Cellular model of neuronal atrophy induced by DYNC1I1 deficiency reveals protective roles of RAS-RAF-MEK signaling

Abstract

Neuronal atrophy is a common pathological feature occurred in aging and neurodegenerative diseases. A variety of abnormalities including motor protein malfunction and mitochondrial dysfunction contribute to the loss of neuronal architecture; however, less is known about the intracellular signaling pathways that can protect against or delay this pathogenic process. Here, we show that the DYNC1I1 deficiency, a neuron-specific dynein intermediate chain, causes neuronal atrophy in primary hippocampal neurons. With this cellular model, we are able to find that activation of RAS-RAF-MEK signaling protects against neuronal atrophy induced by DYNC1I1 deficiency, which relies on MEK-dependent autophagy in neuron. Moreover, we further reveal that BRAF also protects against neuronal atrophy induced by mitochondrial impairment. These findings demonstrate protective roles of the RAS-RAF-MEK axis against neuronal atrophy, and imply a new therapeutic target for clinical intervention.

Keywords

RAS-RAF-MEK pathway / atrophy / dynein intermediate chain / mitochondria / hippocampal neuron / autophagy

References

[1] Banks GT, Fisher EM (2008) Cytoplasmic dynein could be key to understanding neurodegeneration. Genome Biol 9:214

[2] Becker RE, Greig NH, Giacobini E (2008) Why do so many drugs for Alzheimer’s disease fail in development?Time for new methods and new practices? J Alzheimers Dis 15:303–325

[3] Boylan KLM, Hays TS (2002) The gene for the intermediate chain subunit of cytoplasmic dynein is essential in Drosophila. Genetics 162:1211–1220

[4] Brunet A, Datta SR, Greenberg ME (2001) Transcription-dependent and-independent control of neuronal survival by the PI3K–Akt signaling pathway. Curr Opin Neurobiol 11:297–305

[5] Chao MV (2003) Neurotrophins and their receptors: a convergence point for many signalling pathways. Nat Rev Neurosci 4:299–309

[6] Chen X-J, Levedakou EN, Millen KJ, Wollmann RL, Soliven B, Popko B(2007) Proprioceptive sensory neuropathyin mice witha mutation in the cytoplasmic dynein heavy chain 1 gene. J Neurosci 27:14515–14524

[7] Chin PC, Liu L, Morrison BE, Siddiq A, Ratan RR, Bottiglieri T, D’Mello SR (2004) The c-Raf inhibitor GW5074 provides neuroprotection in vitro and in an animal model of neurodegeneration through a MEK-ERK and Akt-independent mechanism. J Neurochem 90:595–608

[8] Craig LA, Hong NS, McDonald RJ (2011) Revisiting the cholinergic hypothesis in the development of Alzheimer’s disease. Neurosci Biobehav Rev 35:1397–1409

[9] Dotti CG, Sullivan CA, Banker GA (1988) The establishment of polarity byhippocampal neuronsin culture. JNeurosci 8:1454–1468

[10] Eschbach J, Dupuis L (2011) Cytoplasmic dynein in neurodegeneration. Pharmacol Ther 130:348–363

[11] Fiordalisi JJ, Johnson RL II, Ülkü AS, Der CJ, Cox AD (2001) Mammalian expression vectors for Ras family proteins: generation and use of expression constructs to analyze Ras family function. In: Der CJ, Balch WE(eds) Methods in enzymology. Academic Press, San Diego, pp3–36

[12] Fox NC, Schott JM (2004) Imaging cerebral atrophy: normal ageing to Alzheimer’s disease. Lancet 363:392–394

[13] Freeman SH, Kandel R, Cruz L, Rozkalne A, Newell K, Frosch MP, Hedley-Whyte ET, Locascio JJ, Lipsitz L, Hyman BT (2008) Preservation of neuronal number despite age-related cortical brain atrophy in elderly subjects without Alzheimer disease. J Neuropathol Exp Neurol 67:1205–1212

[14] Galabova-Kovacs G, Kolbus A, Matzen D, Meissl K, Piazzolla D, Rubiolo C, Steinitz K, Baccarini M (2006) ERK and beyond: insights from B-Raf and Raf-1 conditional knockouts. Cell Cycle Georget. Tex 5:1514–1518

[15] Ha J, Lo KW-H, Myers KR, Carr TM, Humsi MK, Rasoul BA, Segal RA, Pfister KK (2008) A neuron-specific cytoplasmic dynein isoform preferentially transports TrkB signaling endosomes. J Cell Biol 181:1027–1039

[16] Hafezparast M, Klocke R, Ruhrberg C, Marquardt A, Ahmad-Annuar A, Bowen S, Lalli G, Witherden AS, Hummerich H, Nicholson S (2003) Mutations in dynein link motor neuron degeneration to defects in retrograde transport. Science 300:808–812

[17] Heumann R, Goemans C, Bartsch D, Lingenhöhl K,Waldmeier PC, Hengerer B, Allegrini PR, Schellander K, Wagner EF, Arendt T (2000) Transgenic activation of Ras in neurons promotes hypertrophy and protects from lesion-induced degeneration. J Cell Biol 151:1537–1548

[18] Holzbaur ELF, Vallee RB (1994) Dyneins: molecular structure and cellular function. Annu Rev Cell Biol 10:339–372

[19] Kapitein LC, Schlager MA, Kuijpers M, Wulf PS, van Spronsen M, MacKintosh FC, Hoogenraad CC (2010) Mixed microtubules steer dynein-driven cargo transport into dendrites. Curr Biol 20:290–299

[20] Karran E, Mercken M, Strooper BD (2011) The amyloid cascade hypothesis for Alzheimer’s disease: an appraisal for the development of therapeutics. Nat. Rev. Drug Discov. 10:698–712

[21] Kimura S, Noda T, Yoshimori T (2008) Dynein-dependent movement of autophagosomes mediates efficient encounters with lysosomes. Cell Struct Funct 33:109–122

[22] Klionsky DJ, Abdalla FC, Abeliovich H, Abraham RT, Acevedo-Arozena A, Adeli K, Agholme L, Agnello M, Agostinis P, Aguirre-Ghiso JA (2012) Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy 8:445–544

[23] Kumar V, Zhang M-X, Swank MW, Kunz J, Wu G-Y (2005) Regulation of dendritic morphogenesis by Ras–PI3K–Akt–mTOR and Ras–MAPK signalingpathways. JNeurosci 25:11288–11299

[24] Lalli G (2014) Regulation of neuronal polarity. Exp Cell Res 328:267–275

[25] Lipka J, Kuijpers M, Jaworski J, Hoogenraad CC (2013) Mutations in cytoplasmic dynein and its regulators cause malformations of cortical development and neurodegenerative diseases. Biochem SocTrans 41:1605–1612

[26] Maday S, Wallace KE, Holzbaur ELF (2012) Autophagosomes initiate distally and mature during transport toward the cell soma in primary neurons. J Cell Biol 196:407–417

[27] Mazzoni IE, Saïd FA, Aloyz R, Miller FD, Kaplan D (1999) Ras regulates sympathetic neuron survival by suppressing the p53mediated cell death pathway. J Neurosci 19:9716–9727

[28] Mitchell DJ, Blasier KR, Jeffery ED, Ross MW, Pullikuth AK, Suo D,Park J, Smiley WR, Lo KW-H, Shabanowitz J (2012) Trk activation of the ERK1/2 kinase pathway stimulates intermediate chain phosphorylationand recruitscytoplasmicdyneintosignalingendosomes for retrograde axonaltransport. JNeurosci 32:15495–15510

[29] Moreira PI, Zhu X, Wang X, Lee H, Nunomura A, Petersen RB, Perry G, Smith MA (2010) Mitochondria: a therapeutic target in neurodegeneration. Biochim Biophys Acta 1802:212–220

[30] Myers KR, Lo KW-H, Lye RJ, Kogoy JM, Soura V, Hafezparast M, Pfister KK (2007) Intermediate chain subunit as a probe for cytoplasmic dynein function: biochemical analyses and live cell imaging in PC12 cells. J Neurosci Res 85:2640–2647

[31] Pakkenberg B, Pelvig D, Marner L, Bundgaard MJ, Gundersen HJG, Nyengaard JR, Regeur L(2003) Aging and the human neocortex. Exp Gerontol 38:95–99

[32] Payne BAI, Chinnery PF (2015) Mitochondrial dysfunction in aging: much progress but many unresolved questions. Biochim. Biophys. Acta BBA-Bioenerg. 1847:1347–1353

[33] Pfister KK, Fisher EMC, Gibbons IR, Hays TS, Holzbaur ELF, McIntosh JR, Porter ME, Schroer TA, Vaughan KT, Witman GB (2005) Cytoplasmic dynein nomenclature. J Cell Biol 171:411–413

[34] Ravikumar B, Acevedo-Arozena A, Imarisio S, Berger Z,Vacher C, O’Kane CJ, Brown SDM, Rubinsztein DC (2005) Dynein mutations impair autophagic clearance of aggregate-prone proteins. Nat Genet 37:771–776

[35] Regeur L, Badsberg Jensen G, Pakkenberg H, Evans SM, Pakkenberg B (1994) No global neocortical nerve cell loss in brains from patients with senile dementia of Alzheimer’s type. Neurobiol Aging 15:347–352

[36] Rosse C, Boeckeler K, Linch M, Radtke S, Frith D, Barnouin K, Morsi AS, Hafezparast M, Howell M, Parker PJ (2012) Binding of dynein intermediate chain 2 to paxillin controls focal adhesion dynamics and migration. J Cell Sci 125:3733–3738

[37] Scaduto RC Jr, Grotyohann LW (1999) Measurement of mitochondrial membrane potential using fluorescent rhodamine derivatives. BiophysJ 76:469–477

[38] Schott JM, Fox NC, Frost C, Scahill RI, Janssen JC, Chan D, Jenkins R, Rossor MN (2003) Assessing the onset of structural change in familial Alzheimer’s disease. Ann Neurol 53:181–188

[39] Slack C, Alic N, Foley A, Cabecinha M, Hoddinott MP, Partridge L (2015) The Ras-Erk-ETS-signaling pathway is a drug target for longevity. Cell 162:72–83

[40] Song A, Wang D, Chen G, Li Y, Luo J, Duan S, Poo M (2009) A selective filter for cytoplasmic transport at the axon initial segment. Cell 136:1148–1160

[41] Soo KY, Farg M, Atkin JD (2011) Molecular motor proteins and amyotrophic lateral sclerosis. IntJ Mol Sci 12:9057–9082

[42] Swaab DF, Hofman MA, Lucassen PJ, Salehi A, Uylings HBM (1994) Neuronal atrophy, not cell death, is the main hallmark of Alzheimer’s disease. Neurobiol Aging 15:369–371

[43] Vaughan PS, Leszyk JD, Vaughan KT (2001) Cytoplasmic dynein intermediate chain phosphorylation regulates binding to dynactin. J Biol Chem 276:26171–26179

[44] Wang J, Whiteman MW, Lian H, Wang G, Singh A, Huang D, Denmark T (2009) A non-canonical MEK/ERK signaling pathway regulates autophagy via regulating beclin 1. J Biol Chem 284:21412–21424

[45] Xie Y, Zhou B, Lin M-Y, Wang S, Foust KD, Sheng Z-H (2015) Endolysosomal deficits augment mitochondria pathology in spinal motor neurons of asymptomatic fALS mice. Neuron 87:355–370

[46] Yamaguchi T, Kakefuda R, Tajima N, Sowa Y, Sakai T (2011) Antitumor activities of JTP-74057 (GSK1120212), a novel MEK1/ 2inhibitor,on colorectalcancercell linesin vitroandin vivo. IntJ Oncol 39:23–31

[47] Zheng Y, Wildonger J, Ye B, Zhang Y, Kita A, Younger SH, Zimmerman S, Jan LY, Jan YN (2008) Dynein is required for polarized dendritic transport and uniform microtubule orientation in axons. Nat Cell Biol 10:1172–1180

[48] Zhong J, Li X, McNamee C, Chen AP, Baccarini M, Snider WD (2007) Raf kinase signaling functions in sensory neuron differentiation and axon growth in vivo. Nat Neurosci 10:598–607

RIGHTS & PERMISSIONS

The Author(s) 2016. This article is published with open access at Springerlink.com and journal.hep.com.cn