Abstract
Neuronal cyclin-dependent kinase-5 (Cdk5) and its neuron-specific activator p35 play a major role in regulating the cytoskeleton dynamics. Since opioid addiction was associated with hyperphosphorylation of neurofilament (NF) in postmortem human brains, this study was undertaken to assess the status of the cdk5/p35 complex and its relation with NF-H phosphorylation in brains of chronic opioid abusers. Decreased immunodensities of cdk5 (18%) and p35 (26–44%) were found in the prefrontal cortex of opioid addicts compared with matched controls. In the same brains, the densities of p25 (a truncated neurotoxic form of p35), phosphatase PP2Ac and μ-calpain were found unaltered. Acute treatment of rats with morphine (30 mg/kg, 2 h) increased the density of cdk5 (35%), but not that of p35, in the cerebral cortex. In contrast, chronic morphine (10–100 mg/kg for 5 days) induced marked decreases in cdk5 (40%) and p35 (47%) in rat brain. In brains of opioid addicts, the density of phosphorylated NF-H was increased (43%) as well as the ratio of phosphorylated to nonphosphorylated NF-H forms (two-fold). In these brains, phosphorylated NF-H significantly correlated with p35 (r=0.58) but not with cdk5 (r=0.03). The results suggest that opiate addiction is associated with downregulation of cdk5/p35 levels in the brain. This downregulation and the aberrant hyperphosphorylation of NF-H proteins might have important consequences in the development of neural plasticity associated with opiate addiction in humans.
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References
Bibb JA, Chen J, Taylor JR, Svenningsson P, Nishi A, Snyder GL (2001). Effects of chronic exposure to cocaine are regulated by the neuronal protein Cdk5. Nature 410: 376–380.
Dhavan R, Tsai L-H (2001). A decade of CDK5. Nature Rev/Mol Cell Biol 2: 749–759.
Ferrer-Alcón F, García-Sevilla JA, La Harpe R, Walzer C, Guimón J (1999). Regulation of p35/cyclin-dependent kinase 5 (p35/cdk-5) complex in brains of opioid addicts. S07-10, p 49.
Ferrer-Alcón F, García-Sevilla JA, Jaquet Ph, La Harpe R, Riederer BM, Walzer C et al (2000). Regulation of nonphosphorylated and phosphorylated forms of neurofilament proteins in the prefrontal cortex of human opioid addicts. J Neurosci Res 61: 338–349.
Fountoulakis M, Hardmeier R, Höger H, Lubec G (2001). Postmortem changes in the level of brain proteins. Exp Neurol 167: 86–94.
García-Sevilla JA, Ventayol P, Busquets X, La Harpe R, Walzer C, Guimón J (1997). Marked decrease of immunolabelled 68 kDa neurofilament (NF-L) proteins in brains of opiate addicts. NeuroReport 8: 1561–1570.
Goldberger BA, Cone EJ, Grant T, Caplan YH, Levine BS, Smialek JE (1994). Disposition of heroin and its metabolites in heroin-related deaths. J Anal Toxicol 18: 22–28.
Grant P, Sharma P, Pant HC (2001). Cyclin-dependent protein kinase 5 (CDK5) and the regulation of neurofilament metabolism. Eur J Biochem 268: 1534–1546.
Hellmich MR, Pant HC, Wada E, Battey JF (1992). Neuronal cdc2-like kinase: a CDC2-related protein kinase with predominantly neuronal expression. Proc Natl Acad Sci USA 89: 10867–10871.
Jaquet PhE, Ferrer-Alcón M, Ventayol P, Guimón J, García-Sevilla JA (2001). Acute and chronic effects of morphine and naloxone on the phosphorylation of neurofilament-H proteins in the rat brain. Neurosci Lett 304: 37–40.
Kusakawa G-I, Saito T, Onuki R, Ishiguro K, Kishimoto T, Hisanaga S-I (2000). Calpain-dependent proteolytic cleavage of the p35 cyclin-dependent kinase 5 activator to p25. J Biol Chem 275: 17166–17172.
Law P-Y, Wong YH, Loh HH (2000). Molecular mechanisms and regulation of opioid receptor signaling. Annu Rev Pharmacol 40: 389–430.
Lee M-S, Kwon YT, Li M, Peng J, Friedlander RM, Tsai L-H (2000). Neurotoxicity induces cleavage of p35 to p25 by calpain. Nature 405: 360–364.
Lew J, Huang Q-Q, Qi Z, Winkfein RJ, Aebersold R, Hunt T et al (1994). A brain-specific activator of cyclin-dependent kinase 5. Nature 371: 423–426.
Maccioni RB, Otth C, Concha II, Muñoz JP (2001). The protein kinase Cdk5. Structural aspects, roles in neurogenesis and involvement in Alzheimer's pathology. Eur J Biochem 268: 1518–1527.
McLeman ER, Warsh JJ, Ang L, Li PP, Kalasinsky KS, Ross BM et al (2000). The human nucleus accumbens is highly susceptible to G protein down-regulation by methamphetamine and heroin. J Neurochem 74: 2120–2126.
Meana JJ, González-Maeso J, García-Sevilla JA, Guimón J (2000). μ -opioid receptor and α2-adrenoceptor agonist stimulation of 35SGTPγS binding to G-proteins in postmortem brains of opioid addicts. Mol Psychiatry 5: 308–315.
Merril J, Garvey T, Rosson C (1996). Methadone concentrations taken as indicating deaths due to overdose need to be reviewed. Br Med J 313: 1481.
Nakamura S, Kawamoto Y, Nakano N, Akiguchi I, Kimura J (1997). p35nck5a and cyclin-dependent kinase 5 colocalize in Lewy bodies of brains with Parkinson's disease. Acta Nueropathol 94: 153–157.
Nestler EJ (2001). Molecular basis of long-term plasticity underlying addiction. Nat Rev/Neurosci 2: 119–128.
Nestler EJ, Barrot M, Self DW (2001). ΔFosB: a sustained molecular switch for addiction. Proc Natl Acad Sci USA 98: 11042–11046.
Nye HE, Nestler EJ (1996). Induction of chronic Fos-related antigens in rat brain by chronic morphine administration. Mol Pharmacol 49: 636–645.
Pant AC, Veeranna GJ, Pant HC, Amin N (1997). Phosphorylation of human high molecular weight neurofilament protein (hNF-H) by neuronal cyclin-dependent kinase 5 (cdk5). Brain Res 765: 259–266.
Patrick GN, Zhou P, Kwon YT, Howley PM, Tsai L-H (1998). p35, the neuronal-specific activator of cyclin-dependent kinase 5 (Cdk5) is degraded by the ubiquitin-proteasome pathway. J Biol Chem 273: 24057–24064.
Patrick GN, Zukerberg L, Nikolic M, de la Monte S, Dikkes P, Tsai L-H (1999). Conversion of p35 to p25 deregulates Cdk5 activity and promotes neurodegeneration. Nature 402: 615–622.
Qi Z, Tang D, Zhu X, Fujita DJ, Wang JH (1998). Association of neurofilament proteins with neuronal Cdk5 activator. J Biol Chem 273: 2329–2335.
Sastre M, García-Sevilla JA (1994). Density of alpha-2A adrenoceptors and Gi proteins in the human brain: ratio of high affinity agonist sites to antagonist sites and effect of age. J Pharmacol Exp Ther 269: 1062–1072.
Sun D, Leung CL, Liem RKH (1996). Phosphorylation of the high molecular weight neurofilament protein (NF-H) by Cdk5 and p35. J Biol Chem 271: 14245–14251.
Tagliaro F, De Battisti Z, Smith FP, Marigo M (1998). Death from heroin overdose: findings from hair analysis. Lancet 351: 1923–1925.
Tang D, Wang JH (1996). Cyclin-dependent kinase 5 (Cdk5) and neuron-specific Cdk5 activators. Prog Cell Cycle Res 2: 205–216.
Tsai L-H, Delalle I, Caviness Jr VS, Chae T, Harlow E (1994). p35 is a neural-specific regulatory subunit of cyclin-dependent kinase 5. Nature 371: 419–423.
Veeranna GJ, Shetty KT, Link WT, Jaffe H, Wang J, Pant HC (1995). Neuronal cyclin-dependent kinase-5 phosphorylation sites in neurofilament protein (NF-H) are dephosphorylated by protein phosphatase 2A. J Neurochem 64: 2681–2690.
Veeranna GJ, Shetty KT, Takahashi M, Grant Ph, Pant HC (2000). Cdk5 and MAPK are associated with complexes of cytoskeletal proteins in rat brain. Mol Brain Res 76: 229–236.
Ventayol P, Busquets X, García-Sevilla JA (1997). Modulation of immunoreactive protein kinase C-α and β isoforms and G proteins by acute and chronic treatments with morphine and other opiate drugs in rat brain. Naunyn-Schmiedeberg's Arch Pharmacol 355: 491–500.
Zheng M, Leung CL, Liem RK (1998). Region-specific expression of cyclin-dependent kinase 5 (Cdk5) and its activators, p35 and p39, in the developing and adult rat central nervous system. J Neurobiol 35: 141–159.
Acknowledgements
This study was supported by Grant 32–57066.99 (JAG-S) from the Fonds National Suisse de la Recherche Scientifique (FNSRS, Bern, Switzerland), and by Grant BFI 2000–0306 (JAG-S) from the Ministerio de Ciencia y Tecnología (MCT, Madrid, Spain). MF-A was supported by a predoctoral fellowship from the FNSRS. JAG-S is a member of the Institut d'Estudis Catalans (Barcelona, Spain).
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Ferrer-Alcón, M., La Harpe, R., Guimón, J. et al. Downregulation of Neuronal cdk5/p35 in Opioid Addicts and Opiate-Treated Rats: Relation to Neurofilament Phosphorylation. Neuropsychopharmacol 28, 947–955 (2003). https://doi.org/10.1038/sj.npp.1300095
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DOI: https://doi.org/10.1038/sj.npp.1300095
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