Leydig cell re-generation and expression of cell signaling molecules in the germ cell-free testis. Reproduction 2008; 135 (06) 851-858
  • 14 Marians RC, Ng L, Blair HC, Unger P, Graves PN, Davies TF. Defining thyrotropin-dependent and -independent steps of thyroid hormone synthesis by using thyrotropin receptor-null mice. Proc Natl Acad Sci U S A 2002; 99 (24) 15776-15781
  • 15 Konieczna A, Szczepańska A, Sawiuk K, Węgrzyn G, Łyżeń R. Effects of partial silencing of genes coding for enzymes involved in glycolysis and tricarboxylic acid cycle on the enterance of human fibroblasts to the S phase. BMC Cell Biol 2015; 16: 16
  • 16 Mehendale RG, Bruot BC. Thyroid stimulating hormone inhibits rat granulosa cell steroidogenesis in primary culture. Endocrine 1995; 3 (03) 215-220
  • 17 Agard JA, Duffy DM, Jacot T, Archer DF. Thyroid stimulating hormone (TSH) receptor on granulosa cells. Fertil Steril 2011; 96 (03) S118
  • 18 Clark BJ, Wells J, King SR, Stocco DM. The purification, cloning, and expression of a novel luteinizing hormone-induced mitochondrial protein in MA-10 mouse Leydig tumor cells. Characterization of the steroidogenic acute regulatory protein (StAR). J Biol Chem 1994; 269 (45) 28314-28322
  • 19 Meinsohn MC, Smith OE, Bertolin K, Murphy BD. The orphan nuclear receptors steroidogenic factor-1 and liver receptor homolog-1: structure, regulation, and essential roles in mammalian reproduction. Physiol Rev 2019; 99 (02) 1249-1279
  • 20 Zirkin BR, Papadopoulos V. Leydig cells: formation, function, and regulation. Biol Reprod 2018; 99 (01) 101-111
  • 21 Hu J, Zhang Z, Shen WJ, Azhar S. Cellular cholesterol delivery, intracellular processing and utilization for biosynthesis of steroid hormones. Nutr Metab (Lond) 2010; 7: 47
  • 22 Acconcia F, Marino M. Steroid hormones: synthesis, secretion, and transport. In: Belfiore A, LeRoith D. eds. Principles of Endocrinology and Hormone Action. Cham: Springer International Publishing; 2016: 1-31
  • 23 Hall PF. Testicular steroid synthesis: organization and regulation. In: Knobil E, Neill J. eds. The Physiology of Reproduction. 2nd ed. New York, NY: Raven Press; 1994: 1335-1362
  • 24 Payne AH, Abbaszade IG, Clarke TR, Bain PA, Park CH. The multiple murine 3 beta-hydroxysteroid dehydrogenase isoforms: structure, function, and tissue- and developmentally specific expression. Steroids 1997; 62 (01) 169-175
  • 25 Fadlalla MB, Wei Q, Fedail JS, Mehfooz A, Mao D, Shi F. Effects of hyper- and hypothyroidism on the development and proliferation of testicular cells in prepubertal rats. Anim Sci J 2017; 88 (12) 1943-1954
  • 26 Shekhar SDB, Gupta S, Kumar A. A unique anti-gonadotropic effect of TSH on Leydig cell derived Mltc-1 Line.The Endocrine Society Meeting. Available at: https://endo.confex.com/endo/2017endo/meetingapp.cgi/Paper/31402. Accessed April 3, 2017
  • 27 Sun SC, Hsu PJ, Wu FJ, Li SH, Lu CH, Luo CW. Thyrostimulin, but not thyroid-stimulating hormone (TSH), acts as a paracrine regulator to activate the TSH receptor in mammalian ovary. J Biol Chem 2010; 285 (06) 3758-3765
  • 28 Allgeier A, Offermanns S, Van J Sande, Spicher K, Schultz G, Dumont JE. The human thyrotropin receptor activates G-proteins Gs and Gq/11. J Biol Chem 1994; 269 (19) 13733-13735
  • 29 Krude H, Biebermann H. The thyroid and its regulation by the TSHR: evolution, development, and congenital defects. In: Luster M, Duntas LH, Wartofsky L. eds. The Thyroid and Its Diseases: A Comprehensive Guide for the Clinician. Cham: Springer International Publishing; 2019: 219-233
  • 30 Reyland ME. Protein kinase C is a tonic negative regulator of steroidogenesis and steroid hydroxylase gene expression in Y1 adrenal cells and functions independently of protein kinase A. Mol Endocrinol 1993; 7 (08) 1021-1030
  • 31 LeHoux JG, Dupuis G, Lefebvre A. Control of CYP11B2 gene expression through differential regulation of its promoter by atypical and conventional protein kinase C isoforms. J Biol Chem 2001; 276 (11) 8021-8028