93
A40
Veh 1 mg/kg 3 mg/kg
0.0 2.5 5.0 7.5 10.0
n.s.
n.s.
A40 [pmol/g tissue]
A42
Veh 1 mg/kg 3 mg/kg 0
1 2 3 4 5 A42 [fmol/g tissue]
n.s.
n.s.
A40
Veh 1 mg/kg 3 mg/kg
0 25 50 75 100
n.s.
n.s.
A40 [pmol/g tissue]
A42
Veh 1 mg/kg 3 mg/kg 0
100 200 300 400 500 n.s.
n.s.
A42 [pmol/g tissue]
pT688-APP
94
Figure16. Behavioral improvement in the Y-maze after MMBO administration in 3xTg-AD mice. MMBO (1 and 3 mg/kg, b.i.d) or vehicle was administered for 16 days, 30 minutes prior to the Y-maze test. Data are presented as % alternation rate (A) and total frequency of arm entries (B) during an 8-minute exploration. Data were expressed as mean ± SEM and analyzed statistically as follows: Student’s t-test (WT vs. Tg, * p ≤ 0.05,
** p ≤ 0.01) and one-tailed Williams’ test (vs. vehicle, # p ≤ 0.025), n = 10.
95
A B
Spontaneous alternation
Veh Veh 1 mg/kg 3 mg/kg 0
10 20 30 40 50 60 70 80 90
3xTg-AD
**
##
WT
% alternation
Total entries
Veh Veh 1 mg/kg 3 mg/kg 0
5 10 15 20 25
3xTg-AD
*
WT
Frequency
96
Figure17. Behavioral improvement in a novel object recognition test after MMBO administration in 3xTg-AD mice. MMBO (1 and 3 mg/kg, b.i.d) or vehicle was administered for 24 days, 30 minutes prior to the acquisition phase. Exploration of a novel object was evaluated during retention phase, 5 hours after acquisition. Novel object recognition performance is shown as exploration frequency (A) or novel objectpreference ratio (B). Data were expressed as mean ± SEM and analyzed statistically as follows: Student’s t-test (novel object vs. familiar object or WT vs. Tg, * p
≤ 0.05,** p ≤ 0.01, *** p ≤ 0.001) and one-tailed Williams’ test (vs. vehicle, # p ≤ 0.025), n
= 10.
97
Veh Veh 1 mg/kg 3 mg/kg 0
5 10
15 Object A (familiar)
Object B (novel)
3xTg-AD n.s.
***
* *
WT
Exploration frequency
Veh Veh 1 mg/kg 3 mg/kg 0
25 50 75 100
3xTg-AD
# #
***
Recognition Index [%]
WT
A B
98
Supplementary Figure8. Amounts of tau in JNPL3 and 3xTg-AD mice. Total mouse and human tau, total human tau, and tau phosphorylated at T205, S396, T181, and S202/T205 were determined by immunoblot analysis (A). Quantitative comparisons of JNPL3 and 3xTg-AD mice are shown respectively in B-G. Data are expressed as scatter plots, n = 7 or 8.
99
Ab-3 (total m&h tau)
WT Tg WT Tg
0 1000000 2000000 3000000 4000000 5000000 6000000
3xTg-AD JNPL3
AU
HT7 (total htau)
WT Tg WT Tg
0 1000000 2000000 3000000 4000000 5000000 6000000
3xTg-AD JNPL3
AU
pT205 tau
WT Tg WT Tg
0 1000000 2000000 3000000 4000000 5000000
3xTg-AD JNPL3
AU
AT270 (pT181 tau)
WT Tg WT Tg
0 1000000 2000000 3000000
3xTg-AD JNPL3
AU
AT8
WT Tg WT Tg
0 1000000 2000000 3000000 4000000
3xTg-AD JNPL3
AU
pS396 tau
WT Tg WT Tg
0 500000 1000000 1500000 2000000
3xTg-AD JNPL3
AU
Ab-3 (total tau)
AT270 (pT181) pT205 AT8 (pS202/pT205) pS396 HT7 (htau)
WT WT
JNPL3 3xTg-AD
A B C
D E
F G
100
General Discussion
101
In this study, I tried to examine the relationship between tau and etiology in neurodegenerative disease such as AD by generating a novel Tg mouse model (part 1), and I also tested potential of tau-targeting drug, which has been originally discovered (part 2). Tau, a MT protein is a key molecule in neurodegenerative diseases because it is accumulated as NFTs in a diseased brain and mutations in tau gene is known to cause a familial type of neurodegenerative disease (Hutton et al., 1998; Poorkaj et al., 1998;
Spillantini et al., 1998). Toxicity of tau is not completely understood yet, however, hyperphosphorylation or mutations of tau is known to reduce the binding to MT through conformation change (Grundke-Iqbal et al., 1986; Goedert et al., 1995; Dan and Hasegawa., 2011). Detachment of tau from MT could destabilize MT and impair axonal function. Increased soluble pool of tau with mutation or hyperphosphorylation is prone to aggregate and it would damage neurons. Namely, mislocalization of tau might a key event in neurodegenerative diseases.
In part 1 of this study, a novel mutant tau Tg mouse TPR50 was generated and evaluated biochemically and behaviorally to analyze tau-induced neurotoxicity. TPR50 mouse dramatically displayed neurodegenerative disease-like phenotype such as age-dependent tau aggregation, motor dysfunction, and shortened life expectancy (Figures 1, 4 and 5, and Supplementary Figure 2). Additionally, impaired axonal transport as well as behavioral abnormality was clearly observed at early onset (Figures 6 and 9). Interestingly, as axonal dysfunction precedes obvious appearance of NFT-like pathology, increase in soluble and phosphorylated tau may perturb axonal function.
Actually, it has been recently emphasized that soluble tau like oligomers plays an important role in neuronal dysfunction (Wittmann et al., 2001; SantaCruz et al., 2005;
Cárdenas-Aguayo Mdel et al., 2014). Although molecular mechanism underlying axonal
102
impairment in TPR50 is still unknown, overexpressed mutant tau may interfere in function of endogenous tau and induce axonal dysfunction. In this study, MT hyperdynamism and increase in MT-related proteins were observed (Figures 7 and 8).
Those observations might reflect imbalanced turnover of MT due to overload of mutant tau. Based on this result, it is hypothesized that targeting mislocalization of tau could be a promising therapeutic approach via reducing intracellular soluble tau oligomers and rescuing axonal function.
As one of candidate therapeutic approaches, inhibition of tau phosphorylation was focused in part 2. I discovered a novel inhibitor for GSK-3, a major kinase for tau, by efforts in the high-throughput screening and chemistry modification, and named MMBO. MMBO displays good selectivity for GSK-3, bioavailability, and brain permeability (Saitoh et al., 2009b and Table 2). Moreover, MMBO significantly showed therapeutic potential with reduction of tau phosphorylation and behavioral amelioration in AD mouse model (Figures 13 and 16), suggesting inhibition of tau phosphorylation would be a valid therapeutic strategy for AD as per the hypothesis.
Regarding tau accumulation, MMBO did not change HT7-positive (total tau) pathology significantly (Figure 14). Therefore, observed behavioral amelioration could be dependent on reduction of soluble phosphorylated tau rather than effect of tau aggregate. This is consistent with my observation in TPR50 in part 1. In this study, I did not examine effect of MMBO on axonal transport or MT function. However, recent paper indicated that overexpression of tau with phosphomimetic mutations impaired axonal transport of mitochondria greater than WT tau (Shahpasand et al., 2012), suggesting that inhibition of tau phosphorylation might improve axonal function. Interestingly, pathological tau can enhance KLC phosphorylation by GSK-3 through activation of PP1
103
and following GSK-3 activation. KLC phosphorylation dissociates kinesin from cargo vesicles and impairs kinesin-dependent axonal transport (LaPointe et al., 2009; Kanaan et al., 2011). Namely, GSK-3 activation and consequent tau phosphorylation could lead to a vicious circle on axonal function. While, regarding dynein-dependent retrograde transport, tau accumulation might also impair it by affecting dynein/dynactin complex as reported by the recent study using flies (Butzlaff et al., 2015). Future study using MMBO will clarify relationship between tau phosphorylation and axonal function and the mechanism of action in its therapeutic effect.
In conclusion, improvement of axonal dysfunction due to abnormal tau is a novel therapeutic concept in neurodegenerative diseases such as AD, and inhibition of tau phosphorylation is a promising approach based on the concept.
104
Acknowledgements
105
I am most grateful to Professors Osamu Numata and Kazuto Nakata, and Associate Professors Kentaro Nakano and Kazuichi Sakamoto, University of Tsukuba, for their continuous guidance and valuable discussions through my doctoral program.
I thank Dr. Frank M. LaFerla for providing 3xTg-AD mice, KineMed, Inc. and Mr. Yoji Ueda for measurement of MT dynamics, Drs. Keisuke Hirai, Shinichi Kondou, Ken-ichi Noguchi, Tadatoshi Hashimoto, Hideaki Nagaya, and Takeo Wada for their encouragement, and Mr. Ryota Maeda, Ms. Yumiko Uno, Mr. Shunya Suzuki, Dr. Hideki Matsui, Mr. Masashi Yamaguchi, Dr. Keiji Yamamoto, Dr. Tomohiro Kawamoto, and Dr.
Hideki Takahashi for helpful discussions.
106 References
107
Allen B, Ingram E, Takao M., Smith MJ, Jakes R, Virdee K, Yoshida H, Holzer M., Craxton M, Emson PC, Atzori C, Migheli A, Crowther RA, Ghetti B, Spillantini MG, Goedert M. (2002). Abundant tau filaments and nonapoptotic neurodegeneration in transgenic mice expressing human P301S tau protein. J. Neurosci. 22, 9340‒9351.
Alzheimer's Disease International: World Alzheimer Report 2015, Available in http://www.alz.co.uk/research/world-report-2015
alz.org: alzheimer's association a (http://www.alz.org/dementia/types-of-dementia.asp) alz.org: alzheimer's association b (http://www.alz.org/jp/dementia-alzheimers-japan.asp) Asami-Odaka A, Ishibashi Y, Kikuchi T, Kitada C, Suzuki N. (1995). Long amyloid beta-protein secreted from wild-type human neuroblastoma IMR-32 cells. Biochemistry 34, 10272-10278.
Aplin AE, Gibb GM, Jacobsen JS, Gallo JM, Anderton BH. (1996). In vitro phosphorylation of the cytoplasmic domain of the amyloid precursor protein by glycogen synthase kinase-3beta. J Neurochem. 67, 699-707.
Barten DM, Fanara P, Andorfer C, Hoque N, Wong PY, Husted KH, Cadelina GW, Decarr LB, Yang L, Liu V, Fessler C, Protassio J, Riff T, Turner H, Janus CG, Sankaranarayanan S, Polson C, Meredith JE, Gray G, Hanna A, Olson RE, Kim SH, Vite GD, Lee FY, Albright CF. (2012). Hyperdynamic microtubules, cognitive deficits, and pathology are improved in tau transgenic mice with low doses of the microtubule-stabilizing agent BMS-241027. J. Neurosci. 32, 7137‒7145.
Belarbi K, Schindowski K, Burnouf S, Caillierez R, Grosjean ME, Demeyer D, Hamdane, M, Sergeant N, Blum D, Buée L. (2009). Early Tau pathology involving the septo-hippocampal pathway in a Tau transgenic model: relevance to Alzheimer's disease.
Curr Alzheimer Res. 6, 152‒157.
108
Belarbi K, Burnouf S, Fernandez-Gomez FJ, Desmercières J, Troquier L, Brouillette J, Tsambou L, Grosjean ME, Caillierez R, Demeyer D, Hamdane M., Schindowski K, Blum D, Buée L. (2011). Loss of medial septum cholinergic neurons in THY-Tau22 mouse model: what links with tau pathology? Curr Alzheimer Res. 8, 633‒638.
Bellucci A, Luccarini I, Scali C, Prosperi C, Giovannini MG, Pepeu G, Casamenti F.
(2006). Cholinergic dysfunction, neuronal damage and axonal loss in TgCRND8 mice.
Neurobiol Dis. 23, 260‒272.
Billings LM, Oddo S, Green KN, McGaugh JL. and LaFerla FM. (2005). Intraneuronal Abeta causes the onset of early Alzheimer's disease-related cognitive deficits in transgenic mice. Neuron 45, 675-688.
Black MM, Baas PW, Humphries S. (1989). Dynamics of alpha-tubulin deacetylation in intact neurons. J. Neurosci. 9, 358‒368.
Brunden KR, Zhang B, Carroll J, Yao Y, Potuzak JS, Hogan AM, Iba M, James MJ, Xie S, Ballatore C, Smith AB. 3rd, Lee VM, Trojanowski JQ. (2010). Epothilone D improves microtubule density, axonal integrity, and cognition in a transgenic mouse model of tauopathy. J. Neurosci. 30, 13861‒13866.
Bugiani O, Murrell JR, Giaccone G, Hasegawa M, Ghigo S, Tabaton M, Morbin M, Primavera A, Carella F, Solaro C, Grisoli M, Savoiardo M, Spillantini MG, Tagliavini F, Goedert M, Ghetti B. (1999). Frontotemporal dementia and corticobasal degeneration in a family with a P301S mutation in Tau. J Neuropathol Exp Neurol. 58, 667‒677
Bullock BP. and Habener JF. (1998). Phosphorylation of the cAMP response element binding protein CREB by cAMP-dependent protein kinase A and glycogen synthase kinase-3 alters DNA-binding affinity, conformation, and increases net charge.
Biochemistry 37, 3795-3809.
109
Butzlaff M, Hannan SB, Karsten P, Lenz S, Ng J, Voßfeldt H, Prüßing K, Pflanz R, Schulz JB, Rasse T, Voigt A. (2015). Impaired retrograde transport by the Dynein/Dynactin complex contributes to Tau-induced toxicity. Hum Mol Genet. 24, 3623-3637
Caccamo A, Oddo S, Tran LX, and LaFerla FM. (2007). Lithium reduces tau phosphorylation but not A beta or working memory deficits in a transgenic model with both plaques and tangles. Am. J. Pathol. 170, 1669-1675.
Cárdenas-Aguayo Mdel C, Gómez-Virgilio L, DeRosa S, Meraz-Ríos MA (2014). The role of tau oligomers in the onset of Alzheimer's disease neuropathology. ACS Chem Neurosci. 5, 1178-91.
Chalecka-Franaszek E. and Chuang DM. (1999). Lithium activates the serine/threonine kinase Akt-1 and suppresses glutamate-induced inhibition of Akt-1 activity in neurons.
Proc. Natl. Acad. Sci. USA 96, 8745-8750.
Clinton LK, Billings LM, Green KN, Caccamo A, Ngo J, Oddo S, McGaugh JL, and LaFerla FM. (2007). Age-dependent sexual dimorphism in cognition and stress response in the 3xTg-AD mice. Neurobiol. Dis. 28, 76-82.
Cole AR, Knebel A, Morrice NA, Robertson LA, Irving AJ, Connolly CN, and Sutherland C. (2004). GSK-3 phosphorylation of the Alzheimer epitope within collapsin response mediator proteins regulates axon elongation in primary neurons. J Biol Chem. 279, 50176-50180.
Cruz JC, and Tsai LH. (2004). A Jekyll and Hyde kinase: roles for Cdk5 in brain development and disease. Curr. Opin. Neurobiol. 14, 390-394.
Dan A, and Hasegawa M. (2011). Molecular biology of FTDP-17 (frontotemporal dementia and parkinsonism linked to chromosome 17). Nihon Rinsho. 69 Suppl 10(Pt 2),
110 379-83.
Davies SP, Reddy H, Caivano Mm and Cohen P. (2000) Specificity and mechanism of action of some commonly used protein kinase inhibitors. Biochem. J. 351, 95-105.
Dixit R, Ross JL, Goldman YE, and Holzbaur EL. (2008). Differential regulation of dynein and kinesin motor proteins by tau. Science. 319, 1086‒1089.
Drechsel DN, Hyman AA, Cobb MH, and Kirschner MW. (1992). Modulation of the dynamic instability of tubulin assembly by the microtubule-associated protein tau. Mol Biol Cell. 3, 1141‒1154.
Eckermann K, Mocanu MM, Khlistunova I, Biernat J, Nissen A, Hofmann A, Schönig K, Bujard H, Haemisch A, Mandelkow E, Zhou L, Rune G, and Mandelkow EM. (2007).
The beta-propensity of Tau determines aggregation and synaptic loss in inducible mouse models of tauopathy. J. Biol. Chem. 282, 31755–31765.
Engel T, Hernandez F, Avila J, and Lucas JJ. (2006). Full reversal of Alzheimer's disease-like phenotype in a mouse model with conditional overexpression of glycogen synthase kinase-3. J. Neurosci. 26, 5083-5090.
Fanara P, Banerjee J, Hueck RV, Harper MR, Awada M, Turner H, Husted KH, Brandt R, and Hellerstein MK. (2007). Stabilization of hyperdynamic microtubules is neuroprotective in amyotrophic lateral sclerosis. J. Biol. Chem. 282, 23465‒23472.
Fanara P, Husted KH, Selle K, Wong PY, Banerjee J, Brandt R, and Hellerstein MK.
(2010). Changes in microtubule turnover accompany synaptic plasticity and memory formation in response to contextual fear conditioning in mice. Neuroscience. 168, 167‒
178.
Fanara P, Wong PY, Husted KH, Liu S, Liu VM, Kohlstaedt LA, Riiff T, Protasio JC, Boban D, Killion S, Killian M, Epling L, Sinclair E, Peterson J, Price RW, Cabin DE,
111
Nussbaum RL, Brühmann J, Brandt R, Christine CW, Aminoff MJ, and Hellerstein MK.
(2012). Cerebrospinal fluid-based kinetic biomarkers of axonal transport in monitoring neurodegeneration. J Clin. Invest. 122, 3159‒3169.
Flunkert S, Hierzer M, Löffler T, Rabl R, Neddens J, Duller S, Schofield EL, Ward MA, Posch M, Jungwirth H, Windisch M, and Hutter-Paier B. (2013). Elevated Levels of Soluble Total and Hyperphosphorylated Tau Result in Early Behavioral Deficits and Distinct Changes in Brain Pathology in a New Tau Transgenic Mouse Model.
Neurodegener. Dis. 11, 194‒205
Friedhoff P, von Bergen M, Mandelkow EM, and Mandelkow E. (2000). Structure of tau protein and assembly into paired helical filaments. Biochim. Biophys. Acta. 1502, 122-132.
Gilman S, Koller M, Black RS, Jenkins L, Griffith SG, Fox NC, Eisner L, Kirby L, Rovira MB, Forette F, and Orgogozo JM; AN1792(QS-21)-201 Study Team. (2005).
Clinical effects of Abeta immunization (AN1792) in patients with AD in an interrupted trial. Neurology 64, 1553-1562.
Gimenez-Llort L, Blazquez G, Canete T, Johansson B, Oddo S, Tobena A, LaFerla FM, and Fernandez-Teruel A. (2007). Modeling behavioral and neuronal symptoms of Alzheimer's disease in mice: a role for intraneuronal amyloid. Neurosci. Biobehav. Rev.
31, 125-147.
Goedert M, Wischikm CM, Crowther RA, Walker JE, and Klug A. (1988). Cloning and sequencing of the cDNA encoding a core protein of the paired helical filament of Alzheimer disease: identification as the microtubule-associated protein tau. Proc Natl Acad Sci U S A. 85, 4051‒4055.
Goedert M, Spillantini MG, Jakes R, Crowther RA, Vanmechelen E, Probst A, Götz J,
112
Bürki K, and Cohen P. (1995). Molecular dissection of the paired helical filament.
Neurobiol Aging. 16, 325‒334.
Goedert M, Jakes R, and Crowther RA. (1999). Effects of frontotemporal dementia FTDP-17 mutations on heparin-induced assembly of tau filaments. FEBS. Lett. 450, 306‒311.
Golde TE, Schneider LS, and Koo EH. (2011). Anti-Aβ therapeutics in Alzheimer's disease: the need for a paradigm shift. Neuron. 2011 69 , 203-213.
Götz J, and Ittner LM. (2008). Animal models of Alzheimer's disease and frontotemporal dementia. Nat Rev Neurosci. 9, 532‒544.
Grundke-Iqbal I, Iqbal K, Tung YC, Quinlan M, Wisniewski HM, and Binder LI. (1986).
Abnormal phosphorylation of the microtubule-associated protein tau in Alzheimer cytoskeletal pathology. Proc. Natl. Acad. Sci. USA 83, 4913-4917.
Hampel H, Ewers M, Bürger K, Annas P, Mörtberg A, Bogstedt A, Frölich L, Schröder J, Schönknecht P, Riepe MW, Kraft I, Gasser T, Leyhe T, Möller HJ, Kurz A, and Basun H.
(2009). Lithium trial in Alzheimer's disease: a randomized, single-blind, placebo-controlled, multicenter 10-week study. J Clin Psychiatry. 70, 922-31.
Hansen TO, Rehfeld JF, and Nielsen FC. (2004). GSK-3beta reduces cAMP-induced cholecystokinin gene expression by inhibiting CREB binding. Neuroreport 15, 841-845.
Henthorn KS, Roux MS, Herrera C, and Goldstein LS. (2011). A role for kinesin heavy chain in controlling vesicle transport into dendrites in Drosophila. Mol Biol Cell. 22, 4038‒4046.
Hernández F, Borrell J, Guaza C, Avila J, and Lucas JJ. (2002). Spatial learning deficit in transgenic mice that conditionally over-express GSK-3beta in the brain but do not form tau filaments. J Neurochem. 83, 1529-1533.
113
Hooper C, Markevich V, Plattner F, Killick R, Schofield E, Engel T, Hernandez F, Anderton B, Rosenblum K, Bliss T, Cooke SF, Avila J, Lucas JJ, Giese KP, Stephenson J, and Lovestone S. (2007) Glycogen synthase kinase-3 inhibition is integral to long-term potentiation. Eur J Neurosci. 25, 81-86.
Hurd DD, and Saxton WM. (1996). Kinesin mutations cause motor neuron disease phenotypes by disrupting fast axonal transport in Drosophila. Genetics. 144, 1075‒
1085.
Hutton M, Lendon CL, Rizzu P, Baker M, Froelich S, Houlden H, Pickering-Brown S, Chakraverty S, Isaacs A, Grover A, Hackett J, Adamson J, Lincoln S, Dickson D, Davies P, Petersen RC, Stevens M, de Graaff E, Wauters E, van Baren J, Hillebrand M, Joosse M, Kwon JM, Nowotny P, Che LK, Norton J, Morris JC, Reed LA, Trojanowski J, Basun H, Lannfelt L, Neystat M, Fahn S, Dark F, Tannenberg T, Dodd PR, Hayward N, Kwok JB, Schofield PR, Andreadis A, Snowden J, Craufurd D, Neary D, Owen F, Oostra BA, Hardy J, Goate A, van Swieten J, Mann D, Lynch T, and Heutink P. (1998). Association of missense and 5’-splice-site mutations in tau with the inherited dementia FTDP-17.
Nature 393, 702‒705.
Iritani S, Niizato K, and Emson PC. (2001). Relationship of calbindin D28K-immunoreactive cells and neuropathological changes in the hippocampal formation of Alzheimer's disease. Neuropathology. 21, 162‒167.
Ishihara T, Hong M, Zhang B, Nakagawa Y, Lee MK, Trojanowski JQ, and Lee VM.
(1999). Age-dependent emergence and progression of a tauopathy in transgenic mice overexpressing the shortest human tau isoform. Neuron 3, 751–762.
Ishihara T, Zhang B, Higuchi M, Yoshiyama Y, Trojanowski JQ, and Lee VM. (2001).
Age-dependent induction of congophilic neurofibrillary tau inclusions in tau transgenic
114 mice. Am. J. Pathol. 158, 555–562.
Ishizawa T, Sahara N, Ishiguro K, Kersh J, McGowan E, Lewis J, Hutton M, Dickson DW, and Yen SH. (2003). Co-localization of glycogen synthase kinase-3 with neurofibrillary tangles and granulovacuolar degeneration in transgenic mice. Am. J.
Pathol. 163, 1057-1067.
Ittner LM, Fath T, Ke YD, Bi M, van Eersel J, Li KM, Gunning P, and Götz J. (2008).
Parkinsonism and impaired axonal transport in a mouse model of frontotemporal dementia. Proc. Natl. Acad. Sci. U S A. 105, 15997‒16002.
Kanaan NM, Morfini GA, LaPointe NE, Pigino GF, Patterson KR, Song Y, Andreadis A, Fu Y, Brady ST, and Binder LI. (2011). Pathogenic forms of tau inhibit kinesin-dependent axonal transport through a mechanism involving activation of axonal phosphotransferases. J. Neurosci. 31, 9858–9868.
Kidd M. (1963). Paired helical filaments in electron microscopy of Alzheimer’s disease.
Nature 197, 192–193.
Kidd M. (1964). Alzheimer’s disease: An electron microscopy study. Brain 87, 307–320.
Klein PS, and Melton DA. (1996) A molecular mechanism for the effect of lithium on development. Proc. Natl. Acad. Sci. USA 93, 8455-8459.
LaPointe NE, Morfini G, Pigino G, Gaisina IN, Kozikowski AP, Binder LI, Brady ST.
(2009). The amino terminus of tau inhibits kinesin-dependent axonal transport:
implications for filament toxicity. J. Neurosci. Res. 87, 440–451.
Le Corre S, Klafki HW, Plesnila N, Hübinger G, Obermeier A, Sahagún H, Monse B, Seneci P, Lewis J, Eriksen J, Zehr C, Yue M, McGowan E, Dickson DW, Hutton M, and Roder HM. (2006) An inhibitor of tau hyperphosphorylation prevents severe motor impairments in tau transgenic mice. Proc. Natl. Acad. Sci. USA 103, 9673-9678.
115
Lee G, Cowan N, and Kirschner M. (1998). The primary structure and heterogeneity of tau protein from mouse brain. Science. 239, 285‒288.
Lee VM, Goedert M, and Trojanowski JQ. (2001). Neurodegenerative tauopathies. Annu Rev Neurosci. 24, 1121‒1159.
Lewis J, McGowan E, Rockwood J, Melrose H, Nacharaju P, Van Slegtenhorst M, Gwinn-Hardy K, Paul Murphy M, Baker M, Yu X, Duff K, Hardy J, Corral A, Lin WL, Yen SH, Dickson DW, Davies P, and Hutton M. (2000). Neurofibrillary tangles, amyotrophy and progressive motor disturbance in mice expressing mutant (P301L) tau protein. Nat. Genet. 25, 402-405.
Liu F, Iqbal K, Grundke-Iqbal I, and Gong CX. (2002). Involvement of aberrant glycosylation in phosphorylation of tau by cdk5 and GSK-3beta. FEBS Lett. 530, 209-214.
Lossos A, Reches A, Gal A, Newman JP, Soffer D, Gomori JM, Boher M, Ekstein D, Biran I, Meiner Z, Abramsky O, and Rosenmann H. (2003). Frontotemporal dementia and parkinsonism with the P301S tau gene mutation in a Jewish family. J. Neurol. 250, 733‒740.
Lovestone S, Hartley CL, Pearce J, and Anderton BH. (1996). Phosphorylation of tau by glycogen synthase kinase-3 beta in intact mammalian cells: the effects on the organization and stability of microtubules. Neuroscience 73, 1145-1157.
Lucas JJ, Hernandez F, Gomez-Ramos P, Moran MA, Hen R, and Avila J. (2001).
Decreased nuclear beta-catenin, tau hyperphosphorylation and neurodegeneration in GSK-3beta conditional transgenic mice. EMBO J. 20, 27-39.
Murray ME, Lowe VJ, Graff-Radford NR, Liesinger AM, Cannon A, Przybelski SA, Rawal B, Parisi JE, Petersen RC, Kantarci K, Ross OA, Duara R, Knopman DS, Jack
116
CR Jr, and Dickson DW. (2015). Clinicopathologic and 11C-Pittsburgh compound B implications of Thal amyloid phase across the Alzheimer's disease spectrum. Brain. 138 (Pt 5), 1370-81.
Noble W, Olm V, Takata K, Casey E, Mary O, Meyerson J, Gaynor K, LaFrancois J, Wang L, Kondo T, Davies P, Burns M, Veeranna, Nixon R, Dickson D, Matsuoka Y, Ahlijanian M, Lau LF, and Duff K. (2003). Cdk5 is a key factor in tau aggregation and tangle formation in vivo. Neuron 38, 555-565.
Noble W, Planel E, Zehr C, Olm V, Meyerson J, Suleman F, Gaynor K, Wang L, LaFrancois J, Feinstein B, Burns M, Krishnamurthy P, Wen Y, Bhat R, Lewis J, Dickson D, and Duff K. (2005). Inhibition of glycogen synthase kinase-3 by lithium correlates with reduced tauopathy and degeneration in vivo. Proc. Natl. Acad. Sci. USA.
102, 6990-6995.
Oddo S, Caccamo A, Shepherd JD, Murphy MP, Golde TE, Kayed R, Metherate R, Mattson MP, Akbari Y, and LaFerla FM. (2003). Triple-transgenic model of Alzheimer's disease with plaques and tangles: intracellular Abeta and synaptic dysfunction. Neuron 39, 409-421.
Oddo S, Vasilevko V, Caccamo A, Kitazawa M, Cribbs DH, and LaFerla FM. (2006).
Reduction of soluble Abeta and tau, but not soluble Abeta alone, ameliorates cognitive decline in transgenic mice with plaques and tangles. J. Biol. Chem. 281, 39413-39423.
Onishi T, Iwashita H, Uno Y, Kunitomo J, Saitoh M, Kimura E, Fujita H, Uchiyama N, Kori M, and Takizawa M. (2011). A novel glycogen synthase kinase-3 inhibitor 2-methyl-5-(3-{4-[(S )-methylsulfinyl]phenyl}-1-benzofuran-5-yl)-1,3,4-oxadiazole
decreases tau phosphorylation and ameliorates cognitive deficits in a transgenic model of Alzheimer’s disease. J. Neurochem. 119, 1330‒1340.
117
Palop JJ, Chin J, Roberson ED, Wang J, Thwin MT, Bien-Ly N, Yoo J, Ho KO, Yu GQ, Kreitzer A, Finkbeiner S, Noebels JL, and Mucke L. (2007). Aberrant excitatory neuronal activity and compensatory remodeling of inhibitory hippocampal circuits in mouse models of Alzheimer's disease. Neuron 55, 697-711.
Pei JJ, Tanaka T, Tung YC, Braak E, Iqbal K, and Grundke-Iqbal I. (1997). Distribution, levels, and activity of glycogen synthase kinase-3 in the Alzheimer disease brain. J.
Neuropathol. Exp. Neurol. 56, 70-78.
Pei JJ, Braak E, Braak H, Grundke-Iqbal I, Iqbal K, Winblad B, and Cowburn RF.
(1999). Distribution of active glycogen synthase kinase 3beta (GSK-3beta) in brains staged for Alzheimer disease neurofibrillary changes. J. Neuropathol. Exp. Neurol. 58, 1010-1019.
Peineau S, Taghibiglou C, Bradley C, Wong TP, Liu L, Lu J, Lo E, Wu D, Saule E, Bouschet T, Matthews P, Isaac JT, Bortolotto ZA, Wang YT, and Collingridge GL. (2007).
LTP inhibits LTD in the hippocampus via regulation of GSK3beta. Neuron 53, 703-717.
Pérez M, Hernández F, Lim F, Díaz-Nido J, and Avila J. (2003). Chronic lithium treatment decreases mutant tau protein aggregation in a transgenic mouse model. J.
Alzheimers. Dis. 5, 301-308.
Phiel CJ, and Klein PS. (2001). Molecular targets of lithium action. Annu. Rev.
Pharmacol. Toxicol. 41, 789-813.
Phiel CJ, Wilson CA, Lee VM, and Klein PS. (2003). GSK-3alpha regulates production of Alzheimer's disease amyloid-beta peptides. Nature 423, 435-439.
Poorkaj P, Bird TD, Wijsman E, Nemens E, Garruto RM, Anderson L, Andreadis A, Wiederholt WC, Raskind M, and Schellenberg GD. (1998). Tau is a candidate gene for chromosome 17 frontotemporal dementia. Ann. Neurol. 43, 815‒825.
118
Riascos D, de Leon D, Baker-Nigh A, Nicholas A, Yukhananov R, Bu J, Wu CK, and Geula C. (2011). Age-related loss of calcium buffering and selective neuronal vulnerability in Alzheimer's disease. Acta. Neuropathol. 122, 565‒576.
Roberson ED, Scearce-Levie K, Palop JJ, Yan F, Cheng IH, Wu T, Gerstein H, Yu GQ, and Mucke L. (2007). Reducing endogenous tau ameliorates amyloid beta-induced deficits in an Alzheimer's disease mouse model. Science 316, 750-754.
Rockenstein E, Torrance M, Adame A, Mante M, Bar-on P, Rose JB, Crews L, and Masliah E. (2007). Neuroprotective effects of regulators of the glycogen synthase kinase-3beta signaling pathway in a transgenic model of Alzheimer's disease are associated with reduced amyloid precursor protein phosphorylation. J. Neurosci. 27, 1981-1991.
Ryder J, Su Y, Liu F, Li B, Zhou Y, and Ni B. (2003). Divergent roles of GSK3 and CDK5 in APP processing. Biochem Biophys Res Commun 312, 922-929.
Sahara N, Vega IE, Ishizawa T, Lewis J, McGowan E, Hutton M, Dickson D, and Yen SH. (2004). Phosphorylated p38MAPK specific antibodies cross-react with sarkosyl-insoluble hyperphosphorylated tau proteins. J. Neurochem. 90, 829‒838.
Saitoh M, Kunitomo J, Kimura E, Hayase Y, Kobayashi H, Uchiyama N, Kawamoto T, Tanaka T, Mol CD, Dougan DR, Textor GS, Snell GP, and Itoh F. (2009a). Design, synthesis and structure-activity relationships of 1,3,4-oxadiazole derivatives as novel inhibitors of glycogen synthase kinase-3beta. Bioorg Med Chem. 17, 2017-29.
Saitoh M, Kunitomo J, Kimura E, Iwashita H, Uno Y, Onishi T, Uchiyama N, Kawamoto T, Tanaka T, Mol CD, Dougan DR, Textor GP, Snell GP, Takizawa M, Itoh F, and Kori M.
(2009b) 2-{3-[4-(Alkylsulfinyl)phenyl]-1-benzofuran-5-yl}-5-methyl-1,3,4-oxadiazole derivatives as novel inhibitors of glycogen synthase kinase-3beta with good brain