Neurotoxicity Research

, Volume 36, Issue 4, pp 700–711 | Cite as

Chlorpyrifos Exposure Induces Parkinsonian Symptoms and Associated Bone Loss in Adult Swiss Albino Mice

  • Shaheen Jafri Ali
  • Govindraj Ellur
  • Kalpana Patel
  • Kunal SharanEmail author
Original Article


Prenatal and early life exposure of chlorpyrifos (CPF), a widely used pesticide, is known to cause neuronal deficits and Parkinson’s disease (PD). However, data about the effect of its exposure at adult stages on PD-like symptoms and associated bone loss is scanty. In the present study, we investigated the impact of CPF on the behavioral alterations seen in PD using adult Swiss albino mice. PD is often associated with bone loss. Hence, skeletal changes were also evaluated using micro-computed tomography and histology. MPTP was used as a positive control. Cell culture studies using MC3T3E-1, SHSY5Y, and primary osteoclast cultures were done to understand the cellular mechanism for the behavioral and skeletal changes. Our results showed that CPF treatment leads to PD-like symptoms due to the loss of dopaminergic neurons. Moreover, CPF has a deleterious effect on the trabecular bone through both indirect changes in circulating factors and direct stimulation of multinucleate osteoclast cell formation. The impact on the bone mass was even stronger than MPTP. In conclusion, this is the first report demonstrating that CPF induces parkinsonian features in adult Swiss albino mice and it is accompanied by loss of trabecular bone.


Chlorpyrifos Parkinson’s disease Osteoporosis Dopamine Osteoclast 



This study was supported by CSIR-Central Food Technological Research Institute, Mysore, India, and SERB N-PDF grant. Funding from the Science and Engineering Research Board (SERB), Government of India (K.S.), and the Department of Biotechnology, Government of India (K.S.), is acknowledged. Research fellowship grants from the SERB (S.J.A.), Department of Science and Technology (G.E.), and Council of Scientific and Industrial Research (K.P), Government of India, are also acknowledged. We thank Dr. Naibedya Chattopadhyay (CSIR-Central Drug Research Institute, Lucknow, India) for the μ-CT facility.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

12640_2019_92_MOESM1_ESM.docx (12 kb)
Supplementary table 1 (DOCX 12 kb)


  1. Aldridge JE, Meyer A, Seidler FJ, Slotkin TA (2005) Alterations in central nervous system serotonergic and dopaminergic synaptic activity in adulthood after prenatal or neonatal chlorpyrifos exposure. Environ Health Perspect 113:1027–1031. CrossRefPubMedPubMedCentralGoogle Scholar
  2. Ali SJ, Rajini PS (2016) Effect of monocrotophos, an organophosphorus insecticide, on the striatal dopaminergic system in a mouse model of Parkinson’s disease. Toxicol Ind Health 32:1153–1165. CrossRefPubMedGoogle Scholar
  3. Ali SJ, Ellur G, Khan MT, Sharan K (2019) Bone loss in MPTP mouse model of Parkinson’s disease is triggered by decreased osteoblastogenesis and increased osteoclastogenesis. Toxicol Appl Pharmacol 363:154–163. CrossRefPubMedGoogle Scholar
  4. Bachagol D, Joseph GS, Ellur G, Patel K, Aruna P et al (2018) Stimulation of liver IGF-1 expression promotes peak bone mass achievement in growing rats: a study with pomegranate seed oil. J Nutr Biochem 52:18–26. CrossRefPubMedGoogle Scholar
  5. Basaure P, Guardia-Escote L, Cabre M, Peris-Sampedro F, Sanchez-Santed F et al (2018) Postnatal chlorpyrifos exposure and apolipoprotein E (APOE) genotype differentially affect cholinergic expression and developmental parameters in transgenic mice. Food Chem Toxicol 118:42–52. CrossRefPubMedGoogle Scholar
  6. Beitz JM (2014) Parkinson’s disease: a review. Front Biosci (Schol Ed) 6:65–74CrossRefGoogle Scholar
  7. Cecchini MP, Federico A, Zanini A, Mantovani E, Masala C et al. (2019) Olfaction and taste in Parkinson’s disease: the association with mild cognitive impairment and the single cognitive domain dysfunction. CrossRefGoogle Scholar
  8. Cheng WW, Zhu Q, Zhang HY (2019) Mineral nutrition and the risk of chronic diseases: a Mendelian randomization study. Nutrients 11. CrossRefGoogle Scholar
  9. Choi SM, Kim BC, Jung HJ, Yoon GJ, Kang KW et al (2017) The association of musculoskeletal pain with bone mineral density in patients with Parkinson’s disease. Eur Neurol 77:123–129. CrossRefPubMedGoogle Scholar
  10. Clevelan CB, Oliver GR, Chen B, Mattsson J (2001) Risk assessment under FQPA: case study with chlorpyrifos. Neurotoxicology 22:699–706CrossRefGoogle Scholar
  11. Cummings SR, Eastell R (2016) Risk and prevention of fracture in patients with major medical illnesses: a mini-review. J Bone Miner Res Off J Am Soc Bone Miner Res 31:2069–2072. CrossRefGoogle Scholar
  12. Dam K, Seidler FJ, Slotkin TA (2000) Chlorpyrifos exposure during a critical neonatal period elicits gender-selective deficits in the development of coordination skills and locomotor activity. Brain Res Dev Brain Res 121:179–187CrossRefGoogle Scholar
  13. Deacon RM (2013) Measuring the strength of mice. J Vis Exp.
  14. Dobson R, Yarnall A, Noyce AJ, Giovannoni G (2013) Bone health in chronic neurological diseases: a focus on multiple sclerosis and parkinsonian syndromes. Pract Neurol 13:70–79. CrossRefPubMedGoogle Scholar
  15. Eaton DL, Daroff RB, Autrup H, Bridges J, Buffler P et al (2008) Review of the toxicology of chlorpyrifos with an emphasis on human exposure and neurodevelopment. Crit Rev Toxicol 38(Suppl 2):1–125. CrossRefPubMedGoogle Scholar
  16. Ellman GL, Courtney KD, Andres V Jr, Feather-Stone RM (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95CrossRefGoogle Scholar
  17. El-Sebae AH, Ahmed NS, Soliman SA (1978) Effect of pre-exposure on acute toxicity of organophosphorus insecticides to white mice. J Environ Sci Health B 13:11–24. CrossRefPubMedGoogle Scholar
  18. Engel SM, Wetmur J, Chen J, Zhu C, Barr DB et al (2011) Prenatal exposure to organophosphates, paraoxonase 1, and cognitive development in childhood. Environ Health Perspect 119:1182–1188. CrossRefPubMedPubMedCentralGoogle Scholar
  19. Fukuda T, Takeda S (2015) Frontiers in live bone imaging researches. Functional cross talk between bone and nervous system. Clin Calcium 25:891–898PubMedGoogle Scholar
  20. Grandjean P, Landrigan PJ (2014) Neurobehavioural effects of developmental toxicity. Lancet Neurol 13:330–338. CrossRefPubMedPubMedCentralGoogle Scholar
  21. Gruntenko NE, Karpova EK, Alekseev AA, Chentsova NA, Saprykina ZV et al (2005) Effects of dopamine on juvenile hormone metabolism and fitness in Drosophila virilis. J Insect Physiol 51:959–968. CrossRefPubMedGoogle Scholar
  22. Huang S, Li Z, Liu Y, Gao D, Zhang X et al (2019) Neural regulation of bone remodeling: identifying novel neural molecules and pathways between brain and bone. J Cell Physiol 234:5466–5477. CrossRefPubMedGoogle Scholar
  23. Jackson-Lewis V, Przedborski S (2007) Protocol for the MPTP mouse model of Parkinson’s disease. Nat Protoc 2:141–151. CrossRefPubMedGoogle Scholar
  24. Jiang P (2019) Parkinson’s disease is associated with dysregulations of a dopamine-modulated gene network relevant to sleep and affective neurobehaviors in the striatum. Sci Rep 18:4808. CrossRefGoogle Scholar
  25. Kalia LV, Lang AE (2015) Parkinson’s disease. Lancet (London, England) 386:896–912. CrossRefGoogle Scholar
  26. Khan K, Singh A, Mittal M, Sharan K, Singh N et al (2012) [6]-Gingerol induces bone loss in ovary intact adult mice and augments osteoclast function via the transient receptor potential vanilloid 1 channel. Mol Nutr Food Res 56:1860–1873. CrossRefPubMedGoogle Scholar
  27. Khan MP, Mishra JS, Sharan K, Yadav M, Singh AK et al (2013) A novel flavonoid C-glucoside from Ulmus wallichiana preserves bone mineral density, microarchitecture and biomechanical properties in the presence of glucocorticoid by promoting osteoblast survival: a comparative study with human parathyroid hormone. Phytomedicine 20:1256–1266. CrossRefPubMedGoogle Scholar
  28. Kim HH, Lim YW, Yang JY, Shin DC, Ham HS et al (2013) Health risk assessment of exposure to chlorpyrifos and dichlorvos in children at childcare facilities. Sci Total Environ 444:441–450. CrossRefPubMedGoogle Scholar
  29. Konradsen F (2007) Acute pesticide poisoning—a global public health problem. Dan Med Bull 54:58–59PubMedGoogle Scholar
  30. Lewis KE, Sharan K, Takumi T, Yadav VK (2017) Skeletal site-specific changes in bone mass in a genetic mouse model for human 15q11-13 duplication seen in autism. Sci Rep 7:9902. CrossRefPubMedPubMedCentralGoogle Scholar
  31. Lozowicka B (2015) Health risk for children and adults consuming apples with pesticide residue. Sci Total Environ 502:184–198. CrossRefPubMedGoogle Scholar
  32. Metta V, Sanchez TC, Padmakumar C (2017) Osteoporosis: a hidden nonmotor face of Parkinson’s disease. Int Rev Neurobiol 134:877–890. CrossRefPubMedGoogle Scholar
  33. Mojsak P, Lozowicka B, Kaczynski P (2018) Estimating acute and chronic exposure of children and adults to chlorpyrifos in fruit and vegetables based on the new, lower toxicology data. Ecotoxicol Environ Saf 159:182–189. CrossRefPubMedGoogle Scholar
  34. Opara J, Malecki A, Malecka E, Socha T (2017) Motor assessment in Parkinson’s disease. Ann Agric Environ Med 24:411–415. CrossRefPubMedGoogle Scholar
  35. Presgraves SP, Ahmed T, Borwege S, Joyce JN (2004) Terminally differentiated SH-SY5Y cells provide a model system for studying neuroprotective effects of dopamine agonists. Neurotox Res 5:579–598CrossRefGoogle Scholar
  36. Rauh VA (2018) Polluting developing brains—EPA failure on chlorpyrifos. N Engl J Med 378:1171–1174. CrossRefPubMedGoogle Scholar
  37. Roman-Garcia P, Quiros-Gonzalez I, Mottram L, Lieben L, Sharan K et al (2014) Vitamin B(1)(2)-dependent taurine synthesis regulates growth and bone mass. J Clin Invest 124:2988–3002. CrossRefPubMedPubMedCentralGoogle Scholar
  38. Sanberg PR, Bunsey MD, Giordano M, Norman AB (1988) The catalepsy test: its ups and downs. Behav Neurosci 102:748–759CrossRefGoogle Scholar
  39. Schneider JL, Fink HA, Ewing SK, Ensrud KE, Cummings SR (2008) The association of Parkinson’s disease with bone mineral density and fracture in older women. Osteoporos Int 19:1093–1097. CrossRefPubMedGoogle Scholar
  40. Schneider RB, Iourinets J, Richard IH (2017) Parkinson’s disease psychosis: presentation, diagnosis and management. Neurodegener Dis Manag 7:365–376. CrossRefPubMedGoogle Scholar
  41. Sharan K, Yadav VK (2014) Hypothalamic control of bone metabolism. Best Pract Res Clin Endocrinol Metab 28:713–723. CrossRefPubMedGoogle Scholar
  42. Sharan K, Mishra JS, Swarnkar G, Siddiqui JA, Khan K et al (2011) A novel quercetin analogue from a medicinal plant promotes peak bone mass achievement and bone healing after injury and exerts an anabolic effect on osteoporotic bone: the role of aryl hydrocarbon receptor as a mediator of osteogenic action. J Bone Miner Res 26:2096–2111. CrossRefPubMedGoogle Scholar
  43. Sharan K, Lewis K, Furukawa T, Yadav VK (2017) Regulation of bone mass through pineal-derived melatonin-MT2 pathway. J Pineal Res. CrossRefGoogle Scholar
  44. Siddiqui JA, Swarnkar G, Sharan K, Chakravarti B, Sharma G et al (2010) 8,8″-Biapigeninyl stimulates osteoblast functions and inhibits osteoclast and adipocyte functions: osteoprotective action of 8,8″-biapigeninyl in ovariectomized mice. Mol Cell Endocrinol 323:256–267. CrossRefPubMedGoogle Scholar
  45. Silver MK, Shao J, Zhu B, Chen M, Xia Y et al (2017) Prenatal naled and chlorpyrifos exposure is associated with deficits in infant motor function in a cohort of Chinese infants. Environ Int 106:248–256. CrossRefPubMedPubMedCentralGoogle Scholar
  46. Sleeman I, Che ZC, Counsell C (2016) Risk of fracture amongst patients with Parkinson’s disease and other forms of parkinsonism. Parkinsonism Relat Disord 29:60–65. CrossRefPubMedGoogle Scholar
  47. Swarnkar G, Sharan K, Siddiqui JA, Chakravarti B, Rawat P et al (2011) A novel flavonoid isolated from the steam-bark of Ulmus wallichiana planchon stimulates osteoblast function and inhibits osteoclast and adipocyte differentiation. Eur J Pharmacol 658:65–73. CrossRefPubMedGoogle Scholar
  48. Titova N, Martinez-Martin P, Katunina E, Chaudhuri KR (2017a) Advanced Parkinson’s or “complex phase” Parkinson’s disease? Re-evaluation is needed. J Neural Transm (Vienna) 124:1529–1537. CrossRefGoogle Scholar
  49. Titova N, Padmakumar C, Lewis SJG, Chaudhuri KR (2017b) Parkinson’s: a syndrome rather than a disease? J Neural Transm (Vienna) 124:907–914. CrossRefGoogle Scholar
  50. Trasande L (2017) When enough data are not enough to enact policy: the failure to ban chlorpyrifos. PLoS Biol 15:e2003671. CrossRefPubMedPubMedCentralGoogle Scholar
  51. Venerosi A, Tait S, Stecca L, Chiarotti F, De Felice A et al (2015) Effects of maternal chlorpyrifos diet on social investigation and brain neuroendocrine markers in the offspring—a mouse study. Environ Health 14:32. CrossRefPubMedPubMedCentralGoogle Scholar
  52. Watts NB (2014) Insights from the global longitudinal study of osteoporosis in women (GLOW). Nat Rev Endocrinol 10:412–422. CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Molecular NutritionCSIR-Central Food Technological Research InstituteMysuruIndia
  2. 2.Academy of Scientific and Innovative Research (AcSIR)GhaziabadIndia

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