Spatial memory deficits in mice induced by chemotherapeutic agents are prevented by acetylcholinesterase inhibitors

  • Rex M. PhilpotEmail author
  • M. Ficken
  • B. E. Johns
  • M. E. Engberg
  • L. Wecker
Original Article



These studies determined whether the acetylcholinesterase inhibitors, donepezil and galantamine, both of which are approved for the treatment of cognitive deficits in Alzheimer’s disease, can prevent or reverse spatial memory deficits in mice induced by cyclophosphamide and doxorubicin, cytotoxic agents commonly used to treat breast cancer.


Female BALB/C mice were trained in the Morris water maze to identify the location of a submerged platform, and, following baseline assessment of spatial memory, received injections of cyclophosphamide and doxorubicin once per week for 4 weeks to impair spatial memory. Saline or acetylcholinesterase inhibitors were administered daily either concurrent with the chemotherapy injections (prevention) or beginning 1 week following the final chemotherapy injections (reversal), and spatial memory was assessed weekly.


Spatial memory declined during and following weekly injections of cyclophosphamide and doxorubicin, and was unaltered when the acetylcholinesterase inhibitors were administered following the manifestation of chemotherapy-induced deficits. In contrast, spatial memory of mice receiving the acetylcholinesterase inhibitors concurrent with chemotherapy did not differ from that at baseline.


Results indicate that chemotherapy-induced spatial memory deficits in mice can be prevented, but not reversed by the use of acetylcholinesterase inhibitors concomitant with chemotherapy, suggesting that these agents should be investigated further for the prevention of chemobrain.


Chemobrain Cyclophosphamide Donepezil Doxorubicin Galantamine 



The authors would like to thank April Lindon for her technical assistance during this study and Dr. Heather Jim for her valuable suggestions and feedback.


These studies were supported in part by USF Research and Innovation Proposal Enhancement Grant and the American Cancer Society Institutional Research Grant, Moffitt Cancer Center.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

The care and use of animals were approved in accordance with guidelines set by the University of South Florida Institutional Animal Care and Use Committee, the National Institutes of Health Guide for the Care and Use of Laboratory Animals and the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines of the National Centre for the Replacement, Refinement and Reduction of Animals in Research.


  1. 1.
    Wieneke MH, Dienst ER (1995) Neuropsychological assessment of cognitive functioning following chemotherapy for breast cancer. Psychooncology 4:61–66CrossRefGoogle Scholar
  2. 2.
    Wefel JS, Lenzi R, Theriault RL, Davis RN, Meyers CA (2004) The cognitive sequelae of standard-dose adjuvant chemotherapy in women with breast carcinoma: results of a prospective, randomized, longitudinal trial. Cancer 100(11):2292–2299. CrossRefGoogle Scholar
  3. 3.
    Ahles TA, Root JC, Ryan EL (2012) Cancer- and cancer treatment-associated cognitive change: an update on the state of the science. J Clin Oncol 30(30):3675–3686. CrossRefGoogle Scholar
  4. 4.
    Tannock IF, Ahles TA, Ganz PA, Van Dam FS (2004) Cognitive impairment associated with chemotherapy for cancer: report of a workshop. J Clin Oncol 22(11):2233–2239. CrossRefGoogle Scholar
  5. 5.
    Kohli S, Griggs JJ, Roscoe JA, Jean-Pierre P, Bole C, Mustian KM, Hill R, Smith K, Gross H, Morrow GR (2007) Self-reported cognitive impairment in patients with cancer. J Oncol Pract 3(2):54–59. CrossRefGoogle Scholar
  6. 6.
    Wefel JS, Saleeba AK, Buzdar AU, Meyers CA (2010) Acute and late onset cognitive dysfunction associated with chemotherapy in women with breast cancer. Cancer 116(14):3348–3356. CrossRefGoogle Scholar
  7. 7.
    Raffa RB (2010) Is a picture worth a thousand (forgotten) words?: neuroimaging evidence for the cognitive deficits in ‘chemo-fog’/’chemo-brain’. J Clin Pharm Ther 35(1):1–9. CrossRefGoogle Scholar
  8. 8.
    Koppelmans V, Breteler MM, Boogerd W, Seynaeve C, Gundy C, Schagen SB (2012) Neuropsychological performance in survivors of breast cancer more than 20 years after adjuvant chemotherapy. J Clin Oncol 30(10):1080–1086. CrossRefGoogle Scholar
  9. 9.
    Wilkinson DG, Francis PT, Schwam E, Payne-Parrish J (2004) Cholinesterase inhibitors used in the treatment of Alzheimer’s disease: the relationship between pharmacological effects and clinical efficacy. Drugs Aging 21(7):453–478CrossRefGoogle Scholar
  10. 10.
    Ribeiz SR, Bassitt DP, Arrais JA, Avila R, Steffens DC, Bottino CM (2010) Cholinesterase inhibitors as adjunctive therapy in patients with schizophrenia and schizoaffective disorder: a review and meta-analysis of the literature. CNS Drugs 24(4):303–317. CrossRefGoogle Scholar
  11. 11.
    Everitt BJ, Robbins TW (1997) Central cholinergic systems and cognition. Annu Rev Psychol 48:649–684. CrossRefGoogle Scholar
  12. 12.
    Lawrence JA, Griffin L, Balcueva EP, Groteluschen DL, Samuel TA, Lesser GJ, Naughton MJ, Case LD, Shaw EG, Rapp SR (2016) A study of donepezil in female breast cancer survivors with self-reported cognitive dysfunction 1 to 5 years following adjuvant chemotherapy. J Cancer Surviv 10(1):176–184. CrossRefGoogle Scholar
  13. 13.
    Winocur G, Binns MA, Tannock I (2011) Donepezil reduces cognitive impairment associated with anti-cancer drugs in a mouse model. Neuropharmacology 61(8):1222–1228. CrossRefGoogle Scholar
  14. 14.
    Macleod JE, DeLeo JA, Hickey WF, Ahles TA, Saykin AJ, Bucci DJ (2007) Cancer chemotherapy impairs contextual but not cue-specific fear memory. Behav Brain Res 181(1):168–172. CrossRefGoogle Scholar
  15. 15.
    Konat GW, Kraszpulski M, James I, Zhang HT, Abraham J (2008) Cognitive dysfunction induced by chronic administration of common cancer chemotherapeutics in rats. Metab Brain Dis 23(3):325–333. CrossRefGoogle Scholar
  16. 16.
    Philpot RM, Ficken M, Wecker L (2016) Doxorubicin and cyclophosphamide lead to long-lasting impairment of spatial memory in female, but not male mice. Behav Brain Res 307:165–175. CrossRefGoogle Scholar
  17. 17.
    Rendeiro C, Sheriff A, Bhattacharya TK, Gogola JV, Baxter JH, Chen H, Helferich WG, Roy EJ, Rhodes JS (2016) Long-lasting impairments in adult neurogenesis, spatial learning and memory from a standard chemotherapy regimen used to treat breast cancer. Behav Brain Res 315:10–22. CrossRefGoogle Scholar
  18. 18.
    Samochocki M, Hoffle A, Fehrenbacher A, Jostock R, Ludwig J, Christner C, Radina M, Zerlin M, Ullmer C, Pereira EF, Lubbert H, Albuquerque EX, Maelicke A (2003) Galantamine is an allosterically potentiating ligand of neuronal nicotinic but not of muscarinic acetylcholine receptors. J Pharmacol Exp Ther 305(3):1024–1036. CrossRefGoogle Scholar
  19. 19.
    Thomsen MS, Hansen HH, Timmerman DB, Mikkelsen JD (2010) Cognitive improvement by activation of alpha7 nicotinic acetylcholine receptors: from animal models to human pathophysiology. Curr Pharm Des 16(3):323–343CrossRefGoogle Scholar
  20. 20.
    Yang M, Kim JS, Song MS, Kim SH, Kang SS, Bae CS, Kim JC, Wang H, Shin T, Moon C (2010) Cyclophosphamide impairs hippocampus-dependent learning and memory in adult mice: possible involvement of hippocampal neurogenesis in chemotherapy-induced memory deficits. Neurobiol Learn Mem 93(4):487–494. CrossRefGoogle Scholar
  21. 21.
    Christie LA, Acharya MM, Parihar VK, Nguyen A, Martirosian V, Limoli CL (2012) Impaired cognitive function and hippocampal neurogenesis following cancer chemotherapy. Clin Cancer Res 18(7):1954–1965. CrossRefGoogle Scholar
  22. 22.
    Kitamura Y, Hattori S, Yoneda S, Watanabe S, Kanemoto E, Sugimoto M, Kawai T, Machida A, Kanzaki H, Miyazaki I, Asanuma M, Sendo T (2015) Doxorubicin and cyclophosphamide treatment produces anxiety-like behavior and spatial cognition impairment in rats: possible involvement of hippocampal neurogenesis via brain-derived neurotrophic factor and cyclin D1 regulation. Behav Brain Res 292:184–193. CrossRefGoogle Scholar
  23. 23.
    Mustafa S, Walker A, Bennett G, Wigmore PM (2008) 5-Fluorouracil chemotherapy affects spatial working memory and newborn neurons in the adult rat hippocampus. Eur J Neurosci 28(2):323–330. CrossRefGoogle Scholar
  24. 24.
    Lyons L, ElBeltagy M, Umka J, Markwick R, Startin C, Bennett G, Wigmore P (2011) Fluoxetine reverses the memory impairment and reduction in proliferation and survival of hippocampal cells caused by methotrexate chemotherapy. Psychopharmacology (Berl) 215(1):105–115. CrossRefGoogle Scholar
  25. 25.
    Morris RG, Garrud P, Rawlins JN, O’Keefe J (1982) Place navigation impaired in rats with hippocampal lesions. Nature 297(5868):681–683CrossRefGoogle Scholar
  26. 26.
    Beiko J, Lander R, Hampson E, Boon F, Cain DP (2004) Contribution of sex differences in the acute stress response to sex differences in water maze performance in the rat. Behav Brain Res 151(1–2):239–253. CrossRefGoogle Scholar
  27. 27.
    Huang Y, Zhou W, Zhang Y (2012) Bright lighting conditions during testing increase thigmotaxis and impair water maze performance in BALB/c mice. Behav Brain Res 226(1):26–31. CrossRefGoogle Scholar
  28. 28.
    Kilkenny C, Browne WJ, Cuthill IC, Emerson M, Altman DG (2010) Improving bioscience research reporting: the ARRIVE guidelines for reporting animal research. J Pharmacol Pharmacother 1(2):94–99. CrossRefGoogle Scholar
  29. 29.
    Jenkins V, Shilling V, Deutsch G, Bloomfield D, Morris R, Allan S, Bishop H, Hodson N, Mitra S, Sadler G, Shah E, Stein R, Whitehead S, Winstanley J (2006) A 3-year prospective study of the effects of adjuvant treatments on cognition in women with early stage breast cancer. Br J Cancer 94(6):828–834. CrossRefGoogle Scholar
  30. 30.
    Wefel JS, Witgert ME, Meyers CA (2008) Neuropsychological sequelae of non-central nervous system cancer and cancer therapy. Neuropsychol Rev 18(2):121–131. CrossRefGoogle Scholar
  31. 31.
    Francis DD, Zaharia MD, Shanks N, Anisman H (1995) Stress-induced disturbances in Morris water-maze performance: interstrain variability. Physiol Behav 58(1):57–65CrossRefGoogle Scholar
  32. 32.
    Wahlsten D, Cooper SF, Crabbe JC (2005) Different rankings of inbred mouse strains on the Morris maze and a refined 4-arm water escape task. Behav Brain Res 165(1):36–51. CrossRefGoogle Scholar
  33. 33.
    Castagne V, Moser P, Roux S, Porsolt RD (2011) Rodent models of depression: forced swim and tail suspension behavioral despair tests in rats and mice. Curr Protoc Neurosci. (Chapter 8:Unit 8 10A) Google Scholar
  34. 34.
    Gergalova G, Lykhmus O, Komisarenko S, Skok M (2014) α7 nicotinic acetylcholine receptors control cytochrome c release from isolated mitochondria through kinase-mediated pathways. Int J Biochem Cell Biol 49:26–31. CrossRefGoogle Scholar
  35. 35.
    Genka S, Deutsch J, Stahle PL, Shetty UH, John V, Robinson C, Rapoport SI, Greig NH (1990) Brain and plasma pharmacokinetics and anticancer activities of cyclophosphamide and phosphoramide mustard in the rat. Cancer Chemother Pharmacol 27(1):1–7CrossRefGoogle Scholar
  36. 36.
    von Holst H, Knochenhauer E, Blomgren H, Collins VP, Ehn L, Lindquist M, Noren G, Peterson C (1990) Uptake of adriamycin in tumour and surrounding brain tissue in patients with malignant gliomas. Acta Neurochir (Wien) 104(1–2):13–16CrossRefGoogle Scholar
  37. 37.
    Janelsins MC, Mustian KM, Palesh OG, Mohile SG, Peppone LJ, Sprod LK, Heckler CE, Roscoe JA, Katz AW, Williams JP, Morrow GR (2012) Differential expression of cytokines in breast cancer patients receiving different chemotherapies: implications for cognitive impairment research. Support Care Cancer 20(4):831–839. CrossRefGoogle Scholar
  38. 38.
    Cheung YT, Ng T, Shwe M, Ho HK, Foo KM, Cham MT, Lee JA, Fan G, Tan YP, Yong WS, Madhukumar P, Loo SK, Ang SF, Wong M, Chay WY, Ooi WS, Dent RA, Yap YS, Ng R, Chan A (2015) Association of proinflammatory cytokines and chemotherapy-associated cognitive impairment in breast cancer patients: a multi-centered, prospective, cohort study. Ann Oncol 26(7):1446–1451. CrossRefGoogle Scholar
  39. 39.
    Wilson CJ, Finch CE, Cohen HJ (2002) Cytokines and cognition–the case for a head-to-toe inflammatory paradigm. J Am Geriatr Soc 50(12):2041–2056CrossRefGoogle Scholar
  40. 40.
    Kelley KW, Bluthe RM, Dantzer R, Zhou JH, Shen WH, Johnson RW, Broussard SR (2003) Cytokine-induced sickness behavior. Brain Behav Immun 17(Suppl 1):S112–S118CrossRefGoogle Scholar
  41. 41.
    Kawashima K, Fujii T (2008) Basic and clinical aspects of non-neuronal acetylcholine: overview of non-neuronal cholinergic systems and their biological significance. J Pharmacol Sci 106(2):167–173CrossRefGoogle Scholar
  42. 42.
    Tracey KJ (2007) Physiology and immunology of the cholinergic antiinflammatory pathway. J Clin Investig 117(2):289–296. CrossRefGoogle Scholar
  43. 43.
    Wang DW, Zhou RB, Yao YM (2009) Role of cholinergic anti-inflammatory pathway in regulating host response and its interventional strategy for inflammatory diseases. Chin J Traumatol 12(6):355–364Google Scholar
  44. 44.
    Carnevale D, De Simone R, Minghetti L (2007) Microglia-neuron interaction in inflammatory and degenerative diseases: role of cholinergic and noradrenergic systems. CNS Neurol Disord Drug Targ 6(6):388–397CrossRefGoogle Scholar
  45. 45.
    Tsvetkova D, Obreshkova D, Zheleva-Dimitrova D, Saso L (2013) Antioxidant activity of galantamine and some of its derivatives. Curr Med Chem 20(36):4595–4608CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Psychiatry and Behavioral Neurosciences, Morsani College of MedicineUniversity of South FloridaTampaUSA

Personalised recommendations