[en] Metformin, an insulin sensitiser from the biguanide family of molecules, is used for the treatment of insulin resistance in type 2 diabetes individuals. It increases peripheral glucose uptake and may reduce food intake. Based on the tight link between metabolism and fertility, we investigated the role of metformin on testicular function using in vitro culture of Sertoli cells and seminiferous tubules, complemented by in vivo data obtained following metformin administration to prepubertal chickens. In vitro, metformin treatment reduced Sertoli cell proliferation without inducing apoptosis and morphological changes. The metabolism of Sertoli cells was affected because lactate secretion by Sertoli cells increased approximately twofold and intracellular free ATP was negatively impacted. Two important pathways regulating proliferation and metabolism in Sertoli cells were assayed. Metformin exposure was not associated with an increased phosphorylation of AKT or ERK. There was a 90% reduction in the proportion of proliferating germ cells after a 96-h exposure of seminiferous tubule cultures to metformin. In vivo, 6-week-old chickens treated with metformin for 3 weeks exhibited reduced testicular weight and a 50% decrease in testosterone levels. The expression of a marker of undifferentiated germ cells was unchanged in contrast to the decrease in expression of 'protamine', a marker of differentiated germ cells. In conclusion, these results suggest that metformin affects the testicular energy content and the proliferative ability of Sertoli and germ cells.
Disciplines :
Agriculture & agronomy
Author, co-author :
Faure, Mélanie ; Université de Liège - ULiège > GIGA > GIGA Neurosciences - Neuroendocrinology ; INRAUMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly,
Guibert, E; INRAUMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly,
Alves, S; INRAUMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly,
Pain, B; INSERMUMR846, Sterm Cell and Brain Research Institute, Bron, France INRAUSC1361,
Ramé, C; INRAUMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly,
Dupont, J; INRAUMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly,
Brillard, J P; FERTIL'AVI37360, Rouziers de Touraine, France.
Froment, P; INRAUMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly,
Language :
English
Title :
The insulin sensitiser metformin regulates chicken Sertoli and germ cell populations.
Alimova IN, Liu B, Fan Z, Edgerton SM, Dillon T, Lind SE & Thor AD 2014 Metformin inhibits breast cancer cell growth, colony formation and induces cell cycle arrest in vitro. Cell Cycle 8 909-9 15. (doi:10.4161/cc.8.6.7933)
Alves MG, Martins AD, Vaz CV, Correia S, Moreira PI, Oliveira PF & Socorro S 2014 Metformin and male reproduction: effects on Sertoli cell metabolism. British Jour nal of Pharmacology 171 1033-1042. (doi:10.1111/bph.12522)
Bertoldo MJ, Guibert E, Tartarin P, Guillory V & Froment P 2014a Effect of metformin on the fertilizing ability of mouse spermatozoa. Cryobiology 68 262-268. (doi:10.1016/j.cryobiol.2014.02.006)
Bertoldo MJ, Faure M, Dupont J & Froment P 2014b Impact of metformin on reproductive tissues : an overview from gametogenesis to gestation. Annals of Translational Medicine 2 1-13. (doi:10.1016/j. mce.2016.01.001)
Bhasin S, Woodhouse L & Storer TW 2001 Proof of the effect of testosterone on skeletal muscle. Journal of Endocrinology 170 27-38. (doi:10.1111/jgs.2008.56.issue-11)
Bilodeau-Goeseels S, Magyara N & Collignon C 2014 Characterization of the effects of metformin on porcine oocyte meiosis and on AMPactiv ated protein kinase activation in oocytes and cumulus cells. Zygote 22 275-85. (doi:10.1017/S0967199413000075)
Blomqvist A, Berg C, Holm L, Brandt I, Ridderstrale Y & Brunstrom B 2006 Defective reproductive organ morphology and function in domestic rooster embryonically exposed to o-p'-DDT or ethynylestradiol. Biology of Reproduction 74 481-486. (doi:10.1897/05-619R.1)
Borland K, Mita M, Oppenheimer CL, Blinderman LA, Massague J, Hall PF & Czech MP 1984 The actions of insulin-like growth factors I and II on cultured Sertoli cells. Endocrinology 114 240-246. (doi:10.1210/endo-114-1-240)
Brilla rd JP & De Reviers M 1981 Testis development and daily sperm output in guinea-fowl raised under constant daily photoperiods. Reproduction, Nutrition, Developpement 21 1105-1112. (doi:10.1051/rnd:19810809)
Buz zai M, Jones RG, Amaravadi RK, Lum JJ, DeBerardinis RJ, Zhao F, Viollet B & Thompson CB 2007 Systemic treatment with the antidiabetic drug metformin selectively impairs p53-deficient tumor cell growth. Cancer Research 67 6745-6752. (doi:10.1158/0008-5472.CAN-06-4447)
Chen W-L, Wei H-W, Chiu W-Z, Kang C-H, Lin T-H, Hung C-C, Chen MC, Shieh M-S, Lee C-C & Lee H-M 2011 Metformin regul ates hepatic lipid metabolism through activating AMP-Activated protein kinase and inducing ATGL in laying hens. European Journal of Pharmacology 671 107-112. (doi:10.1016/j.ejphar.2011.09.029)
Cheung PCF, Salt IP, Davies SP, Hardie DG & Carling D 2000 their role in AMP binding. Biochemistry 346 659-669. (doi:10.1042/bj3460659)
Chou SH-H & Mantzoros C 2014 Leptin in human reproductive disorders. Journal of Endocrinology 223 T49-T62.
Coyral-Castel S, Tosca L, Ferreira G, Jeanpierre E, Rame C, Lomet D, Caraty A, Monget P, Chabrolle C & Dupont J 2008 The effect of AMPactivated kinase activation on gonadotrophin-releasing hormone s ecretion in GT1-7 cells and its potential role in hypothalamic regulation of the oestrous cyclicity in rats. Journal of Neuroendocrinology 20 335-346. (doi:10.1111/jne.2008.20.issue-3)
Crepieux P, Marion S, Mar tinat N, Fafeur V, Vern YL, Kerboeuf D, Guillou F & Reiter E 2001 The ERK-dependent signalling is stagespecifically modulated by FSH, during primary Sertoli cell maturation. Oncogene 20 4696-4709. (do i:10.1038/sj.onc.1204632)
De Reviers M 1971 Le developpement testiculaire chez le coq. Annales de biologie animale, biochimie, biophysique 11 519-530. (doi:10.1051/rnd:19710401)
De Reviers M, Hochereau-de Reviers MT, Blanc MR, Brillard JP, Courot M & Pelletier J 1980 Control of Sertoli and germ cell populations in the cock and sheep testes. Reproduction, Nutrition, Développement 20 241-9. (doi:10.1051/rnd:19800206)
Djakiew D & Dym M 1988 Pachytene spermatocyte protein influence Sertoli cell function. Biological Research for Nursing 39 1193-1205. (doi:10.1158/1541-7786.MCR-10-0342)
Dupont J,Metayer-Coustard S,Ji B,Rame C,Gespach C,Voy B,Simon J 2012 Characterization of major elements of insulin signaling cascade in chicken adipose tissue: apparent insulin refractoriness General and Co mparative Endocrinology 176 86-93 10.1016/j. ygcen.2011.12.030)
Fernandez-Fernandez R, Martini AC, Navarro VM, Castellano JM, Dieguez C, Aguilar E, Pinilla L & Tena-Sempere M 2006 Novel signals for the integration of energy balance and reproduction. Molecular and Cellular Endocrinology 254-255 127-132. (doi:10.1016/j. mce.2006.04.026)
Franks S, Robinson S & W illis DS 1996 Nutrition, insulin and polycystic ovary syndrome. Reviews of Reproduction 1 47-53. (doi:10.1046/j.1365-2265.2000.01123.x)
Froment P, Staub C, Hembert S, Pisselet C, Magistrini M, Delaleu B, Seurin D, Levine JE, Johnson L, Binoux M et al. 2004 Reproductive abnormalities in human insulin-like growth factor-binding protein-1 transgenic male mice. Endocrinology 145 2080-2091. (doi :10.1210/en.2003-0956)
De Gendt K 2004 A Sertoli cell-selective knockout of the androgen receptor causes spermatogenic arrest in meiosis. PNAS 101 1327-1332. (doi:10.1073/pnas.0308114100)
Galar do MN, Riera MF, Pellizzari EH, Cigorraga SB & Meroni SB 2007 The AMP-Activated protein kinase activator, 5-Aminoimidazole-4-carboxamide-1-b-d-ribonucleoside, regulates lactate production in rat Sertoli cells. Journal of Molecular Endocrinology 39 279-288. (doi:10.1016/j.ygcen.2015.08.011)
Guibert E, Briere S, Pelletier R, Brillard JP & Froment P 2011 Characterization of chicken Sertoli cells in vitro. Poult ry Science 90 1276-1286. (doi:10.3382/ps.2010-01081)
Guibert E, Prieur B, Cariou R, Courant F, Antignac JP, Pain B, Brillard JP & Froment P 2013 Effects of mono-(2-ethylhexyl) phthalate (MEHP) on chicken germ cells cultured in vitro. Environmental Science and Pollution Research International 20 2771-2783. (doi:10.1007/s11356-013-1487-2)
Havenstein GB, Ferket PR & Qureshi MA 2003 Growth, livability, and feed conversion of 1957 versus 2001 broilers when fed representative 1957 and 2001 broiler diets. Poultry Science 82 1500-1508. (doi:10.1 093/ps/86.2.241)
Hochereau-de Reviers MT, Perreau C, Pisselet C, Fontaine I & Monet-Kuntz C 1990 Comparisons of endocrinological and testis parameters in 18-month-old Ile de France and Romanov rams. Domesti c Animal Endocrinology 7 63-73. (doi:10.1677/joe.0.1310101)
Hocking PM 1987 Nutritional interactions with reproduction in birds. Proceedings of the Nutrition Society 46 217-225. (doi:10.1079/PNS19870029)
Holsberger DR, Buchold GM, Leal MC, Kiesewetter SE, O'Brien DA, Hess RA, Franca LR, Kiyokawa H & Cooke PS 2005 Cell-cycle inhibitors p27Kip1 and p21Cip1 regulate murine Sertoli cell p roliferation. Biology of Reproduction 72 1429-1436. (doi:10.1095/biolreprod.105.040386)
Ibáñ ez L, Ong K, Valls C, Marcos MV, Dunger DB & de Zegher F (2006) Metformin treatment to preven t early puberty in girls with precocious pubarche. Journal of Clinical Endocrinology and Metabolism 91 2888-2891. (doi:10.1210/jc.2006-0336)
Ji B, Ernest B, Gooding JR, Das S, Saxton AM, Simon J, Dupont J, Metayer-Coustard S, Campagna SR & Voy BH 2012 Transcriptomic and metabolomic profiling of chicken adipose tissue in response to insulin neutralization and fasting. BMC Genomics 13 441. (doi:10.118 6/1471-2164-13-441)
Ji B, Middleton JL, Ernest B, Saxton AM, Lamont SJ, Campagna SR & Voy BH 2014 Molecular and metabolic profiles suggest that increased lipid catabolism in adipose tissue contributes to leanness in domestic chickens. Physiological Genomics 46 315-327. (doi:10.1152/physiolgenomics.00163.2013)
Jutte NH, Grootegoed JA, Rommerts FFG & van der Molen HJ 1981 Exogenous lactate is ess ential for metabolic activities in isolated rat spermatocytes and spermatids. Journal of Reproduction and Fertility 62 399-405. (doi:10.1530/jrf.0.0620399)
Khan SA, Ndjountche L, Pratchard L, Spicer LJ & Davis JS 2002 Folliclestimulating hormone amplifies insulin-like growth factor I-mediated activation of AKT/protein kinase B signaling in immature rat Sertoli cells. Endocrinology 143 2259-2267. (doi:10.1210/endo.143.6 .8838)
Lavial F, Acloque H, Bachelard E, Nieto MA, Samarut J & Pain B 2009 Ectopic expression of Cvh (chicken Vasa homologue) mediates the reprogramming of chicken embryonic stem cells to a germ cell fate. Developmental Biology 330 73-82. (doi:10.1016/j.ydbio.2009.03.012)
Legendre A, Froment P, Desmots S, Lecomte A, Habert R & Lem azurier E 2010 An engineered 3D blood-testis barrier model for the assessment of reproductive toxicity potential. Biomaterials 31 4492-4505. (doi:10.1016/j.biomaterials.2010.02.029)
Leverve XM,Guigas B,Detaille D,Batandier C,Koceir EA,Chauvin C,Fontaine E,Wiernsperger NF 2003 Mitochondrial metabolism and type-2 diabetes: a specific target of metformin Diabetes & Metabolism 29 6S88-6S94.
Migliorini D, Denchi EL, Danovi D, Jochemsen A, Capillo M, Gobbi A, Helin K, Pelicci PG & Marine J-C 2002 Mdm4 (Mdmx) regulates p53-induced growth arrest and neuronal cell death during early embryonic mouse dev elopment. Molecular and Cellular Biology 22 5527-5538. (doi:10.1128/MCB.22.15.5527-5538.2002)
Nguyen TMD, Alves S, Grasseau I, Metayer-Coustard S, Praud C, Froment P & Blesbois E 2014 Central role of 5'-AMP-Activated protein kinase in chicken sperm functions. Biology of Reproduction 91 121. (doi:10.1095/biolreprod.114.121855)
Oonk RB, Jansen R & Grootegoed JA 1989 Differential effects of follicle stimulating hormone, insulin, and insulin-like growth factor I on hexose uptake and lactate production by rat Sertoli cell s. Journal of Cellular Physiology 139 210-218. (doi:10.1002/(ISSN)1097-4652)
Pasquali R 2006 Obesity, fat distribution and infertility. Maturitas 54 363-371. (doi:10.1016/j.maturitas.2006.04.018)
Petersen C & Soder O 2006 The Sertoli cell-A hormonal target and 'super'nurse for germ cells that determines testicular size. Hormone Research 66 153-161. (doi:10.1159/000094142)
Pirke KM, Bofilias I, Spyra B, Langhammer H & Pabst HW 1982 Capillary blood flow in the tests and testosterone secretion in the starved rat. Experientia 38 516-517. (doi:10.1007/BF01952671)
Rato L, Alves MG, Cavaco JE & Oliveira PF 2014 High-energy diets: a threat for male fertility? Obesity Reviews 15 996-1007. (doi:10.1016/j. biocel.2015.07.001)
Rena G, Pearson ER & Sakamoto K 2013 Molecular mechanism of action of metformin: old or new insights? Diabetologia 56 1898-1906. (doi:10.1007/s00125-013-2991-0)
Rengaraj D, Zheng YH, Kang KS, Park KJ, Lee BR, Lee SI, Choi JW & Han JY 2010 Conserved expression pattern of chicken DAZL in primordial germ cells and germ-line cells. Theriogenology 74 765-776. (doi:10.1016/j. theriogenology.2010.04.001)
Rice S, Pellatt L, Ramanathan K, Whitehead SA & Mason HD 2009 Metformin inhibits aromatase via an extracellular signal-regulated kinase-mediated pathway. Endocrinology 150 4794-4801. (doi:10 .1210/en.2009-0540)
Schneider JE 2004 Energy balance and reproduction. Physiology & Behavior 81 289-317. (doi:10.1016/j.physbeh.2004.02.007)
Sedqyar M, Weng Q, Watanabe G, Kandi el MM, Takahashi S, Suzuki AK, Taneda S & Taya K 2008 Secretion of inhibin in male Japanese quail (Coturnix japonica) from one week of age to sexual maturity. Journal of Reproduction and Development 54 100-106.(doi:10.1262/jrd.19155)
Staub C, Hue D, Nicolle JC, Perrard-Sapori MH, Segretain D & Durand P 2000 The whole meiotic process can occur in vitro in untransformed rat spermatogenic cells. Experiment al Cell Research 260 85-95. (doi:10.1006/excr.2000.4998)
Svechnikov K, Spatafora C, Svechnikova I, Tringali C & Soder O 2009 Effects of resveratrol analogs on steroidogenesis and mitochondrial f unction in rat Leydig cells in vitro. Journal of Applied Toxicology 29 673-680. (doi:10.1002/jat.v29:8)
Tan K AL, Turner KJ, Saunders PTK, Verhoeven G, De Gendt K, Atanassova N & Sharpe RM 2005 Androgen regulation of stage-dependent cyclin D2 expression in Sertoli cells suggests a role in modulating androgen action on spermatog enesis. Biology of Reproduction 72 1151-1160. (doi:10.1095/biolreprod.104.037689)
Tartarin P, Guibert E, Toure A, Ouiste C, Leclerc J, Sanz N, Briere S, Dacheux J-L, Delaleu B, McNeilly JR et al. 2012 a Inactivation of AMPK?1 induces asthenozoospermia and alters spermatozoa morphology. Endocrinology 153 3468-3481. (doi:10.1210/en.2011-1911)
Tartarin P, Moison D, Guibert E, Dupont J, Habert R, Rouiller-Fabre V, Frydman N, Pozzi S, Frydman R, Lecureuil C et al. 2012 b Metformin exposure affects human and mouse fetal testicular cells. Human Reproduction 27 3304-3314. (doi:10.1093/humrep/des264)
Tosca L, Crochet S, Ferre P, Foufelle F, Tesseraud S & Dupont J 2006 AMPactivated protein kinase activation modulates progesterone secretion in granulosa cells from hen preovulatory follicles. Journal of Endocrinology 190 85-97. (doi:10.1677/joe.1.06828)
Tosca L, Uzbekova S, Chabrolle C & Dupont J 2007 Possible role of 5'AMP-Activated protein kinase in the metformin-mediat ed arrest of bovine oocytes at the germinal vesicle stage during in vitro maturation. Biology of Reproduction 77 452-465. (doi:10.1095/biolreprod.107.060848)
Tosca L,Froment P,Rame C,McNeilly JR,McNeilly AS,Maillard V,Dupont J 2011 Metformin decreases GnRH-And activininduced gonadotropin secretion in rat pituitary cells: potential involvement of adenosine 5'monophosphate-Activated protein kinase (PRKA Biology of Reproduction 84 351-362 10.1095/biolreprod.110.087023)
Tsunekawa N, Naito M, Sakai Y, Nishida T & Noce T 2000 Isolation of chicken Vasa homolog gene and tracing the origin of primordial germ cel ls. Development 127 2741-2750.
Walzem RL & Chen S 2014 Obesity-induced dysfunctions in female reproduction: lessons from birds and mammals 1-3. American Society for Advances in Nutrition 5 199-206. (doi:10.1016/j.anireprosci. 2016.02.002)
Yan W, Mu Y, Yu N, Yi T, Zhang Y, Pang X, Cheng D & Yang J (2015) Protective effects of metformin on reproductive function in obese m ale rats induced by high-fat diet. Journal of Assisted Reproduction and Genetics 32 1097-1104. (doi:10.1007/s10815-015-0506-2)
Zang M, Xu S, Maitland-Toolan KA, Zuccollo A, Hou X, Jiang B, Wierz bicki M, Verbeuren TJ & Cohen RA 2006 Polyphenols stimulate AMP-Activated protein kinase, lower lipids, and inhibit accelerated atherosclerosis in diabetic LDL receptor-deficient mice. Diabetes 55 2180-2191. (doi :10.2337/db05-1188)