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Cocaine-induced locomotor stimulation involves autophagic degradation of the dopamine transporter

Molecular Psychiatry volume 26, pages 370–382 (2021)Cite this article

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Abstract

Cocaine exerts its stimulant effect by inhibiting dopamine reuptake leading to increased dopamine signaling. This action is thought to reflect binding of cocaine to the dopamine transporter (DAT) to inhibit its function. However, cocaine is a relatively weak inhibitor of DAT, and many DAT inhibitors do not share the behavioral actions of cocaine. We previously showed that toxic levels of cocaine induce autophagic neuronal cell death. Here, we show that subnanomolar concentrations of cocaine elicit neural autophagy in vitro and in vivo. Autophagy inhibitors reduce the locomotor stimulant effect of cocaine in mice. Cocaine-induced autophagy degrades transporters for dopamine but not serotonin in the nucleus accumbens. Autophagy inhibition impairs cocaine conditioned place preference in mice. Our findings indicate that autophagic degradation of DAT modulates behavioral actions of cocaine.

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Fig. 1: Cocaine potently induces autophagy in neurons.
Fig. 2: Autophagy regulates the locomotor stimulant effect of cocaine.
Fig. 3: Cocaine induces autophagic degradation of the dopamine transporter (DAT).
Fig. 4: The autophagy inhibitor hydroxychloroquine impairs cocaine conditioned place preference.

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All data generated or analyzed during this study are included in this published article (and its Supplementary Data files).

References

  1. Hall FS, Sora I, Drgonova J, Li X-F, Goeb M, Uhl GR. Molecular mechanisms underlying the rewarding effects of cocaine. Ann N Y Acad Sci. 2004;1025:47–56.

    Article  CAS  PubMed  Google Scholar 

  2. Rothman RB, Baumann MH. Monoamine transporters and psychostimulant drugs. Eur J Pharm. 2003;479:23–40.

    Article  CAS  Google Scholar 

  3. Nestler EJ, Malenka RC. The addicted brain. Sci Am. 2004;290:78–85.

    Article  CAS  PubMed  Google Scholar 

  4. Edwards DJ, Bowles SK. Protein binding of cocaine in human serum. Pharm Res. 1988;5:440–2.

    Article  CAS  PubMed  Google Scholar 

  5. Guha P, Harraz MM, Snyder SH. Cocaine elicits autophagic cytotoxicity via a nitric oxide-GAPDH signaling cascade. Proc Natl Acad Sci USA. 2016;113:1417–22.

    Article  CAS  PubMed  Google Scholar 

  6. Majewska MD. Neurotoxicity and neuropathology associated with chronic cocaine abuse. NIDA Res Monogr. 1996;162:70–72.

    CAS  PubMed  Google Scholar 

  7. CLARK SL. Cellular differentiation in the kidneys of newborn mice studies with the electron microscope. J Biophys Biochem Cytol. 1957;3:349–62.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Deter RL, de Duve C. Influence of glucagon, an inducer of cellular autophagy, on some physical properties of rat liver lysosomes. J Cell Biol. 1967;33:437–49.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Dent P, Booth L, Poklepovic A, Hancock JF. Signaling alterations caused by drugs and autophagy. Cell Signal. 2019;64:109416.

    Article  CAS  PubMed  Google Scholar 

  10. Klionsky DJ, Abdelmohsen K, Abe A, Abedin MJ, Abeliovich H, Acevedo Arozena A, et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 2016;12:1–222.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Mizushima N, Yamamoto A, Matsui M, Yoshimori T, Ohsumi Y. In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker. Mol Biol Cell. 2004;15:1101–11.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Rayport S, Sulzer D, Shi WX, Sawasdikosol S, Monaco J, Batson D, et al. Identified postnatal mesolimbic dopamine neurons in culture: morphology and electrophysiology. J Neurosci. 1992;12:4264–80.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Harraz MM, Eacker SM, Wang X, Dawson TM, Dawson VL. MicroRNA-223 is neuroprotective by targeting glutamate receptors. Proc Natl Acad Sci USA. 2012;109:18962–7.

    Article  CAS  PubMed  Google Scholar 

  14. Graham JM. Isolation of lysosomes from tissues and cells by differential and density gradient centrifugation. Curr Protoc Cell Biol. 2001;Chapter 3:Unit 3.6.

  15. Tanda G, Newman AH, Ebbs AL, Tronci V, Green JL, Tallarida RJ, et al. Combinations of cocaine with other dopamine uptake inhibitors: assessment of additivity. J Pharm Exp Ther. 2009;330:802–9.

    Article  CAS  Google Scholar 

  16. Mereu M, Tronci V, Chun LE, Thomas AM, Green JL, Katz JL, et al. Cocaine-induced endocannabinoid release modulates behavioral and neurochemical sensitization in mice. Addict Biol. 2015;20:91–103.

    Article  CAS  PubMed  Google Scholar 

  17. Keighron JD, Quarterman JC, Cao J, DeMarco EM, Coggiano MA, Gleaves A, et al. Effects of (R)-modafinil and modafinil analogues on dopamine dynamics assessed by voltammetry and microdialysis in the mouse nucleus accumbens shell. ACS Chem Neurosci. 2019;10:2012–21.

    Article  CAS  PubMed  Google Scholar 

  18. Paxinos G, Franklin KBJ. The mouse brain in stereotaxic coordinates. Compact third eddition. San Diego, CA: Academic Press; 2008.

  19. Li N, Lee B, Liu R-J, Banasr M, Dwyer JM, Iwata M, et al. mTOR-dependent synapse formation underlies the rapid antidepressant effects of NMDA antagonists. Science. 2010;329:959–64.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Janowsky A, Neve K, Eshleman AJ. Uptake and release of neurotransmitters. Curr Protoc Neurosci 2001;Chapter 7:Unit7.9–7.9.22.

  21. Egan DF, Chun MGH, Vamos M, Zou H, Rong J, Miller CJ, et al. Small molecule inhibition of the autophagy kinase ULK1 and identification of ULK1 substrates. Mol Cell. 2015;59:285–97.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Lu Y, Dong S, Hao B, Li C, Zhu K, Guo W, et al. Vacuolin-1 potently and reversibly inhibits autophagosome-lysosome fusion by activating RAB5A. Autophagy. 2014;10:1895–905.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Degtyarev M, De Mazière A, Orr C, Lin J, Lee BB, Tien JY, et al. Akt inhibition promotes autophagy and sensitizes PTEN-null tumors to lysosomotropic agents. J Cell Biol. 2008;183:101–16.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Uhl GR, Hall FS, Sora I. Cocaine, reward, movement and monoamine transporters. Mol Psychiatry. 2002;7:21–26.

    Article  CAS  PubMed  Google Scholar 

  25. Maday S, Wallace KE, Holzbaur ELF. Autophagosomes initiate distally and mature during transport toward the cell soma in primary neurons. J Cell Biol. 2012;196:407–17.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Maday S, Holzbaur ELF. Autophagosome biogenesis in primary neurons follows an ordered and spatially regulated pathway. Dev Cell. 2014;30:71–85.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Hill SE, Kauffman KJ, Krout M, Richmond JE, Melia TJ, Colón-Ramos DA. Maturation and clearance of autophagosomes in neurons depends on a specific cysteine protease isoform, ATG-4.2. Dev Cell. 2019;49:251–266.e8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. WHITBY LG, HERTTING G, AXELROD J. Effect of cocaine on the disposition of noradrenaline labelled with tritium. Nature. 1960;187:604–5.

    Article  CAS  PubMed  Google Scholar 

  29. Kilty JE, Lorang D, Amara SG. Cloning and expression of a cocaine-sensitive rat dopamine transporter. Science. 1991;254:578–9.

    Article  CAS  PubMed  Google Scholar 

  30. Shimada S, Kitayama S, Lin CL, Patel A, Nanthakumar E, Gregor P, et al. Cloning and expression of a cocaine-sensitive dopamine transporter complementary DNA. Science. 1991;254:576–8.

    Article  CAS  PubMed  Google Scholar 

  31. Amara SG, Sonders MS. Neurotransmitter transporters as molecular targets for addictive drugs. Drug Alcohol Depend. 1998;51:87–96.

    Article  CAS  PubMed  Google Scholar 

  32. Ritz MC, Lamb RJ, Goldberg SR, Kuhar MJ. Cocaine receptors on dopamine transporters are related to self-administration of cocaine. Science. 1987;237:1219–23.

    Article  CAS  PubMed  Google Scholar 

  33. Ramamoorthy S, Bauman AL, Moore KR, Han H, Yang-Feng T, Chang AS, et al. Antidepressant- and cocaine-sensitive human serotonin transporter: molecular cloning, expression, and chromosomal localization. Proc Natl Acad Sci USA. 1993;90:2542–6.

    Article  CAS  PubMed  Google Scholar 

  34. Blakely RD, Bauman AL. Biogenic amine transporters: regulation in flux. Curr Opin Neurobiol. 2000;10:328–36.

    Article  CAS  PubMed  Google Scholar 

  35. Cole JO, Levin A, Beake B, Kaiser PE, Scheinbaum ML. Sibutramine: a new weight loss agent without evidence of the abuse potential associated with amphetamines. J Clin Psychopharmacol. 1998;18:231–6.

    Article  CAS  PubMed  Google Scholar 

  36. Schuh LM, Schuster CR, Hopper JA, Mendel CM. Abuse liability assessment of sibutramine, a novel weight control agent. Psychopharmacology. 2000;147:339–46.

    Article  CAS  PubMed  Google Scholar 

  37. Peck AW, Bye CE, Clubley M, Henson T, Riddington C. A comparison of bupropion hydrochloride with dexamphetamine and amitriptyline in healthy subjects. Br J Clin Pharm. 1979;7:469–78.

    Article  CAS  Google Scholar 

  38. Chait LD, Uhlenhuth EH, Johanson CE. Reinforcing and subjective effects of several anorectics in normal human volunteers. J Pharm Exp Ther. 1987;242:777–83.

    CAS  Google Scholar 

  39. Chait LD, Uhlenhuth EH, Johanson CE. The discriminative stimulus and subjective effects of phenylpropanolamine, mazindol and d-amphetamine in humans. Pharm Biochem Behav. 1986;24:1665–72.

    Article  CAS  Google Scholar 

  40. Schoedel KA, Meier D, Chakraborty B, Manniche PM, Sellers EM. Subjective and objective effects of the novel triple reuptake inhibitor tesofensine in recreational stimulant users. Clin Pharm Ther. 2010;88:69–78.

    Article  CAS  Google Scholar 

  41. Post RM, Kotin J, Goodwin FK. The effects of cocaine on depressed patients. Am J Psychiatry. 1974;131:511–7.

    Article  CAS  PubMed  Google Scholar 

  42. Uhl GR. Dopamine transporter: basic science and human variation of a key molecule for dopaminergic function, locomotion, and parkinsonism. Mov Disord. 2003;18:S71–80.

    Article  PubMed  Google Scholar 

  43. Giros B, Jaber M, Jones SR, Wightman RM, Caron MG. Hyperlocomotion and indifference to cocaine and amphetamine in mice lacking the dopamine transporter. Nature. 1996;379:606–12.

    Article  CAS  PubMed  Google Scholar 

  44. Sora I, Wichems C, Takahashi N, Li XF, Zeng Z, Revay R, et al. Cocaine reward models: conditioned place preference can be established in dopamine- and in serotonin-transporter knockout mice. Proc Natl Acad Sci USA. 1998;95:7699–704.

    Article  CAS  PubMed  Google Scholar 

  45. Sora I, Hall FS, Andrews AM, Itokawa M, Li XF, Wei HB, et al. Molecular mechanisms of cocaine reward: combined dopamine and serotonin transporter knockouts eliminate cocaine place preference. Proc Natl Acad Sci USA. 2001;98:5300–5.

    Article  CAS  PubMed  Google Scholar 

  46. Rao A, Sorkin A, Zahniser NR. Mice expressing markedly reduced striatal dopamine transporters exhibit increased locomotor activity, dopamine uptake turnover rate, and cocaine responsiveness. Synapse. 2013;67:668–77.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Tilley MR, Cagniard B, Zhuang X, Han DD, Tiao N, Gu HH. Cocaine reward and locomotion stimulation in mice with reduced dopamine transporter expression. BMC Neurosci. 2007;8:42–7.

    Article  PubMed  PubMed Central  Google Scholar 

  48. Chen R, Tilley MR, Wei H, Zhou F, Zhou F-M, Ching S, et al. Abolished cocaine reward in mice with a cocaine-insensitive dopamine transporter. Proc Natl Acad Sci USA. 2006;103:9333–8.

    Article  CAS  PubMed  Google Scholar 

  49. Hara T, Nakamura K, Matsui M, Yamamoto A, Nakahara Y, Suzuki-Migishima R, et al. Suppression of basal autophagy in neural cells causes neurodegenerative disease in mice. Nature. 2006;441:885–9.

    Article  CAS  PubMed  Google Scholar 

  50. Komatsu M, Waguri S, Chiba T, Murata S, Iwata J-I, Tanida I, et al. Loss of autophagy in the central nervous system causes neurodegeneration in mice. Nature. 2006;441:880–4.

    Article  CAS  PubMed  Google Scholar 

  51. Valente MJ, Amaral C, Correia-da-Silva G, Duarte JA, Bastos M, de L, et al. Methylone and MDPV activate autophagy in human dopaminergic SH-SY5Y cells: a new insight into the context of β-keto amphetamines-related neurotoxicity. Arch Toxicol. 2017;91:3663–76.

    Article  CAS  PubMed  Google Scholar 

  52. Mercer LD, Higgins GC, Lau CL, Lawrence AJ, Beart PM. MDMA-induced neurotoxicity of serotonin neurons involves autophagy and rilmenidine is protective against its pathobiology. Neurochem Int. 2017;105:80–90.

    Article  PubMed  Google Scholar 

  53. Larsen KE, Fon EA, Hastings TG, Edwards RH, Sulzer D. Methamphetamine-induced degeneration of dopaminergic neurons involves autophagy and upregulation of dopamine synthesis. J Neurosci. 2002;22:8951–60.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank L. Hester, R. Barrow, A. Snowman, S. McTeer, and L. Albacarys from the SHS laboratory for their assistance. We thank B. Smith and the Microscope Core Facility at the Institute for Basic Biomedical Sciences, Johns Hopkins School of Medicine, for helping in preparation of the TEM samples. We are also grateful for fruitful discussions with members of the SHS Laboratory.

Funding

This work was supported by U.S. Public Health Service Grants DA00266 and DA044123 to SHS and a NARSAD Young Investigator Grant (# 25360) from the Brain & Behavior Foundation to MMH. Support for this research was provided in part by the National Institute on Drug Abuse—Intramural Research Program, NIH/DHHS (Z1A DA000611).

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Author notes

  1. These authors contributed equally: Maged M. Harraz, Prasun Guha, Evan R. Semenza, Adarsha P. Malla

Authors and Affiliations

  1. The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

    Maged M. Harraz, Prasun Guha, In Guk Kang, Evan R. Semenza, Adarsha P. Malla, Young Jun Song, Luke Reilly, Isaac Treisman, Pedro Cortés & Solomon H. Snyder

  2. Medication Development Program, Molecular Targets and Medications Discovery Branch, National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, MD, 21224, USA

    Mark A. Coggiano & Gianluigi Tanda

  3. Department of Neurology and Johns Hopkins Drug Discovery (JHDD) Program, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

    Vijayabhaskar Veeravalli & Rana Rais

  4. Department of Psychiatry and Behavioral Sciences, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

    Solomon H. Snyder

  5. Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

    Solomon H. Snyder

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Contributions

MMH, PG, PC, IGK, ERS, YJS, LR, IT, APM, MAC, and VV performed experiments. MMH, PG, GT, RR, and SHS designed experiments, analyzed data, and wrote the manuscript.

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Correspondence to Solomon H. Snyder.

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Harraz, M.M., Guha, P., Kang, I.G. et al. Cocaine-induced locomotor stimulation involves autophagic degradation of the dopamine transporter. Mol Psychiatry 26, 370–382 (2021). https://doi.org/10.1038/s41380-020-00978-y

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