Animal Reproduction (AR)
Animal Reproduction (AR)
Original Article

A Brazilian pulp and paper mill effluent disrupts energy metabolism in immature rat testis and alters Sertoli cell secretion and mitochondrial activity

Vanessa Staldoni de Oliveira; Allisson Jhonatan Gomes Castro; Juliana Tonietto Domingues; Ariane Zamoner Pacheco de Souza; Débora da Luz Scheffer; Alexandra Latini; Carlos Henrique Lemos Soares; Glen Van Der Kraak; Fátima Regina Mena Barreto Silva

Downloads: 0
Views: 49


Abstract: Our objective was to investigate whether the pulp and paper mill industry effluent could affect the testis and Sertoli cells in a fast exposure period. For this, the present study was carried out in immature rats at 10-day-old. Testis treated in vitro with 4% effluent for 1 h presented changes in energy metabolism in terms of a decrease in lactate content and glucose uptake. Elevation in GSH content, as an antioxidant defense mechanism, was also detected. Sertoli cells treated with 4% effluent for 1 hour showed alterations in the mitochondrial metabolism that favor the decoupling of oxidative phosphorylation and the generation of oxygen reactive species and also a time and concentration-dependent delay secretion of acidic vesicles. Our results showed that pollutants present in the pulp and paper mill effluents, in a short time of exposure, are capable of inducing alterations in important metabolic functions in the testis and in Sertoli cells that are crucial for the correct progression of spermatogenesis and fertility.


testis, Sertoli, effluent, lactate, secretion


Agarwal A, Virk G, Ong C, Du Plessis SS. Effect of oxidative stress on male reproduction. World J Mens Health. 2014;32(1):1-17. PMid:24872947.

Aitken RJ, Gibb Z, Baker MA, Drevet J, Gharagozloo P. Causes and consequences of oxidative stress in spermatozoa. Reprod Fertil Dev. 2016;28(1-2):1-10. PMid:27062870.

Aitken RJ, Roman SD. Antioxidant systems and oxidative stress in the testes. In: Cheng CY, editor. Molecular mechanisms in spermatogenesis. Austin: Landes Bioscience/Springer Science; 2008. p. 154-71.

Alves MG, Martins AD, Rato L, Moreira PI, Socorro S, Oliveira PF. Molecular mechanisms beyond glucose transport in diabetes-related male infertility. Biochim Biophys Acta. 2013;1832(5):626-35. PMid:23348098.

Bauché F, Fouchard MH, Jégou B. Antioxidant system in rat testicular cells. FEBS Lett. 1994;349(3):392-6. PMid:8050602.

Bergmeyer HU. Methods of enzymatic analysis. 3rd ed. Deerfield Beach: Verlag Chemie; 1983. Vol. 3, p. 118-25.

Beutler E, Duron O, Kelly BM. Improved method for the determination of blood glutathione. J Lab Clin Med. 1963;61:882-90. PMid:13967893.

Bird RP, Draper HH. Comparative studies on different methods of malonaldehyde determination. Methods Enzymol. 1984;105:299-305. PMid:6727668.

Bisht S, Faiq M, Tolahunase M, Dada R. Oxidative stress and male infertility. Nat Rev Urol. 2017;14(8):470-85. PMid:28508879.

Blomberg Jensen M. Vitamin D and male reproduction. Nat Rev Endocrinol. 2014;10(3):175-86. PMid:24419359.

Burtis CA, Ashwood ER, Bruns DE. Tietz fundamentals of clinical chemistry. 6th ed. St. Louis: Saunders Elsevier; 2007.

Castro AJG, Baptista IE, Moura KRS, Padilha F, Tonietto J, de Souza AZP, Soares CHL, Silva FRMB, Van Der Kraak G. Exposure to a Brazilian pulp mill effluent impacts the testis and liver in the zebrafish. Comp Biochem Physiol C Toxicol Pharmacol. 2018;206-207:41-7. PMid:29499384.

Cavalli VLLO, Cattani D, Rieg CEH, Pierozan P, Zanatta L, Parisotto EB, Wilhelm Filho D, Silva FRMB, Pessoa-Pureur R, Zamoner A. Roundup disrupts male reproductive functions by triggering calcium-mediated cell death in rat testis and Sertoli cells. Free Radic Biol Med. 2013;65:335-46. PMid:23820267.

Celino FT, Yamaguchi S, Miura C, Ohta T, Tozawa Y, Iwai T, Miura T. Tolerance of spermatogonia to oxidative stress is due to high levels of Zn and Cu/Zn superoxide dismutase. PLoS One. 2011;6(2):e16938. PMid:21364994.

Chiang G, Barra R, Diaz-Jaramillo M, Rivas M, Bahamonde P, Munkittrick KR. Estrogenicity and intersex in juvenile rainbow trout (Oncorhynchus mykiss) exposed to Pine/Eucalyptus pulp and paper production effluent in Chile. Aquat Toxicol. 2015;164:126-34. PMid:25956323.

Corrêa de Deus JJ, Bagatini KP. Conscientização ambiental: conservação da flora e da fauna do Parque Estadual Rio Canoas, Campos Novos, Santa Catarina. Unoesc & Ciência. 2016;7:153-8.

Costa RR, Varanda WA, Franci CR. A calcium-induced calcium release mechanism supports luteinizing hormone-induced testosterone secretion in mouse Leydig cells. Am J Physiol Cell Physiol. 2010;299(2):C316-23. PMid:20519450.

De Paula Martins R, Glaser V, Scheffer DL, De Paula Ferreira PM, Wannmacher CMD, Farina M, Oliveira PA, Prediger RD, Latini A. Platelet oxygen consumption as a peripheral blood marker of brain energetics in a mouse model of severe neurotoxicity. J Bioenerg Biomembr. 2013;45(5):449-57. PMid:23471523.

Dey S, Choudhury MD, Das S. A review on toxicology of paper mill effluent on fish. Bull Environ Pharmacol. Life Sci. 2013;2:17-23.

Dorrington JH, Roller NF, Fritz IB. Effects of follicle-stimulating hormone on cultures of Sertoli cell preparations. Mol Cell Endocrinol. 1975;3(1):57-70. PMid:168104.

Erkkilä K, Aito H, Aalto K, Pentikainen V, Dunkel L. Lactate inhibits germ cell apoptosis in the human testis. Mol Hum Reprod. 2002;8(2):109-17. PMid:11818513.

França LR, Hess RA, Dufour JM, Hofmann MC, Griswold MD. The Sertoli cell: one hundred fifty years of beauty and plasticity. Andrology. 2016;4(2):189-212. PMid:26846984.

Galardo MN, Riera MF, Pellizzari EH, Chemes HE, Venara MC, Cigorraga SB, Meroni SB. Regulation of expression of Sertoli cell glucose transporters 1 and 3 by FSH, IL1, and bFGF at two different time-points in pubertal development. Cell Tissue Res. 2008;334(2):295-304. PMid:18802725.

Gonçalves R, Zamoner A, Zanatta L, Zanatta AP, Remor AP, Scheffer DL, Latini A, Silva FRMB. 1,25(OH)2 vitamin D3 signalling on immature rat Sertoli cells: gamma-glutamyl transpeptidase and glucose metabolism. J Cell Commun Signal. 2017;11(3):233-43. PMid:28160135.

Gonçalves R, Zanatta AP, Cavalari FC, Nascimento MAW, Delalande-Lecapitaine C, Bouraïma-Lelong H, Silva FRMB. Acute effect of bisphenol A: signaling pathways on calcium influx inimmature rat testes. Reprod Toxicol. 2018;77:94-102. PMid:29476780.

Habig WH, Pabst MJ, Jakoby WB. Glutathione-S-transferases: the first enzymatic step in mercapturic acid formation. J Biol Chem. 1974;249(22):7130-9. PMid:4436300.

Hall A, Larsen AK, Parhamifar L, Meyle KD, Wu LP, Moghimi SM. High resolution respirometry analysis of polyethylenimine mediate mitochondrial energy crisis and cellular stress: mitochondial proton leak and inhibition of the electron transport system. Biochim Biophys Acta. 2013;1827(10):1213-25. PMid:23850549.

Ham J, Lim W, Park S, Bae H, You S, Song G. Synthetic phenolic antioxidant propyl gallate induces male infertility through disruption of calcium homeostasis and mitochondrial function. Environ Pollut. 2019;248:845-56. PMid:30856500.

Hutter E, Renner K, Pfister G, Stöckl P, Jansen-Dürr P, Gnaiger E. Senescence-associated changes in respiration and oxidative phosphorylation in primary human fibroblasts. Biochem J. 2004;380(3):919-28. PMid:15018610.

Ischiropoulos H, Gow A, Thom SR, Kooy NY, Royall JA, Crow JP. detection of reactive nitrogen species using 2,7- dichlorodihydrofluorescein and dihydrorhodamine 123. Methods Enzymol. 1999;301:367-73. PMid:9919585.

Jutte NH, Grootegoed J, Rommerts F, Van Der Molen H. Exogenous lactate is essential for metabolic activities in isolated rat spermatocytes and spermatids. J Reprod Fertil. 1981;62(2):399-405. PMid:7252921.

Kappel VD, Zanatta L, Postal BG, Silva FRMB. Rutin potentiates calcium uptake via voltage-dependent calcium channel associated with stimulation of glucose uptake in skeletal muscle. Arch Biochem Biophys. 2013;532(2):55-60. PMid:23395857.

Liu B, Zhang X, Qin M, Song A, Cui T, Wang C. Malfunction of Sertoli cell secretion leads to testicular damage in diabetic rats. Int J Clin Exp Med. 2016;9:9316-21.

Liu L, Chang X, Zhang Y, Wu C, Li R, Tang L, Zhou Z. Fluorochloridone induces primary cultured Sertoli cells apoptosis: involvement of ROS and intracellular calcium ions-mediated ERK1/2 activation. Toxicol In Vitro. 2018;47:228-37. PMid:29248592.

Lopes MS, Veettil BK, Saldanha DL. Assessment of small-scale ecosystem conservation in the Brazilian Atlantic Forest: a study from Rio Canoas State Park, Southern Brazil. Sustainability. 2019;11(10):2948.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent. J Biol Chem. 1951;193(1):265-75. PMid:14907713.

Lyon K, Adams A, Piva M, Asghari P, Moore ED, Vogl AW. Ca2+ signaling machinery is present at intercellular junctions and structures associated with junction turnover in rat Sertoli cells. Biol Reprod. 2017;96(6):1288-302. PMid:28486663.

Mendive F, Laurent P, Van Schoore G, Skarnes W, Pochet R, Vassart G. Defective postnatal development of the male reproductive tract in LGR4 knockout mice. Dev Biol. 2006;290(2):421-34. PMid:16406039.

Misra HP, Fridovich I. The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem. 1972;247(10):3170-5. PMid:4623845.

Morales A, Mohamed F, Cavicchia JC. Apoptosis and blood-testis barrier during the first spermatogenic wave in the pubertal rat. Anat Rec. 2007;290(2):206-14. PMid:17441213.

Mruk DD, Cheng CY. The mammalian blood-testis barrier: its biology and regulation. Endocr Rev. 2015;36(5):564-91. PMid:26357922.

Orlowski M, Meister A. γ-Glutamyl-p-nitroanilide: a new convenient substrate for determination and study of l- and d-γ-glutamyltranspeptidase activities. Biochim Biophys Acta. 1963;73:679-81. PMid:14074151.

Paraidathathu T, De Groot H, Kehrer JP. Production of reactive oxygen by mitochondria from normoxic and hypoxic rat heart tissue. Free Radic Biol Med. 1992;13(4):289-97. PMid:1383097.

Picut CA, Parker GA. Postnatal organ development as a complicating factor in juvenile toxicity studies in rats. Toxicol Pathol. 2017;45(1):248-52. PMid:27753635.

Rato L, Alves MG, Socorro S, Duarte AI, Cavaco JE, Oliveira PF. Metabolic regulation is important for spermatogenesis. Nat Rev Urol. 2012;9(6):330-8. PMid:22549313.

Reis MMS, Moreira AC, Sousa M, Mathur PP, Oliveira PF, Alves MG. Sertoli cell as a model in male reproductive toxicology: advantages and disadvantages. J Appl Toxicol. 2015;35(8):870-83. PMid:25693974.

Riera MF, Galardo MN, Pellizzari EH, Meroni SB, Cigorraga SB. Molecular mechanisms involved in Sertoli cell adaptation to glucose deprivation. Am J Physiol Endocrinol Metab. 2009;297(4):E907-14. PMid:19638510.

Rosso A, Pansera M, Zamoner A, Zanatta L, Bouraïma-Lelong H, Carreau S, Silva FRMB. 1α,25(OH)2-Vitamin D3 stimulates rapid plasma membrane calcium influx via MAPK activation in immature rat Sertoli cells. Biochimie. 2012;94(1):146-54. PMid:22015633.

Sedha S, Kumar S, Shukla S. Role of oxidative stress in male reproductive dysfunctions with reference to phthalate compounds. Urol J. 2015;12(5):2304-16. PMid:26571312.

Smith LB, Milne L, Nelson N, Eddie S, Brown P, Atanassova N, O’Bryan MK, O’Donnell L, Rhodes D, Wells S, Napper D, Nolan P, Lalanne Z, Cheeseman M, Peters J. KATNAL1 regulation of sertoli cell microtubule dynamics is essential for spermiogenesis and male fertility. PLoS Genet. 2012;8(5):e1002697. PMid:22654668.

Waye A, Lado WE, Martel PH, Arnason JT, Trudeau VL. Ovulation but not miltproduction is inhibited in fathead minnows (Pimephales promelas) exposed to a reproductively inhibitory pulp mill effluent. Reprod Biol Endocrinol. 2014;12(1):43. PMid:24884628.

Zanatta AP, Zanatta L, Gonçalves R, Zamoner A, Silva FRMB. Rapid responses to reverse T3 hormone in immature rat Sertoli cells: calcium uptake and exocytosis mediated by integrin. PLoS One. 2013;8(10):e77176. PMid:24130850.

Submitted date:

Accepted date:

5eff246f0e8825da09f8e224 animreprod Articles
Links & Downloads

Anim Reprod

Share this page
Page Sections