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CDK5 Contributes to Neuronal Apoptosis via Promoting MEF2D Phosphorylation in Rat Model of Intracerebral Hemorrhage

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Abstract

Cyclin-dependent kinase-5 (CDK5), a serine/threonine kinase which can be activated by its neuron-specific activator p35, or its truncated form p25, plays an important role in a variety of neuronal events, including neuronal migration, synaptic transmission, and neuronal death. Accumulating evidence has shown that abnormal activation of CDK5 was a critical neuronal pro-death signal in central nervous system (CNS) diseases. However, it remains unclear how CDK5 functions upon neuronal apoptosis following intracerebral hemorrhage (ICH). In the present study, we established ICH models by injecting autologous whole blood into the right basal ganglia of adult rats and assessed their neurological deficits by behavioral tests. CDK5 protein levels and kinase activities were upregulated adjacent to the hematoma following ICH. Immunofluorescent staining showed CDK5 was mainly localized in neurons, rather than in astrocytes or oligodendrocytes. Furthermore, active caspase-3, an apoptotic marker, showed a temporally parallel expression with the protein levels/kinase activities of CDK5 following ICH. Meantime, myocyte enhancer factor 2D (MEF2D), a pro-survival transcription factor which could be phosphorylated inactivation by CDK5, also exhibited high phosphorylation levels following ICH. In vitro, we obtained a consistent upregulation of CDK5 kinase activity in primary cortical neurons after thrombin treatment. Knocking down CDK5 kinase activity suppressed neuronal apoptosis and coupled with reduced MEF2D phosphorylation at ser444 residues. Thus, we speculated that CDK5 might exert an important function in the regulation of neuronal apoptosis following ICH.

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References

  • Barros-Minones L, Martin-de-Saavedra D, Perez-Alvarez S, Orejana L, Suquia V, Goni-Allo B, Hervias I, Lopez MG, Jordan J, Aguirre N, Puerta E (2013) Inhibition of calpain-regulated p35/cdk5 plays a central role in sildenafil-induced protection against chemical hypoxia produced by malonate. Biochim Biophys Acta 1832(6):705–717

    Article  CAS  PubMed  Google Scholar 

  • Bhattacharjee RN, Banks GC, Trotter KW, Lee HL, Archer TK (2001) Histone H1 phosphorylation by Cdk2 selectively modulates mouse mammary tumor virus transcription through chromatin remodeling. Mol Cell Biol 21(16):5417–5425

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Crandall KM, Rost NS, Sheth KN (2011) Prognosis in intracerebral hemorrhage. Rev Neurol Dis 8(1–2):23–29

    PubMed  Google Scholar 

  • Dhavan R, Tsai LH (2001) A decade of CDK5. Nat Rev Mol Cell Biol 2(10):749–759

    Article  CAS  PubMed  Google Scholar 

  • Falcone GJ, Biffi A, Devan WJ, Brouwers HB, Anderson CD, Valant V, Ayres AM, Schwab K, Rost NS, Goldstein JN, Viswanathan A, Greenberg SM, Selim M, Meschia JF, Brown DL, Worrall BB, Silliman SL, Tirschwell DL, Rosand J (2013) Burden of blood pressure-related alleles is associated with larger hematoma volume and worse outcome in intracerebral hemorrhage. Stroke 44(2):321–326

    Article  PubMed Central  PubMed  Google Scholar 

  • Giannoulias D, Patel FA, Holloway AC, Lye SJ, Tai HH, Challis JR (2002) Differential changes in 15-hydroxyprostaglandin dehydrogenase and prostaglandin H synthase (types I and II) in human pregnant myometrium. J Clin Endocrinol Metab 87(3):1345–1352

    Article  CAS  PubMed  Google Scholar 

  • Gomes JA, Manno E (2013) New developments in the treatment of intracerebral hemorrhage. Neurol Clin 31(3):721–735

    Article  PubMed  Google Scholar 

  • Gong X, Tang X, Wiedmann M, Wang X, Peng J, Zheng D, Blair LA, Marshall J, Mao Z (2003) Cdk5-mediated inhibition of the protective effects of transcription factor MEF2 in neurotoxicity-induced apoptosis. Neuron 38(1):33–46

    Article  CAS  PubMed  Google Scholar 

  • Hua Y, Schallert T, Keep RF, Wu J, Hoff JT, Xi G (2002) Behavioral tests after intracerebral hemorrhage in the rat. Stroke 33(10):2478–2484

    Article  PubMed  Google Scholar 

  • Hwang BY, Appelboom G, Ayer A, Kellner CP, Kotchetkov IS, Gigante PR, Haque R, Kellner M, Connolly ES (2011) Advances in neuroprotective strategies: potential therapies for intracerebral hemorrhage. Cerebrovasc Dis 31(3):211–222

    Article  PubMed Central  PubMed  Google Scholar 

  • Karabiyikoglu M, Hua Y, Keep RF, Ennis SR, Xi G (2004) Intracerebral hirudin injection attenuates ischemic damage and neurologic deficits without altering local cerebral blood flow. J Cereb Blood Flow Metab 24(2):159–166

    Article  CAS  PubMed  Google Scholar 

  • Ke K, Li L, Rui Y, Zheng H, Tan X, Xu W, Cao J, Xu J, Cui G, Xu G, Cao M (2013) Increased expression of small heat shock protein alphaB-crystallin after intracerebral hemorrhage in adult rats. J Mol Neurosci 51(1):159–169

    Article  CAS  PubMed  Google Scholar 

  • Kesavapany S, Amin N, Zheng YL, Nijhara R, Jaffe H, Sihag R, Gutkind JS, Takahashi S, Kulkarni A, Grant P, Pant HC (2004) p35/cyclin-dependent kinase 5 phosphorylation of ras guanine nucleotide releasing factor 2 (RasGRF2) mediates Rac-dependent extracellular signal-regulated kinase 1/2 activity, altering RasGRF2 and microtubule-associated protein 1b distribution in neurons. J Neurosci 24(18):4421–4431

    Article  CAS  PubMed  Google Scholar 

  • Li M, Linseman DA, Allen MP, Meintzer MK, Wang X, Laessig T, Wierman ME, Heidenreich KA (2001) Myocyte enhancer factor 2A and 2D undergo phosphorylation and caspase-mediated degradation during apoptosis of rat cerebellar granule neurons. J Neurosci 21(17):6544–6552

    CAS  PubMed  Google Scholar 

  • Li BS, Zhang L, Takahashi S, Ma W, Jaffe H, Kulkarni AB, Pant HC (2002) Cyclin-dependent kinase 5 prevents neuronal apoptosis by negative regulation of c-Jun N-terminal kinase 3. EMBO J 21(3):324–333

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Li BS, Ma W, Jaffe H, Zheng Y, Takahashi S, Zhang L, Kulkarni AB, Pant HC (2003) Cyclin-dependent kinase-5 is involved in neuregulin-dependent activation of phosphatidylinositol 3-kinase and Akt activity mediating neuronal survival. J Biol Chem 278(37):35702–35709

    Article  CAS  PubMed  Google Scholar 

  • Lopes JP, Agostinho P (2011) Cdk5: multitasking between physiological and pathological conditions. Prog Neurobiol 94(1):49–63

    Article  CAS  PubMed  Google Scholar 

  • Ma Y, Bao J, Zhao X, Shen H, Lv J, Ma S, Zhang X, Li Z, Wang S, Wang Q, Ji J (2013) Activated cyclin-dependent kinase 5 promotes microglial phagocytosis of fibrillar beta-amyloid by up-regulating lipoprotein lipase expression. Mol Cell Proteomics 12(10):2833–2844

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Mao Z, Bonni A, Xia F, Nadal-Vicens M, Greenberg ME (1999) Neuronal activity-dependent cell survival mediated by transcription factor MEF2. Science 286(5440):785–790

    Article  CAS  PubMed  Google Scholar 

  • Nguyen MD, Boudreau M, Kriz J, Couillard-Despres S, Kaplan DR, Julien JP (2003) Cell cycle regulators in the neuronal death pathway of amyotrophic lateral sclerosis caused by mutant superoxide dismutase 1. J Neurosci 23(6):2131–2140

    CAS  PubMed  Google Scholar 

  • Nikolic M, Dudek H, Kwon YT, Ramos YF, Tsai LH (1996) The cdk5/p35 kinase is essential for neurite outgrowth during neuronal differentiation. Genes Dev 10(7):816–825

    Article  CAS  PubMed  Google Scholar 

  • O’Hare MJ, Kushwaha N, Zhang Y, Aleyasin H, Callaghan SM, Slack RS, Albert PR, Vincent I, Park DS (2005) Differential roles of nuclear and cytoplasmic cyclin-dependent kinase 5 in apoptotic and excitotoxic neuronal death. J Neurosci 25(39):8954–8966

    Article  PubMed  Google Scholar 

  • Okamoto S, Li Z, Ju C, Scholzke MN, Mathews E, Cui J, Salvesen GS, Bossy-Wetzel E, Lipton SA (2002) Dominant-interfering forms of MEF2 generated by caspase cleavage contribute to NMDA-induced neuronal apoptosis. Proc Natl Acad Sci U S A 99(6):3974–3979

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Otyepka M, Bartova I, Kriz Z, Koca J (2006) Different mechanisms of CDK5 and CDK2 activation as revealed by CDK5/p25 and CDK2/cyclin a dynamics. J Biol Chem 281(11):7271–7281

    Article  CAS  PubMed  Google Scholar 

  • Patrick GN, Zukerberg L, Nikolic M, de la Monte S, Dikkes P, Tsai LH (1999) Conversion of p35 to p25 deregulates Cdk5 activity and promotes neurodegeneration. Nature 402(6762):615–622

    Article  CAS  PubMed  Google Scholar 

  • Rashidian J, Rousseaux MW, Venderova K, Qu D, Callaghan SM, Phillips M, Bland RJ, During MJ, Mao Z, Slack RS, Park DS (2009) Essential role of cytoplasmic cdk5 and Prx2 in multiple ischemic injury models, in vivo. J Neurosci 29(40):12497–12505

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Reimann-Philipp U, Ovase R, Weigel PH, Grammas P (2001) Mechanisms of cell death in primary cortical neurons and PC12 cells. J Neurosci Res 64(6):654–660

    Article  CAS  PubMed  Google Scholar 

  • Saito T, Onuki R, Fujita Y, Kusakawa G, Ishiguro K, Bibb JA, Kishimoto T, Hisanaga S (2003) Developmental regulation of the proteolysis of the p35 cyclin-dependent kinase 5 activator by phosphorylation. J Neurosci 23(4):1189–1197

    CAS  PubMed  Google Scholar 

  • Sharma P, Veeranna SM, Amin ND, Sihag RK, Grant P, Ahn N, Kulkarni AB, Pant HC (2002) Phosphorylation of MEK1 by cdk5/p35 down-regulates the mitogen-activated protein kinase pathway. J Biol Chem 277(1):528–534

    Article  CAS  PubMed  Google Scholar 

  • Slevin M, Krupinski J (2009) Cyclin-dependent kinase-5 targeting for ischaemic stroke. Curr Opin Pharmacol 9(2):119–124

    Article  CAS  PubMed  Google Scholar 

  • Sundaram JR, Poore CP, Sulaimee NH, Pareek T, Asad AB, Rajkumar R, Cheong WF, Wenk MR, Dawe GS, Chuang KH, Pant HC, Kesavapany S (2013) Specific inhibition of p25/Cdk5 activity by the Cdk5 inhibitory peptide reduces neurodegeneration in vivo. J Neurosci 33(1):334–343

    Article  CAS  PubMed  Google Scholar 

  • Tang X, Wang X, Gong X, Tong M, Park D, Xia Z, Mao Z (2005) Cyclin-dependent kinase 5 mediates neurotoxin-induced degradation of the transcription factor myocyte enhancer factor 2. J Neurosci 25(19):4823–4834

    Article  CAS  PubMed  Google Scholar 

  • Wu J, Yang S, Xi G, Song S, Fu G, Keep RF, Hua Y (2008) Microglial activation and brain injury after intracerebral hemorrhage. Acta Neurochir Suppl 105:59–65

    Article  CAS  PubMed  Google Scholar 

  • Xue M, Del Bigio MR (2000) Intracerebral injection of autologous whole blood in rats: time course of inflammation and cell death. Neurosci Lett 283(3):230–232

    Article  CAS  PubMed  Google Scholar 

  • Xue M, Fan Y, Liu S, Zygun DA, Demchuk A, Yong VW (2009) Contributions of multiple proteases to neurotoxicity in a mouse model of intracerebral haemorrhage. Brain 132(Pt 1):26–36

    PubMed  Google Scholar 

  • Yang S, Song S, Hua Y, Nakamura T, Keep RF, Xi G (2008) Effects of thrombin on neurogenesis after intracerebral hemorrhage. Stroke 39(7):2079–2084

    Article  CAS  PubMed  Google Scholar 

  • Zhang J, Krishnamurthy PK, Johnson GV (2002) Cdk5 phosphorylates p53 and regulates its activity. J Neurochem 81(2):307–313

    Article  CAS  PubMed  Google Scholar 

  • Zhou Y, Wang Y, Wang J, Anne Stetler R, Yang QW (2014) Inflammation in intracerebral hemorrhage: from mechanisms to clinical translation. Prog Neurobiol 115:25–44

    Article  CAS  PubMed  Google Scholar 

  • Zhu J, Li W, Mao Z (2011) Cdk5: mediator of neuronal development, death and the response to DNA damage. Mech Ageing Dev 132(8–9):389–394

    Article  PubMed Central  CAS  PubMed  Google Scholar 

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Acknowledgments

This work is supported by the National Natural Science Foundation of China (81371299, 81471188, 31300902); the Colleges and Universities in Natural Science Research Project of Jiangsu Province (13KJB310009), and A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD); Technology Innovation Programme of Jiangsu province (YKC13075, YKC13086); and Technology Innovation Programme of Nantong University (CXLX13_876, CXLX13_875). The authors declare no conflict of interest.

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Correspondence to Xiaomei Chen or Yonghua Liu.

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Kaifu Ke and Jiabing Shen contributed equally to this work.

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Ke, K., Shen, J., Song, Y. et al. CDK5 Contributes to Neuronal Apoptosis via Promoting MEF2D Phosphorylation in Rat Model of Intracerebral Hemorrhage. J Mol Neurosci 56, 48–59 (2015). https://doi.org/10.1007/s12031-014-0466-5

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  • DOI: https://doi.org/10.1007/s12031-014-0466-5

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