Skip to main content
Log in

Effects of clothianidin exposure on sperm quality, testicular apoptosis and fatty acid composition in developing male rats

  • Original Research
  • Published:
Cell Biology and Toxicology Aims and scope Submit manuscript

Abstract

Clothianidin (CTD) is one of the latest members of the synthetic organic insecticides, the neonicotinoids. In the present study, it was aimed to investigate if daily oral administration of CTD at low doses for 90 days has any deleterious effects on reproductive functions of developing male rats. Animals were randomly divided into four groups of six rats each, assigned as control rats, or rats treated with 2 (CTD-2), 8 (CTD-8) or 32 (CTD-32) mg CTD/kg body weight by oral gavage. The significant decreases of the absolute weights of right cauda epididymis and seminal vesicles, and body weight were detected in the animals exposed to CTD administration at 32 mg/kgBW/day. Epididymal sperm concentration decreased significantly in CTD-32 group and the abnormal sperm rates increased in CTD-8 and CTD-32 groups when compared to control group. The testosterone level was significantly decreased in CTD-32 group when compared to control group. The administration of all CTD doses resulted in a significant decrease in the level of GSH. The number of TUNEL-positive cells significantly increased in the germinal epithelium of testis of rats exposed to CTD at 32 mg/kgBW/day. In groups CTD-8 and CTD-32, only docosapentaenoic, arachidonic, palmitic and palmitoleic acids were significantly elevated when compared to control. The ratios of 20:4/18:2 and 18:1n−9/18:0 were decreased when rats exposed to CTD. Sperm DNA fragmentation was observed in CTD-32 group, but not CTD-2 and CTD-8. It is concluded that low doses of CTD exposure during critical stages of sexual maturation had moderate detrimental effects on reproductive organ system and more severe effects are likely to be observed at higher dose levels. In addition, the reproductive system may be more sensitive to exposure of CTD even earlier in development (prenatal and early postnatal), and therefore it could be expected that more severe effects could also be observed at the NOAEL dose levels, if dosing had occurred in utero or early postnatal.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Agarwal A, Makker K, Sharma R. Clinical relevance of oxidative stress in male factor infertility: an update. Am J Reprod Immunol. 2008;59:2–11.

    Article  PubMed  CAS  Google Scholar 

  • Akerboom TP, Sies H. Assay of glutathione, glutathione disulfide, and glutathione mixed disulfides in biological samples. Methods Enzymol. 1981;77:373–82.

    Article  PubMed  CAS  Google Scholar 

  • Akingbemi BT, Ge R, Hardy MP. Leydig cells. In: Knobil E, Neill JD, editors. Encyclopedia of reproduction. San Diego: Academic; 1999. p. 1021–33.

    Google Scholar 

  • Akingbemi BT, Ge RS, Klinefelter GR, Gunsalus GL, Hardy MP. A metabolite of methoxychlor, 2,2-bis(p-hydroxyphenyl)-1,1, 1-trichloroethane, reduces testosterone biosynthesis in rat leydig cells through suppression of steady-state messenger ribonucleic acid levels of the cholesterol side-chain cleavage enzyme. Biol Reprod. 2000;62:571–8.

    Article  PubMed  CAS  Google Scholar 

  • Altuğ ME, Serarslan Y, Bal R, Kontaş T, Ekici F, Melek IM, Aslan H, Duman T. Caffeic acid phenethyl ester protects rabbit brains against permanent focal ischemia by antioxidant action: a biochemical and planimetric study. Brain Res. 2008;1201:135–42.

    Article  PubMed  Google Scholar 

  • Ayala S, Brenner RR, Dumm CG. Effect of polyunsaturated fatty acids of the alpha-linolenic series on the development of rat testicles. Lipids. 1977;12:1017–24.

    Article  PubMed  CAS  Google Scholar 

  • Bal R, Oertel D. Voltage-activated calcium currents in octopus cells of the mouse cochlear nucleus. J Assoc Res Otolaryngol. 2007;8:509–21.

    Article  PubMed  Google Scholar 

  • Bal R, Erdogan S, Theophilidis G, Baydas G, Naziroglu M. Assessing the effects of the neonicotinoid insecticide imidacloprid in the cholinergic synapses of the stellate cells of the mouse cochlear nucleus using whole-cell patch-clamp recording. Neurotoxicology. 2010;31:113–20.

    Article  PubMed  CAS  Google Scholar 

  • Bal R, Türk G, Tuzcu M, Yılmaz Ö, Özercan İ, Kuloğlu T, Gür S, Nedzvetsky VS, Tykhomyrov AA, Andrievsky GV, Baydaş G, Nazıroğlu M. Protective effects of nanostructures of hydrated C(60) fullerene on reproductive function in streptozotocin-diabetic male rats. Toxicology. 2011;282:69–81.

    Article  PubMed  CAS  Google Scholar 

  • Barkley MS, Goldman BD. A quantitative study of serum testosterone, sex accessory organ growth, and the development of intermale aggression in the mouse. Horm Behav. 1977;8:208–18.

    Article  PubMed  CAS  Google Scholar 

  • Bhardwaj S, Srivastava MK, Kapoor U, Srivastava LP. A 90 days oral toxicity of imidacloprid in female rats: morphological, biochemical and histopathological evaluations. Food Chem Toxicol. 2010;48:1185–90.

    Article  PubMed  CAS  Google Scholar 

  • Bian Q, Xu LC, Wang SL, Xia YK, Tan LF, Chen JF, Song L, Chang HC, Wang XR. Study on the relation between occupational fenvalerate exposure and spermatozoa DNA damage of pesticide factory workers. Occup Environ Med. 2004;61:999–1005.

    Article  PubMed  CAS  Google Scholar 

  • Bourre JM, Dumont OL, Clement ME, Durand GA. Endogenous synthesis cannot compensate for absence of dietary oleic acid in rats. J Nutr. 1997;127:488–93.

    PubMed  CAS  Google Scholar 

  • Bray C, Son J-H, Kumar P, Meizel S. Mice deficient in CHRNA7, a subunit of the nicotinic acetylcholine receptor, produce sperm with impaired motility. Biol Reprod. 2005;73:807–14.

    Article  PubMed  CAS  Google Scholar 

  • Catala A. The ability of melatonin to counteract lipid peroxidation in biological membranes. Curr Mol Med. 2007;7:638–49.

    Article  PubMed  CAS  Google Scholar 

  • Christie WW. Gas chromatography and lipids. Glasgow: The Oil; 1992.

    Google Scholar 

  • Davis JT, Coniglio JG. The biosynthesis of docosapentaenoic and other fatty acids by rat testes. J Biol Chem. 1966;241:610–2.

    PubMed  CAS  Google Scholar 

  • de las Heras A, Schoch AM, Fischer A. Comparison of methods for determining malondialdehyde in dry sausage by HPLC and the classic TBA test. Eur Food Res Technol. 2003;217:180–4.

    Article  Google Scholar 

  • El-Gendy KS, Aly NM, Mahmoud FH, Kenawy A, El-Sebae AKH. The role of vitamin C as antioxidant in protection of oxidative stress induced by imidacloprid. Food Chem Toxicol. 2010;48:215–21.

    Article  PubMed  CAS  Google Scholar 

  • El-Gohary M, Awara WM, Nassar S, Hawas S. Deltamethrin-induced testicular apoptosis in rats: the protective effect of nitric oxide synthase inhibitor. Toxicology. 1999;132:1–8.

    Article  PubMed  CAS  Google Scholar 

  • Ellman GL. Tissue sulfhydryl groups. Arch Biochem Biophys. 1959;82:70–7.

    Article  PubMed  CAS  Google Scholar 

  • El-Sheekh MM, Kotkat HM, Hammouda OHE. Effect of atrazine herbicide on growth, photosynthesis, protein synthesis, and fatty acid composition in the unicellular green alga Chlorella kessleri. Ecotoxicol Environ Saf. 1994;29:349–58.

    Article  PubMed  CAS  Google Scholar 

  • Ewing LL, Keeney DS. Leydig cells: structure and function. In: Desjardins C, Ewing LL, editors. Cell and molecular biology of the testis. New York: Oxford University Press; 1993. pp. 137–65.

  • Federal Register. Clothianidin; pesticide tolerances. Authenticated US Govern Inform. 2003;68:32390–400.

    Google Scholar 

  • Fernandes GSA, Arena AC, Fernadez CDB, Mercadante A, Barbisan LF, Kempinas WG. Reproductive effects in male rats exposed to diuron. Reprod Toxicol. 2007;23:106–12.

    Article  PubMed  CAS  Google Scholar 

  • Flora A, Schulz R, Benfante R, Battaglioli E, Terzano S, Clementi F, Fornasari D. Transcriptional regulation of the human α5 nicotinic receptor subunit gene in neuronal and non-neuronal tissues. Eur J Pharmacol. 2000;393:85–95.

    Article  PubMed  CAS  Google Scholar 

  • Gutteridge JM, Quinlan GJ, Yamamoto Y. Hypothesis: are fatty acid patterns characteristic of essential fatty acid deficiency indicative of oxidative stress? Free Radic Res. 1998;28:109–14.

    Article  PubMed  CAS  Google Scholar 

  • Hammer CT, Wills ED. The role of lipid components of the diet in the regulation of the fatty acid composition of the rat liver endoplasmic reticulum and lipid peroxidation. Biochem J. 1978;174:585–93.

    PubMed  CAS  Google Scholar 

  • Hara A, Radin NS. Lipid extraction of tissues with a low-toxicity solvent. Anal Biochem. 1978;90:420–6.

    Article  PubMed  CAS  Google Scholar 

  • Hurtado de Catalfo GE, de Gomez Dumm IN. Influence of testosterone on polyunsaturated fatty acid biosynthesis in Sertoli cells in culture. Cell Biochem Funct. 2005;23:175–80.

    Article  PubMed  CAS  Google Scholar 

  • Johnson AD. The testis. New York: Academic; 1970.

    Google Scholar 

  • Kaur B, Sandhu HS, Kaur R. Toxic effects of subacute oral exposure of imidacloprid on biochemical parameters in crossbred cow calves. Toxicol Int. 2006;13:43–7.

    CAS  Google Scholar 

  • Kawai Y, Hata T, Suzuki O, Matsuda J. The relationship between sperm morphology and in vitro fertilization ability in mice. J Reprod Dev. 2006;52:561–8.

    Article  PubMed  Google Scholar 

  • Klinefelter GR, Hess RA. Toxicity of the excurrent ducts and accessory organs in the male. In: Korach KS, editor. Reproductive and developmental toxicology. New York: Marcel Dekker; 1998. p. 553–91.

    Google Scholar 

  • Lenzi A, Picardo M, Gandini L, Dondero F. Lipids of the sperm plasma membrane: from polyunsaturated fatty acids considered as markers of sperm function to possible scavenger therapy. Hum Reprod Update. 1996;2:246–56.

    Article  PubMed  CAS  Google Scholar 

  • Li P, Ann J, Akk G. Activation and modulation of human α4β2 nicotinic acetylcholine receptors by the neonicotinoids clothianidin and imidacloprid. J Neurosci Res. 2011;89:1295–301.

    Article  PubMed  CAS  Google Scholar 

  • Lopez-Cervantes J, Sanchez-Machado DI, Rios-Vazquez NJ. High-performance liquid chromatography method for the simultaneous quantification of retinol, alpha-tocopherol, and cholesterol in shrimp waste hydrolysate. J Chromatogr A. 2006;1105:135–9.

    Article  PubMed  CAS  Google Scholar 

  • Mallick C, Mandal S, Barik B, Bhattacharya A, Ghosh D. Protection of testicular dysfunctions by MTEC, a formulated herbal drug, in streptozotocin induced diabetic rat. Biol Pharm Bull. 2007;30:84–90.

    Article  PubMed  CAS  Google Scholar 

  • Meikle AW, Benson SJ, Liu XH, Boam WD, Stringham JD. Nonesterified fatty acids modulate steroidogenesis in mouse Leydig cells. Am J Physiol. 1989;257:E937–42.

    PubMed  CAS  Google Scholar 

  • Meikle AW, Cardoso de Sousa JC, Hanzalova J, Murray DK. Oleic acid inhibits cholesteryl esterase and cholesterol utilization for testosterone synthesis in mouse Leydig cells. Metabolism. 1996;45:293–9.

    Article  PubMed  CAS  Google Scholar 

  • Mohany M, Badr G, Refaat I, El-Feki M. Immunological and histological effects of exposure to imidacloprid insecticide in male albino rats. Afr J Pharm Pharmacol. 2011;5:2106–14.

    CAS  Google Scholar 

  • Najafi G, Razi M, Hoshyar A, Shahmohamadloo S, Feyzi S. The effect of chronic exposure with imidacloprid insecticide on fertility in mature male rats. Int J Fertil Steril. 2010;4:9–16.

    Google Scholar 

  • Niehaus WGJ, Samuelsson B. Formation of malonaldehyde from phospholipid arachidonate during microsomal lipid peroxidation. Eur J Biochem. 1968;6:126–30.

    Article  PubMed  CAS  Google Scholar 

  • Ngoula F, Watcho P, Dongmo MC, Kenfack A, Kamtchouing P, Tchoumboue J. Effects of pirimiphos-methyl (an organophosphate insecticide) on the fertility of adult male rats. Afr Health Sci. 2007;7:3–9.

    PubMed  Google Scholar 

  • Oertel D, Wright S, Cao XJ, Ferragamo M, Bal R. The multiple functions of T stellate/multipolar/chopper cells in the ventral cochlear nucleus. Hear Res. 2011;276:61–9.

    Article  PubMed  Google Scholar 

  • Peltola V, Huhtaniemi I, Ahotupa M. Antioxidant enzyme activity in the maturing rat testis. J Androl. 1992;13:450–5.

    PubMed  CAS  Google Scholar 

  • Pompeia C, Freitas JJ, Kim JS, Zyngier SB, Curi R. Arachidonic acid cytotoxicity in leukocytes: implications of oxidative stress and eicosanoid synthesis. Biol Cell. 2002;94:251–65.

    Article  PubMed  CAS  Google Scholar 

  • Samanta L, Chainy GB. Comparison of hexachlorocyclohexane-induced oxidative stress in the testis of immature and adult rats. Comp Biochem Physiol C Pharmacol Toxicol Endocrinol. 1997;118:319–27.

    Article  PubMed  CAS  Google Scholar 

  • Samanta L, Roy A, Chainy GB. Changes in rat testicular antioxidant defence profile as a function of age and its impairment by hexachlorocyclohexane during critical stages of maturation. Andrologia. 1999;31:83–90.

    PubMed  CAS  Google Scholar 

  • Sanchez-Machado DI, Lopez-Hernandez J, Paseiro-Losada P, Lopez-Cervantes J. An HPLC method for the quantification of sterols in edible seaweeds. Biomed Chromatogr. 2004;18:183–90.

    Google Scholar 

  • Saradha B, Vaithinathan S, Mathur PP. Lindane induces testicular apoptosis in adult Wistar rats through the involvement of Fas-FasL and mitochondria-dependent pathways. Toxicology. 2009;255:131–9.

    Article  PubMed  CAS  Google Scholar 

  • Sumitomo Chemical Takeda Agro Company, Ltd. Technical information: summary of toxicity studies with clothianidin (in Japanese). J Pest Sci. 2003;28:167–73.

    Google Scholar 

  • Tanaka T. Effects of maternal clothianidin exposure on behavioral development in F1 generation mice. Toxicol Ind Health. 2012. doi:10.1177/0748233711422726.

  • Tomizawa M, Casida JE. Selective toxicity of neonicotinoids attributable to specificity of insect and mammalian nicotinic receptors. Annu Rev Entomol. 2003;48:339–64.

    Article  PubMed  CAS  Google Scholar 

  • Tomizawa M, Casida JE. Neonicotinoid insecticide toxicology: mechanisms of selective action. Annu Rev Pharmacol Toxicol. 2005;45:247–68.

    Article  PubMed  CAS  Google Scholar 

  • Tomizawa M, Lee DL, Casida JE. Neonicotinoid insecticides: molecular features conferring selectivity for insect versus mammalian nicotinic receptors. J Agric Food Chem. 2000;48:6016–24.

    Article  PubMed  CAS  Google Scholar 

  • Türk G, Ateşşahin A, Sönmez M, Çeribaşı AO, Yüce A. Improvement of cisplatin-induced injuries to sperm quality, the oxidant-antioxidant system, and the histologic structure of the rat testis by ellagic acid. Fertil Steril. 2008;89:1474–81.

    Article  PubMed  Google Scholar 

  • Türk G, Çeribaşı AO, Sakin F, Sönmez M, Ateşşahin A. Antiperoxidative and anti-apoptotic effects of lycopene and ellagic acid on cyclophosphamide-induced testicular lipid peroxidation and apoptosis. Reprod Fertil Dev. 2010;22:587–96.

    Article  PubMed  Google Scholar 

  • Vaithinathan S, Saradha B, Mathur PP. Methoxychlor induces apoptosis via mitochondria- and FasL-mediated pathways in adult rat testis. Chem Biol Interact. 2010;185:110–8.

    Article  PubMed  CAS  Google Scholar 

  • Vento R, D'Alessandro N, Giuliano M, Lauricella M, Carabillo M, Tesoriere G. Induction of apoptosis by arachidonic acid in human retinoblastoma Y79 cells: involvement of oxidative stress. Exp Eye Res. 2000;70:503–17.

    Article  PubMed  CAS  Google Scholar 

  • Walsh LP, McCormick C, Martin C, Stocco DM. Roundup inhibits steroidogenesis by disrupting steroidogenic acute regulatory (StAR) protein expression. Environ Health Perspect. 2000;108:769–76.

    Article  PubMed  CAS  Google Scholar 

  • Wang GM, Ge RS, Latif SA, Morris DJ, Hardy MP. Expression of 11beta-hydroxylase in rat Leydig cells. Endocrinology. 2002;143:621–6.

    Article  PubMed  CAS  Google Scholar 

  • Wang JY, Shum AY, Ho YJ, Wang JY. Oxidative neurotoxicity in rat cerebral cortex neurons: synergistic effects of H2O2 and NO on apoptosis involving activation of p38 mitogen-activated protein kinase and caspase-3. J Neurosci Res. 2003;72:508–19.

    Article  PubMed  CAS  Google Scholar 

  • Yamamoto Y, Shimamoto K, Sofikitis N, Miyagawa I. Effects of hypercholesterolaemia on Leydig and Sertoli cell secretory function and the overall sperm fertilizing capacity in the rabbit. Hum Reprod. 1999;14:1516–21.

    Article  PubMed  CAS  Google Scholar 

  • Yener Z, Celik I, Ilhan F, Bal R. Effects of urtica dioica L. seed on lipid peroxidation, antioxidants and liver pathology in aflatoxin-induced tissue injury in rats. Food Chem Toxicol. 2009;47:418–24.

    Article  PubMed  CAS  Google Scholar 

  • Yokota T, Mikata K, Nagasaki H, Ohta K. Absorption, tissue distribution, excretion, and metabolism of clothianidin in rats. J Agric Food Chem. 2003;51:7066–72.

    Article  PubMed  CAS  Google Scholar 

  • Zhang J-J, Wang Y, Xıang H-Y, Li M-X, Li W-H, Ma K-G, Wang X-Z, Zhang J-H. Oxidative stress: role in acetamiprid-induced impairment of the male mice reproductive system. Agr Sci China. 2011;10:786–96.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Gaffari Türk.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bal, R., Türk, G., Yılmaz, Ö. et al. Effects of clothianidin exposure on sperm quality, testicular apoptosis and fatty acid composition in developing male rats. Cell Biol Toxicol 28, 187–200 (2012). https://doi.org/10.1007/s10565-012-9215-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10565-012-9215-0

Keywords

Navigation