Abstract
Perfluorooctanoic acid (PFOA) is correlated with male reproductive dysfunction in animals and humans, but the underlying mechanisms for this remain unknown. To explore the potential reproductive toxicity of PFOA, we studied blood–testis barrier (BTB) damage using in vivo and in vitro models. Male mice were gavage-administered PFOA (0–20 mg/kg/d) for 28 consecutive days, and breeding capacity and permeability of the Sertoli cell-based BTB were estimated. Primary Sertoli cells (SCs) were exposed to PFOA (0–500 μM) for 48 h, and transepithelial electrical resistance (TER) was assessed. Furthermore, BTB-associated protein expression, TNFα content, and phosphorylation and protein levels of the mitogen-activated protein kinase (MAPK) pathway were detected. An apparent decrease in mated and pregnant females per male mouse as well as litter weight was observed. Marked BTB damage was evidenced by increased red biotin fluorescence in the lumen tubular of the testes and the decrease in TER in SCs in vitro. The protein levels of claudin-11, connexin-43, N-cadherin, β-catenin, and occludin were significantly decreased in the testes and also in the SCs in vitro except for N-cadherin and β-catenin. TNFα content showed a dose-dependent increase in the testes and a dose- and time-dependent increase in the SCs, with the p-p38/p38 MAPK ratio also increasing in testes and SCs after PFOA exposure. Moreover, PFOA altered expressions of claudin-11, connexin-43, TNFα, and p-p38 MAPK were recovered 48 h after PFOA removal in the SCs. The SCs appeared to be target to PFOA, and the disruption of the BTB may be crucial to PFOA-induced reproductive dysfunction in mice.
Similar content being viewed by others
References
Aggarwal BB (2003) Signalling pathways of the TNF superfamily: a double-edged sword. Nat Rev Immunol 3(9):745–756
Alves MG, Rato L, Carvalho RA, Moreira PI, Socorro S, Oliveira PF (2013) Hormonal control of Sertoli cell metabolism regulates spermatogenesis. Cell Mol Life Sci 70(5):777–793
Apelberg BJ, Witter FR, Herbstman JB, Calafat AM, Halden RU, Needham LL, Goldman LR (2007) Cord serum concentrations of perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) in relation to weight and size at birth. Environ Health Persp 115(11):1670–1676
Booth D, Haley JD, Bruskin AM, Potten CS (2000) Transforming growth factor-B3 protects murine small intestinal crypt stem cells and animal survival after irradiation, possibly by reducing stem-cell cycling. Int J Cancer 86(1):53–59
Castiglioni S, Valsecchi S, Polesello S, Rusconi M, Melis M, Palmiotto M, Manenti A, Davoli E, Zuccato E (2014) Sources and fate of perfluorinated compounds in the aqueous environment and in drinking water of a highly urbanized and industrialized area in Italy. J Hazard Mater. doi:10.1016/j.jhazmat.2014.06.007
Coyne CB, Vanhook MK, Gambling TM, Carson JL, Boucher RC, Johnson LG (2002) Regulation of airway tight junctions by proinflammatory cytokines. Mol Biol Cell 13(9):3218–3234
Dankers AC, Roelofs MJ, Piersma AH, Sweep FC, Russel FG, van den Berg M, van Duursen MB, Masereeuw R (2013) Endocrine disruptors differentially target ATP-binding cassette transporters in the blood-testis barrier and affect Leydig cell testosterone secretion in vitro. Toxicol Sci 136(2):382–391
Fei C, McLaughlin JK, Tarone RE, Olsen J (2007) Perfluorinated chemicals and fetal growth: a study within the Danish National Birth Cohort. Environ Health Persp 115(11):1677–1682
Fei C, McLaughlin JK, Tarone RE, Olsen J (2008) Fetal growth indicators and perfluorinated chemicals: a study in the Danish National Birth Cohort. Am J Epidemiol 168(1):66–72
Feng Y, Shi Z, Fang X, Xu M, Dai J (2009) Perfluorononanoic acid induces apoptosis involving the Fas death receptor signaling pathway in rat testis. Toxicol Lett 190(2):224–230
Feng Y, Fang X, Shi Z, Xu M, Dai J (2010) Effects of PFNA exposure on expression of junction-associated molecules and secretory function in rat Sertoli cells. Reprod Toxicol 30(3):429–437
Gunther S, Fietz D, Weider K, Bergmann M, Brehm R (2013) Effects of a murine germ cell-specific knockout of Connexin 43 on Connexin expression in testis and fertility. Transgenic Res 22(3):631–641
Gunzel D, Yu AS (2013) Claudins and the modulation of tight junction permeability. Physiol Rev 93(2):525–569
He Z, Kokkinaki M, Pant D, Gallicano GI, Dym M (2009) Small RNA molecules in the regulation of spermatogenesis. Reproduction 137(6):901–911
Hellani A, Ji J, Mauduit C, Deschildre C, Tabone E, Benahmed M (2000) Developmental and hormonal regulation of the expression of oligodendrocyte-specific protein/claudin 11 in mouse testis. Endocrinology 141(8):3012–3019
Hundley SG, Sarrif AM, Kennedy GL (2006) Absorption, distribution, and excretion of ammonium perfluorooctanoate (APFO) after oral administration to various species. Drug Chem Toxicol 29(2):137–145
Joensen UN, Bossi R, Leffers H, Jensen AA, Skakkebaek NE, Jorgensen N (2009) Do perfluoroalkyl compounds impair human semen quality? Environ Health Persp 117(6):923–927
Joensen UN, Veyrand B, Antignac JP, Jensen MB, Petersen JH, Marchand P, Skakkebaek NE, Andersson AM, Le Bizec B, Jorgensen N (2014) PFOS (perfluorooctanesulfonate) in serum is negatively associated with testosterone levels, but not with semen quality, in healthy men. Hum Reprod. doi:10.1093/humrep/deu104
Lee NP, Mruk D, Lee WM, Cheng CY (2003) Is the cadherin/catenin complex a functional unit of cell–cell actin-based adherens junctions in the rat testis? Biol Reprod 68(2):489–508
Li MW, Xia W, Mruk DD, Wang CQ, Yan HH, Siu MK, Lui WY, Lee WM, Cheng CY (2006) Tumor necrosis factor alpha reversibly disrupts the blood-testis barrier and impairs Sertoli-germ cell adhesion in the seminiferous epithelium of adult rat testes. J Endocrinol 190(2):313–329
Li MW, Mruk DD, Lee WM, Cheng CY (2009) Connexin 43 and plakophilin-2 as a protein complex that regulates blood-testis barrier dynamics. Proc Natl Acad Sci USA 106(25):10213–10218
Li MW, Mruk DD, Lee WM, Cheng CY (2010) Connexin 43 is critical to maintain the homeostasis of the blood-testis barrier via its effects on tight junction reassembly. Proc Natl Acad Sci USA 107(42):17998–18003
Lie PP, Cheng CY, Mruk DD (2009) Coordinating cellular events during spermatogenesis: a biochemical model. Trends Biochem Sci 34(7):366–373
Lie PP, Cheng CY, Mruk DD (2013) Signalling pathways regulating the blood-testis barrier. Int J Biochem Cell Biol 45(3):621–625
Lindstrom AB, Strynar MJ, Libelo EL (2011) Polyfluorinated compounds: past, present, and future. Environ Sci Technol 45(19):7954–7961
Liu WY, Wang ZB, Zhang LC, Wei X, Li L (2012) Tight junction in blood-brain barrier: an overview of structure, regulation, and regulator substances. CNS Neurosci Ther 18(8):609–615
Lui WY, Wong CH, Mruk DD, Cheng CY (2003) TGF-beta3 regulates the blood-testis barrier dynamics via the p38 mitogen activated protein (MAP) kinase pathway: an in vivo study. Endocrinology 144(4):1139–1142
Luo Y, Wang X, Chen Y, Xu S, Ding G, Shi C (2013) Effects of electromagnetic radiation on morphology and TGF-beta3 expression in mouse testicular tissue. Toxicology 310:8–14
Lydka M, Bilinska B, Cheng CY, Mruk DD (2012) Tumor necrosis factor alpha-mediated restructuring of the Sertoli cell barrier in vitro involves matrix metalloprotease 9 (MMP9), membrane-bound intercellular adhesion molecule-1 (ICAM-1) and the actin cytoskeleton. Spermatogenesis 2(4):294–303
Mankertz J, Tavalali S, Schmitz H, Mankertz A, Riecken EO, Fromm M, Schulzke JD (2000) Expression from the human occludin promoter is affected by tumor necrosis factor alpha and interferon gamma. J Cell Sci 113(Pt 11):2085–2090
Meng J, Holdcraft RW, Shima JE, Griswold MD, Braun RE (2005) Androgens regulate the permeability of the blood-testis barrier. Proc Natl Acad Sci USA 102(46):16696–16700
Mocellin S, Rossi CR, Pilati P, Nitti D (2005) Tumor necrosis factor, cancer and anticancer therapy. Cytokine Growth Factor Rev 16(1):35–53
Mruk DD, Cheng CY (2011) An in vitro system to study Sertoli cell blood-testis barrier dynamics. Methods Mol Biol 763:237–252
Olsen GW, Burris JM, Burlew MM, Mandel JH (2000) Plasma cholecystokinin and hepatic enzymes, cholesterol and lipoproteins in ammonium perfluorooctanoate production workers. Drug Chem Toxicol 23(4):603–620
Pico Y, Farre M, Llorca M, Barcelo D (2011) Perfluorinated compounds in food: a global perspective. Crit Rev Food Sci Nutr 51(7):605–625
Post GB, Cohn PD, Cooper KR (2012) Perfluorooctanoic acid (PFOA), an emerging drinking water contaminant: a critical review of recent literature. Environ Res 116:93–117
Qiu L, Zhang X, Zhang X, Zhang Y, Gu J, Chen M, Zhang Z, Wang X, Wang SL (2013) Sertoli cell is a potential target for perfluorooctane sulfonate-induced reproductive dysfunction in male mice. Toxicol Sci 135(1):229–240
Runkle EA, Mu D (2013) Tight junction proteins: from barrier to tumorigenesis. Cancer Lett 337(1):41–48
Sabio G, Davis RJ (2014) TNF and MAP kinase signalling pathways. Semin Immunol 26(3):237–245
Saunders PT (2003) Germ cell-somatic cell interactions during spermatogenesis. Reprod Suppl 61:91–101
Shaw SK, Perkins BN, Lim YC, Liu Y, Nusrat A, Schnell FJ, Parkos CA, Luscinskas FW (2001) Reduced expression of junctional adhesion molecule and platelet/endothelial cell adhesion molecule-1 (CD31) at human vascular endothelial junctions by cytokines tumor necrosis factor-alpha plus interferon-gamma does not reduce leukocyte transmigration under flow. Am J Pathol 159(6):2281–2291
Shi Z, Hou J, Guo X, Zhang H, Yang F, Dai J (2013) Testicular phosphoproteome in perfluorododecanoic acid-exposed rats. Toxicol Lett 221(2):91–101
Siu ER, Mruk DD, Porto CS, Cheng CY (2009) Cadmium-induced testicular injury. Toxicol Appl Pharmacol 238(3):240–249
Steenland K, Jin C, MacNeil J, Lally C, Ducatman A, Vieira V, Fletcher T (2009) Predictors of PFOA levels in a community surrounding a chemical plant. Environ Health Perspect 117(7):1083–1088
Steinberger A, Klinefelter G (1993) Sensitivity of Sertoli and Leydig cells to xenobiotics in in vitro models. Reprod Toxicol 7(Suppl 1):23–37
Tsukita S, Furuse M, Itoh M (2001) Multifunctional strands in tight junctions. Nat Rev Mol Cell Biol 2(4):285–293
Vested A, Ramlau-Hansen CH, Olsen SF, Bonde JP, Kristensen SL, Halldorsson TI, Becher G, Haug LS, Ernst EH, Toft G (2013) Associations of in utero exposure to perfluorinated alkyl acids with human semen quality and reproductive hormones in adult men. Environ Health Perspect 121(4):453–458
Vestergren RC, Cousins IT (2009) Tracking the pathways of human exposure to perfluorocarboxylates. Environ Sci Technol 43(15):5565–5575
Wan HT, Mruk DD, Wong CK, Cheng CY (2014) Perfluorooctanesulfonate (PFOS) perturbs male rat Sertoli cell blood-testis barrier function by affecting F-actin organization via p-FAK-Tyr(407): an in vitro study. Endocrinology 155(1):249–262
White SS, Fenton SE, Hines EP (2011) Endocrine disrupting properties of perfluorooctanoic acid. J Steroid Biochem Mol Biol 127(1–2):16–26
Xia W, Mruk DD, Lee WM, Cheng CY (2005) Cytokines and junction restructuring during spermatogenesis—a lesson to learn from the testis. Cytokine Growth Factor Rev 16(4–5):469–493
Xia W, Wong EW, Mruk DD, Cheng CY (2009) TGF-beta3 and TNFalpha perturb blood-testis barrier (BTB) dynamics by accelerating the clathrin-mediated endocytosis of integral membrane proteins: a new concept of BTB regulation during spermatogenesis. Dev Biol 327(1):48–61
Xiao X, Wong EW, Lie PP, Mruk DD, Wong CK, Cheng CY (2014) Cytokines, polarity proteins, and endosomal protein trafficking and signaling-the Sertoli cell blood-testis barrier system in vitro as a study model. Methods Enzymol 534:181–194
Yan S, Wang J, Dai J (2014) Activation of sterol regulatory element-binding proteins in mice exposed to perfluorooctanoic acid for 28 days. Arch Toxicol. doi:10.1007/s00204-014-1322-7
Zhang J, Wong CH, Xia W, Mruk DD, Lee NP, Lee WM, Cheng CY (2005) Regulation of Sertoli-germ cell adherens junction dynamics via changes in protein-protein interactions of the N-cadherin-beta-catenin protein complex which are possibly mediated by c-Src and myotubularin-related protein 2: an in vivo study using an androgen suppression model. Endocrinology 146(3):1268–1284
Zhang YH, Lin L, Liu ZW, Jiang XZ, Chen BH (2008) Disruption effects of monophthalate exposures on inter-Sertoli tight junction in a two-compartment culture model. Environ Toxicol 23(3):302–308
Zhang H, Lu Y, Luo B, Yan S, Guo X, Dai J (2014) Proteomic analysis of mouse testis reveals perfluorooctanoic acid-induced reproductive dysfunction via direct disturbance of testicular steroidogenic machinery. J Proteome Res 13(7):3370–3385
Zhao B, Chu Y, Hardy DO, Li XK, Ge RS (2010) Inhibition of 3beta- and 17beta-hydroxysteroid dehydrogenase activities in rat Leydig cells by perfluorooctane acid. J Steroid Biochem Mol Biol 118(1–2):13–17
Acknowledgments
This work was supported by the National Basic Research Program of China (973; Grant: 2013CB945204) and the National Natural Science Foundation of China (Grants: 31320103915 and 31025006).
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Lu, Y., Luo, B., Li, J. et al. Perfluorooctanoic acid disrupts the blood–testis barrier and activates the TNFα/p38 MAPK signaling pathway in vivo and in vitro. Arch Toxicol 90, 971–983 (2016). https://doi.org/10.1007/s00204-015-1492-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-015-1492-y