Mitochondrial sirtuins in the rat adrenal gland: location within the glands of males and females, hormonal and developmental regulation of gene expressions
Abstract
Introduction. Sirtuins are NAD dependent class III histone deacetylases. In adrenal cortex mitochondria are able to transform — via nicotinamide nucleotide transhydrogenase (NNT) — NAD into NADPH, which is required for steroidogenesis. These findings suggest that sirtuins expressed in mitochondria, Sirt3, Sirt4 and Sirt5, may be associated with adrenal steroidogenesis. Therefore, the purpose of this study was to characterize the expression of mitochondrial sirtuins (Sirt3–5) in individual compartments of rat adrenal cortex, their developmental regulation and to demonstrate whether their expression is dependent on adrenocorticotrophic hormone (ACTH) and Nampt (nicotinamide phosphoribosyltransferase also known as visfatin/PBEF), the rate-limiting enzyme in the regulation of mammalian NAD synthesis.
Material and methods. Studies were performed on rat adrenal glands or on primary culture of rat adrenocortical cells. Expression of mitochondrial sirtuins (Sirt3–5) was evaluated by Affymetrix microarray system or QPCR. The bulk of data were extracted from our earlier experiments which have been reanalyzed in regard to Sirt3–5 mRNAs expression levels and — if necessary — validated by QPCR.
Results. Sirt3–5 were expressed throughout the rat adrenal, with the highest expression level of Sirt5. The level of expression of all sirtuins is higher in the zona glomerulosa (ZG) and zona fasciculata/reticularis (ZF/R) than in the adrenal medulla. Sirt3 and Sirt5 expression levels were similar in adult male and female rats, while Sirt4 expression level was higher in females. As revealed by analysis of the available open database, no significant changes in Sirt3–5 expression levels in whole adrenal glands were observed up to week 104 of life of both male and female rats. Moreover, 60 min after intraperitoneal ACTH injection the expression level of Sirt3 in the entire gland was elevated while Sirt5 expression level lowered. On the other hand, chronic ACTH infusion (48 h) did not change expression of studied sirtuins. In cultured cells, ACTH greatly increased the expression levels of the Sirt4 and Sirt5. In cultured cells, Fk866 — a highly specific competitive inhibitor of Nampt — reduced expression level of Sirt5 only. In enucleation-induced regenerating rat adrenal, the expression levels of all studied sirtuins were significantly reduced in relation to the control group. Finally, in primary rat adrenal culture the FCS depletion elevates the Sirt3 and Sirt4 expression levels and downregulates Sirt5 expression.
Conclusions. Sirt3–5 are expressed throughout the rat adrenal, with the highest expression levels in adrenal cortex. Performed experiments (ACTH stimulation, FCS depletion, regeneration) suggest that in the adrenal cortex, the mitochondrial Sirt5 is the primary mitochondrial sirtuin involved in regulating the biological activity of adrenocortical cells. Our results also suggest that normal levels of intracellular Nampt (iNampt) enzymatic activity are required to maintain normal (control) levels of Sirt5 mRNA in cultured cells.
Keywords: ratadrenal glandmitochondrial sirtuinsACTHontogenesisadrenal regenerationNamptmicroarraysQPCR
References
- Chang HC, Guarente L. SIRT1 and other sirtuins in metabolism. Trends Endocrinol Metab. 2014; 25(3): 138–145.
- Onyango P, Celic I, McCaffery JM, et al. SIRT3, a human SIR2 homologue, is an NAD-dependent deacetylase localized to mitochondria. Proc Natl Acad Sci USA. 2002; 99(21): 13653–13658.
- Schwer B, North BJ, Frye RA, et al. The human silent information regulator (Sir)2 homologue hSIRT3 is a mitochondrial nicotinamide adenine dinucleotide-dependent deacetylase. J Cell Biol. 2002; 158(4): 647–657.
- Michishita E, Park JY, Burneskis JM, et al. Evolutionarily conserved and nonconserved cellular localizations and functions of human SIRT proteins. Mol Biol Cell. 2005; 16(10): 4623–4635.
- Haigis MC, Guarente LP. Mammalian sirtuins — emerging roles in physiology, aging, and calorie restriction. Genes Dev. 2006; 20(21): 2913–2921.
- Schwer B, Verdin E. Conserved metabolic regulatory functions of sirtuins. Cell Metab. 2008; 7(2): 104–112.
- Satoh A, Stein L, Imai S. The role of mammalian sirtuins in the regulation of metabolism, aging, and longevity. Handb Exp Pharmacol. 2011; 206: 125–162.
- He W, Newman JC, Wang MZ, et al. Mitochondrial sirtuins: regulators of protein acylation and metabolism. Trends Endocrinol Metab. 2012; 23(9): 467–476.
- Hallows WC, Lee S, Denu JM. Sirtuins deacetylate and activate mammalian acetyl-CoA synthetases. Proc Natl Acad Sci USA. 2006; 103(27): 10230–10235.
- Osborne B, Bentley NL, Montgomery MK, et al. The role of mitochondrial sirtuins in health and disease. Free Radic Biol Med. 2016; 100: 164–174.
- Yang L, Ma X, He Y, et al. Sirtuin 5: a review of structure, known inhibitors and clues for developing new inhibitors. Sci China Life Sci. 2017; 60(3): 249–256.
- Imai SI. The NAD World: a new systemic regulatory network for metabolism and aging — Sirt1, systemic NAD biosynthesis, and their importance. Cell Biochem Biophys. 2009; 53(2): 65–74.
- Revollo JR, Grimm AA, Imai SI. The NAD biosynthesis pathway mediated by nicotinamide phosphoribosyltransferase regulates Sir2 activity in mammalian cells. J Biol Chem. 2004; 279(49): 50754–50763.
- Revollo JR, Körner A, Mills KF, et al. Nampt/PBEF/Visfatin regulates insulin secretion in beta cells as a systemic NAD biosynthetic enzyme. Cell Metab. 2007; 6(5): 363–375.
- Houtkooper RH, Cantó C, Wanders RJ, et al. The secret life of NAD+: an old metabolite controlling new metabolic signaling pathways. Endocr Rev. 2010; 31(2): 194–223.
- Artemenko IP, Jefcoate CR. Multiple contributions from long-chain fatty acid metabolism in Y-1 and MA-10 cells. Endocr Res. 2004; 30(4): 637.
- Miller WL, Auchus RJ. The molecular biology, biochemistry, and physiology of human steroidogenesis and its disorders. Endocr Rev. 2011; 32(1): 81–151.
- Miller WL. Steroid hormone synthesis in mitochondria. Mol Cell Endocrinol. 2013; 379(1-2): 62–73.
- Midzak A, Papadopoulos V. Adrenal mitochondria and steroidogenesis: from individual proteins to functional protein assemblies. Front Endocrinol (Lausanne). 2016; 7: 106.
- Payne AH, Hales DB. Overview of steroidogenic enzymes in the pathway from cholesterol to active steroid hormones. Endocr Rev. 2004; 25(6): 947–970.
- Hanukoglu I, Rapoport R. Routes and regulation of NADPH production in steroidogenic mitochondria. Endocrine Research. 2009; 21(1-2): 231–241.
- Meimaridou E., Chortis V., Fragoulis T., et al. Role of Nicotinamide nucleotide transhydrogenase in the control of steroidogenesis in mouse adrenals. Endocrine Society's 98th Annual Meeting and Expo. Boston, April 1–4, 2016.
- Weinberg-Shukron A, Abu-Libdeh A, Zhadeh F, et al. Combined mineralocorticoid and glucocorticoid deficiency is caused by a novel founder nicotinamide nucleotide transhydrogenase mutation that alters mitochondrial morphology and increases oxidative stress. J Med Genet. 2015; 52(9): 636–641.
- Roucher-Boulez F, Mallet-Motak D, Samara-Boustani D, et al. NNT mutations: a cause of primary adrenal insufficiency, oxidative stress and extra-adrenal defects. Eur J Endocrinol. 2016; 175(1): 73–84.
- Celichowski PJK, Milecka P, Szyszka M, et al. Nampt (visfatin) and hypothalamo-pituitary-adrenal (HPA) axis of the rat. Mol Med Rep (accepted to print), 2017.
- Trejter M, Jopek K, Celichowski P, et al. Expression of estrogen, estrogen related and androgen receptors in adrenal cortex of intact adult male and female rats. Folia Histochem Cytobiol. 2015; 53(2): 133–144.
- Jopek K, Celichowski P, Szyszka M, et al. Transcriptome Profile of Rat Adrenal Evoked by Gonadectomy and Testosterone or Estradiol Replacement. Front Endocrinol (Lausanne). 2017; 8: 26.
- Navolotskaya EV, Kovalitskaya YA, Zolotarev YA, et al. Binding of synthetic fragments of beta-endorphin to nonopioid beta-endorphin receptor. J Pept Sci. 2008; 14(10): 1121–1128.
- Hruby VJ, Cai M, Nyberg J, et al. Approaches to the rational design of selective melanocortin receptor antagonists. Expert Opin Drug Discov. 2011; 6(5): 543–557.
- Tyczewska M, Rucinski M, Trejter M, et al. Angiogenesis in the course of enucleation-induced adrenal regeneration — expression of selected genes and proteins involved in development of capillaries. Peptides. 2012; 38(2): 404–413.
- Tyczewska M, Rucinski M, Ziolkowska A, et al. Enucleation-induced rat adrenal gland regeneration: expression profile of selected genes involved in control of adrenocortical cell proliferation. Int J Endocrinol. 2014; 2014: 130359.
- Albertin G, Rucinski M, Carraro G, et al. Adrenomedullin and vascular endothelium growth factor genes are overexpressed in the regenerating rat adrenal cortex, and AM and VEGF reciprocally enhance their mRNA expression in cultured rat adrenocortical cells. Int J Mol Med. 2005; 16(3): 431–435.
- Trejter M, Neri G, Rucinski M, et al. Neuromedin-U stimulates enucleation-induced adrenocortical regeneration in the rat. Int J Mol Med. 2008; 21(6): 683–687.
- Ziolkowska A, Spinazzi R, Albertin G, et al. Orexins stimulate glucocorticoid secretion from cultured rat and human adrenocortical cells, exclusively acting via the OX1 receptor. J Steroid Biochem Mol Biol. 2005; 96(5): 423–429.
- Rucinski M, Ziolkowska A, Szyszka M, et al. Cerebellin and des-cerebellin exert ACTH-like effects on corticosterone secretion and the intracellular signaling pathway gene expression in cultured rat adrenocortical cells--DNA microarray and QPCR studies. Int J Mol Med. 2009; 23(4): 539–546.
- Rucinski M, Ziolkowska A, Tyczewska M, et al. Expression of prepro-ghrelin and related receptor genes in the rat adrenal gland and evidences that ghrelin exerts a potent stimulating effect on corticosterone secretion by cultured rat adrenocortical cells. Peptides. 2009; 30(8): 1448–1455.
- Yu Y, Fuscoe JC, Zhao C, et al. A rat RNA-Seq transcriptomic BodyMap across 11 organs and 4 developmental stages. Nat Commun. 2014; 5: 3230.
- Tortorella C, Macchi C, Spinazzi R, et al. Ghrelin, an endogenous ligand for the growth hormone-secretagogue receptor, is expressed in the human adrenal cortex. Int J Mol Med. 2003; 12(2): 213–217.
- Rucinski M, Albertin G, Spinazzi R, et al. Cerebellin in the rat adrenal gland: gene expression and effects of CER and [des-Ser1]CER on the secretion and growth of cultured adrenocortical cells. Int J Mol Med. 2005; 15(3): 411–415.
- Rucinski M, Tortorella C, Ziolkowska A, et al. Steroidogenic acute regulatory protein gene expression, steroid-hormone secretion and proliferative activity of adrenocortical cells in the presence of proteasome inhibitors: In vivo studies on the regenerating rat adrenal cortex. Int J Mol Med. 2008; 21(5): 593–597.
- Gentleman RC, Carey VJ, Bates DM, et al. Bioconductor: open software development for computational biology and bioinformatics. Genome Biol. 2004; 5(10): R80.
- Benjamini Y, Hochberg Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. Journal of the Royal Statistical Society Series B (Methodological). 1994; 57(1): 289–300.
- Uhlén M, Björling E, Agaton C, et al. A human protein atlas for normal and cancer tissues based on antibody proteomics. Mol Cell Proteomics. 2005; 4(12): 1920–1932.
- Cohen HY, Miller C, Bitterman KJ, et al. Calorie restriction promotes mammalian cell survival by inducing the SIRT1 deacetylase. Science. 2004; 305(5682): 390–392.
- Yu W, Zhou HF, Lin RB, et al. Short‑term calorie restriction activates SIRT1‑4 and ‑7 in cardiomyocytes in vivo and in vitro. Mol Med Rep. 2014; 9(4): 1218–1224.
- Miyazaki S, Kakutani K, Yurube T, et al. Recombinant human SIRT1 protects against nutrient deprivation-induced mitochondrial apoptosis through autophagy induction in human intervertebral disc nucleus pulposus cells. Arthritis Res Ther. 2015; 17: 253.
- Novoa-Herran S, Umaña-Perez A, Canals F, et al. Serum depletion induces changes in protein expression in the trophoblast-derived cell line HTR-8/SVneo. Cell Mol Biol Lett. 2016; 21: 22.
- Rahmani M, Khorasani HR, Golpour M, et al. Stable down-regulation of HLA class-I by serum starvation in human PBMCs. Iran J Immunol. 2016; 13(1): 54–63.
- Tong J, Sun D, Yang C, et al. Serum starvation and thymidine double blocking achieved efficient cell cycle synchronization and altered the expression of p27, p53, bcl-2 in canine breast cancer cells. Res Vet Sci. 2016; 105: 10–14.
- Hasmann M, Schemainda I. FK866, a highly specific noncompetitive inhibitor of nicotinamide phosphoribosyltransferase, represents a novel mechanism for induction of tumor cell apoptosis. Cancer Res. 2003; 63(21): 7436–7442.
- Li D, Dammer EB, Sewer MB. Resveratrol stimulates cortisol biosynthesis by activating SIRT-dependent deacetylation of P450scc. Endocrinology. 2012; 153(7): 3258–3268.
- Marti N, Bouchoucha N, Sauter KS, et al. Resveratrol inhibits androgen production of human adrenocortical H295R cells by lowering CYP17 and CYP21 expression and activities. PLoS One. 2017; 12(3): e0174224.
- Fu H, Wada-Hiraike O, Hirano M, et al. SIRT3 positively regulates the expression of folliculogenesis- and luteinization-related genes and progesterone secretion by manipulating oxidative stress in human luteinized granulosa cells. Endocrinology. 2014; 155(8): 3079–3087.
- Li S, Banck M, Mujtaba S, et al. p53-induced growth arrest is regulated by the mitochondrial SirT3 deacetylase. PLoS One. 2010; 5(5): e10486.
- Xiao K, Jiang J, Wang W, et al. Sirt3 is a tumor suppressor in lung adenocarcinoma cells. Oncol Rep. 2013; 30(3): 1323–1328.
- Jeong SM, Lee A, Lee J, et al. SIRT4 protein suppresses tumor formation in genetic models of Myc-induced B cell lymphoma. J Biol Chem. 2014; 289(7): 4135–4144.
