Leucine-rich repeat kinase 2 (LRRK2) 是帕金森氏症的一個致病基因，其突變可以在偶發性和遺傳性的帕金森氏症病人中發現。LRRK2突變常伴隨著α-synuclein，路易氏體 (Lewy body) 和Tau蛋白的不正常堆積。然而，在我們最近的研究中發現野生型的LRRK2在果蠅中扮演保護的角色。在這裡我們用Gal4/UAS系統，在果蠅的多巴胺神經元內同時表現野生型的人類LRRK2和野生型的人類Tau蛋白，研究兩者的關聯性。在我們的實驗中，發現在多巴胺神經元表現LRRK2能使表現Tau的果蠅壽命延長，而且其運動能力在早期也有所恢復，不僅如此，被視為是不正常的磷酸化Tau的量也因此減少。另外，我們將螢光蛋白標記在粒線體上，用於觀察粒線體在多巴胺神經元的堆積情形，我們發現LRRK2不能使Tau在PAL和PPM3不正常堆積的粒線體回復。整體而言，野生型LRRK2能夠保護Tau對果蠅造成的毒性，但不是透過回復軸突內粒線體的累積。

Leucine-rich repeat kinase 2 (LRRK2) mutations has been found to associate with sporadic and familial PD. It accompanied with abnormal protein accumulation, like α-synuclein, Lewy body and microtubule-associated protein Tau (MAPT) pathology in human. However, we recently found the protective role of wild type LRRK2 in the model organism. Here, we used UAS/Gal4 system to study the interaction between human LRRK2 and Tau. We co-expressed wild type human LRRK2 and Tau in Drosophila dopaminergic neurons. In our study, we found that wild type LRRK2 could extend Tau induced short lifespan, recovered motor disability in early age and decreased abnormal hyper-phosphorylated Tau. Additionally, we used fluorescent protein, DsRed, to label mitochondrion and observed the increase of abnormal axonal mitochondrial localization and their accumulation in dopaminergic neurons caused by Tau expression.
We found that LRRK2 barely reversed the Tau induced abnormal axonal mitochondrial accumulation in PAL and PPM3 dopaminergic neurons. In summary, human wild-type LRRK2 may protect dopaminergic neurons from Tau pathology but not through reversing axonal mitochondrial accumulation.

Baird, G. S., Zacharias, D. A., & Tsien, R. Y. (2000). Biochemistry, mutagenesis, and oligomerization of DsRed, a red fluorescent protein from coral. Proc Natl Acad Sci U S A, 97(22), 11984-11989.
Biskup, S., Moore, D. J., Celsi, F., Higashi, S., West, A. B., Andrabi, S. A., . . . Dawson, V. L. (2006). Localization of LRRK2 to membranous and vesicular structures in mammalian brain. Ann Neurol, 60(5), 557-569. doi: 10.1002/ana.21019
Biskup, S., Moore, D. J., Celsi, F., Higashi, S., West, A. B., Andrabi, S. A., . . . Dawson, V. L. (2006). Localization of LRRK2 to membranous and vesicular structures in mammalian brain. Ann Neurol, 60(5), 557-569. doi: 10.1002/ana.21019
Botella, J. A., Bayersdorfer, F., Gmeiner, F., & Schneuwly, S. (2009). Modelling Parkinson's disease in Drosophila. Neuromolecular Med, 11(4), 268-280. doi: 10.1007/s12017-009-8098-6
Damiano, M., Galvan, L., Déglon, N., & Brouillet, E. (2010). Mitochondria in Huntington's disease. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 1802(1), 52-61. doi: http://dx.doi.org/10.1016/j.bbadis.2009.07.012
de Lau, L. M. L., & Breteler, M. M. B. (2006). Epidemiology of Parkinson's disease. The Lancet Neurology, 5(6), 525-535. doi: 10.1016/s1474-4422(06)70471-9
Fahn, S. (2003) Description of Parkinson's disease as a clinical syndrome. Vol. 991. Annals of the New York Academy of Sciences (pp. 1-14).
Friggi-Grelin, F., Coulom, H., Meller, M., Gomez, D., Hirsh, J., & Birman, S. (2003). Targeted gene expression in Drosophila dopaminergic cells using regulatory sequences from tyrosine hydroxylase. Journal of Neurobiology, 54(4), 618-627. doi: 10.1002/neu.10185
Higashi, S., Biskup, S., West, A. B., Trinkaus, D., Dawson, V. L., Faull, R. L. M., . . . Emson, P. C. (2007). Localization of Parkinson’s disease-associated LRRK2 in normal and pathological human brain. Brain research, 1155, 208-219. doi: http://dx.doi.org/10.1016/j.brainres.2007.04.034
Hosoi, T., Uchiyama, M., Okumura, E., Saito, T., Ishiguro, K., Uchida, T., . . . Hisanaga, S.-i. (1995). Evidence for cdk5 as a Major Activity Phosphorylating Tau Protein in Porcine Brain Extract. The Journal of Biochemistry, 117(4), 741-749.
Hu, Y., Li, X.-C., Wang, Z.-h., Luo, Y., Zhang, X., Liu, X.-P., . . . Liu, G.-P. (2016a). Tau accumulation impairs mitophagy via increasing mitochondrial membrane potential and reducing mitochondrial Parkin.
Hu, Y., Li, X.-C., Wang, Z.-h., Luo, Y., Zhang, X., Liu, X.-P., . . . Liu, G.-P. (2016b). Tau accumulation impairs mitophagy via increasing mitochondrial membrane potential and reducing mitochondrial Parkin. oncotarget, 7(14), 17356-17368. doi: 10.18632/oncotarget.7861
Jaleel, M., Nichols, R. J., Deak, M., Campbell, David G., Gillardon, F., Knebel, A., & Alessi, Dario R. (2007a). LRRK2 phosphorylates moesin at threonine-558: characterization of how Parkinson's disease mutants affect kinase activity. Biochemical Journal, 405(2), 307-317. doi: 10.1042/bj20070209
Jaleel, M., Nichols, R J., Deak, M., Campbell, David G., Gillardon, F., Knebel, A., & Alessi, Dario R. (2007b). LRRK2 phosphorylates moesin at threonine-558: characterization of how Parkinson's disease mutants affect kinase activity. Biochemical Journal, 405(Pt 2), 307-317. doi: 10.1042/BJ20070209
Jankovic, J. (2008). Parkinson’s disease: clinical features and diagnosis. Journal of Neurology, Neurosurgery & Psychiatry, 79(4), 368-376. doi: 10.1136/jnnp.2007.131045
Jorgensen, N. D., Peng, Y., Ho, C. C. Y., Rideout, H. J., Petrey, D., Liu, P., & Dauer, W. T. (2009). The WD40 Domain Is Required for LRRK2 Neurotoxicity. PLOS ONE, 4(12), e8463. doi: 10.1371/journal.pone.0008463
Kahsai, L., & Winther, A. M. (2011). Chemical neuroanatomy of the Drosophila central complex: distribution of multiple neuropeptides in relation to neurotransmitters. J Comp Neurol, 519(2), 290-315. doi: 10.1002/cne.22520
Kawakami, F., Yabata, T., Ohta, E., Maekawa, T., Shimada, N., Suzuki, M., . . . Obata, F. (2012). LRRK2 Phosphorylates Tubulin-Associated Tau but Not the Free Molecule: LRRK2-Mediated Regulation of the Tau-Tubulin Association and Neurite Outgrowth. PLOS ONE, 7(1), e30834. doi: 10.1371/journal.pone.0030834
Kong, E. C., Woo, K., Li, H., Lebestky, T., Mayer, N., Sniffen, M. R., . . . Wolf, F. W. (2010). A Pair of Dopamine Neurons Target the D1-Like Dopamine Receptor DopR in the Central Complex to Promote Ethanol-Stimulated Locomotion in Drosophila. PLOS ONE, 5(4), e9954. doi: 10.1371/journal.pone.0009954
Kowall, N. W., & Kosik, K. S. (1987). Axonal disruption and aberrant localization of tau protein characterize the neuropil pathology of Alzheimer's disease. Ann Neurol, 22(5), 639-643. doi: 10.1002/ana.410220514
Li, X.-C., Hu, Y., Wang, Z.-h., Luo, Y., Zhang, Y., Liu, X.-P., . . . Wang, J.-Z. (2016). Human wild-type full-length tau accumulation disrupts mitochondrial dynamics and the functions via increasing mitofusins. Scientific Reports, 6, 24756. doi: 10.1038/srep24756
http://www.nature.com/articles/srep24756#supplementary-information
Lin, C.-H., Tsai, P.-I., Wu, R.-M., & Chien, C.-T. (2010a). <em>LRRK2</em> G2019S Mutation Induces Dendrite Degeneration through Mislocalization and Phosphorylation of Tau by Recruiting Autoactivated GSK3β. The Journal of Neuroscience, 30(39), 13138-13149. doi: 10.1523/jneurosci.1737-10.2010
Lin, C.-H., Tsai, P.-I., Wu, R.-M., & Chien, C.-T. (2010b). LRRK2 G2019S Mutation Induces Dendrite Degeneration through Mislocalization and Phosphorylation of Tau by Recruiting Autoactivated GSK3β. The Journal of Neuroscience, 30(39), 13138-13149. doi: 10.1523/jneurosci.1737-10.2010
Lin, C. H., Tsai, P. I., Wu, R. M., & Chien, C. T. (2010). LRRK2 G2019S mutation induces dendrite degeneration through mislocalization and phosphorylation of tau by recruiting autoactivated GSK3ss. J Neurosci, 30(39), 13138-13149. doi: 10.1523/JNEUROSCI.1737-10.2010
Liu, G., Seiler, H., Wen, A., Zars, T., Ito, K., Wolf, R., . . . Liu, L. (2006). Distinct memory traces for two visual features in the Drosophila brain. Nature, 439(7076), 551-556. doi: http://www.nature.com/nature/journal/v439/n7076/suppinfo/nature04381_S1.html
Liu, Q., Liu, S., Kodama, L., Driscoll, M. R., & Wu, M. N. (2012). Two dopaminergic neurons signal to the dorsal fan-shaped body to promote wakefulness in Drosophila. Curr Biol, 22(22), 2114-2123. doi: 10.1016/j.cub.2012.09.008
Liu, Z., Wang, X., Yu, Y., Li, X., Wang, T., Jiang, H., . . . Smith, W. W. (2008). A Drosophila model for LRRK2-linked parkinsonism. Proc Natl Acad Sci U S A, 105(7), 2693-2698. doi: 10.1073/pnas.0708452105
Maldonado, H., Ramírez, E., Utreras, E., Pando, M. E., Kettlun, A. M., Chiong, M., . . . Valenzuela, M. A. (2011). Inhibition of Cyclin-Dependent Kinase 5 but Not of Glycogen Synthase Kinase 3-β Prevents Neurite Retraction and Tau Hyperphosphorylation Caused by Secretable Products of Human T-Cell Leukemia Virus Type I-Infected Lymphocytes. Journal of Neuroscience Research, 89(9), 1489-1498. doi: 10.1002/jnr.22678
Mao, Z., & Davis, R. L. (2009). Eight different types of dopaminergic neurons innervate the Drosophila mushroom body neuropil: anatomical and physiological heterogeneity. Front Neural Circuits, 3, 5. doi: 10.3389/neuro.04.005.2009
Meixner, A., Boldt, K., Van Troys, M., Askenazi, M., Gloeckner, C. J., Bauer, M., . . . Ueffing, M. (2011). A QUICK Screen for Lrrk2 Interaction Partners – Leucine-rich Repeat Kinase 2 is Involved in Actin Cytoskeleton Dynamics. Molecular & Cellular Proteomics, 10(1). doi: 10.1074/mcp.M110.001172
Melrose, H. L., Dachsel, J. C., Behrouz, B., Lincoln, S. J., Yue, M., Hinkle, K. M., . . . Farrer, M. J. (2010). Impaired dopaminergic neurotransmission and microtubule-associated protein tau alterations in human LRRK2 transgenic mice. Neurobiol Dis, 40(3), 503-517. doi: 10.1016/j.nbd.2010.07.010
Moreira, P. I., Carvalho, C., Zhu, X., Smith, M. A., & Perry, G. (2010). Mitochondrial dysfunction is a trigger of Alzheimer's disease pathophysiology. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 1802(1), 2-10. doi: http://dx.doi.org/10.1016/j.bbadis.2009.10.006
Narendra, D., Tanaka, A., Suen, D.-F., & Youle, R. J. (2008). Parkin is recruited selectively to impaired mitochondria and promotes their autophagy. The Journal of Cell Biology, 183(5), 795-803. doi: 10.1083/jcb.200809125
Obeso, J., Rodrguez-Oroz, M., Benitez Temino, B., Blesa, F., Guridi, J., Marin, C., & Rodriguez, M. (2008). Functional organization of the basal ganglia: Therapeutic implications for Parkinson's disease. Movement disorders, 23(S3), S548-S559.
Ohta, E., Kawakami, F., Kubo, M., & Obata, F. (2011). LRRK2 directly phosphorylates Akt1 as a possible physiological substrate: Impairment of the kinase activity by Parkinson’s disease-associated mutations. FEBS Letters, 585(14), 2165-2170. doi: http://dx.doi.org/10.1016/j.febslet.2011.05.044
Ohta, E., Kawakami, F., Kubo, M., & Obata, F. (2012). LRRK2 directly phosphorylates Akt1 as a possible physiological substrate: impairment of the kinase activity by Parkinson's disease-associated mutations. Journal of neurochemistry, 123(supplement), 104-104.
Parkinson, J. (2002). An Essay on the Shaking Palsy. The Journal of Neuropsychiatry and Clinical Neurosciences, 14(2), 223-236. doi: 10.1176/jnp.14.2.223
Reddy, P. H. (2011). Abnormal Tau, Mitochondrial Dysfunction, Impaired Axonal Transport of Mitochondria, and Synaptic Deprivation in Alzheimer’s Disease. Brain research, 1415, 136-148. doi: 10.1016/j.brainres.2011.07.052
Seirafi, M., Kozlov, G., & Gehring, K. (2015). Parkin structure and function. The Febs Journal, 282(11), 2076-2088. doi: 10.1111/febs.13249
Shanley, M. R., Hawley, D., Leung, S., Zaidi, N. F., Dave, R., Schlosser, K. A., . . . Liu, M. (2015). LRRK2 Facilitates tau Phosphorylation through Strong Interaction with tau and cdk5. Biochemistry, 54(33), 5198-5208. doi: 10.1021/acs.biochem.5b00326
Sperbera, B. R., Leight, S., Goedert, M., & Lee, V. M. Y. (1995). Glycogen synthase kinase-3β phosphorylates tau protein at multiple sites in intact cells. Neuroscience Letters, 197(2), 149-153. doi: https://doi.org/10.1016/0304-3940(95)11902-9
Steger, M., Tonelli, F., Ito, G., Davies, P., Trost, M., Vetter, M., . . . Mann, M. (2016). Phosphoproteomics reveals that Parkinson's disease kinase LRRK2 regulates a subset of Rab GTPases. eLife, 5, e12813. doi: 10.7554/eLife.12813
Strauss, R. (2002). The central complex and the genetic dissection of locomotor behaviour. Current Opinion in Neurobiology, 12(6), 633-638. doi: https://doi.org/10.1016/S0959-4388(02)00385-9
Thomas, J. M., Li, T., Yang, W., Xue, F., Fishman, P. S., & Smith, W. W. (2017). 68 and FX2149 Attenuate Mutant LRRK2-R1441C-Induced Neural Transport Impairment. Frontiers in Aging Neuroscience, 8(337). doi: 10.3389/fnagi.2016.00337
Wang, X., Yan, M. H., Fujioka, H., Liu, J., Wilson-Delfosse, A., Chen, S. G., . . . Zhu, X. (2012). LRRK2 regulates mitochondrial dynamics and function through direct interaction with DLP1. Hum Mol Genet, 21(9), 1931-1944. doi: 10.1093/hmg/dds003
Wang, Y., & Mandelkow, E. (2016). Tau in physiology and pathology. Nat Rev Neurosci, 17(1), 22-35. doi: 10.1038/nrn.2015.1
Winklhofer, K. F., & Haass, C. (2010). Mitochondrial dysfunction in Parkinson's disease. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 1802(1), 29-44. doi: http://dx.doi.org/10.1016/j.bbadis.2009.08.013
Wu, T.-H., Lu, Y.-N., Chuang, C.-L., Wu, C.-L., Chiang, A.-S., Krantz, D. E., & Chang, H.-Y. (2013). Loss of vesicular dopamine release precedes tauopathy in degenerative dopaminergic neurons in a Drosophila model expressing human tau. Acta Neuropathologica, 125(5), 711-725. doi: 10.1007/s00401-013-1105-x
Xiong, Y., Dawson, V. L., & Dawson, T. M. (2012). LRRK2 GTPase Dysfunction in the Pathogenesis of Parkinson’s disease. Biochemical Society transactions, 40(5), 1074-1079. doi: 10.1042/BST20120093
Yun, H. J., Kim, H., Ga, I., Oh, H., Ho, D. H., Kim, J., . . . Seol, W. (2015). An early endosome regulator, Rab5b, is an LRRK2 kinase substrate. The Journal of Biochemistry, 157(6), 485-495. doi: 10.1093/jb/mvv005
Zhu, Y., Wang, C., Yu, M., Cui, J., Liu, L., & Xu, Z. (2013). ULK1 and JNK are involved in mitophagy incurred by LRRK2 G2019S expression. Protein & Cell, 4(9), 711-721. doi: 10.1007/s13238-013-3910-3