Nothing Special   »   [go: up one dir, main page]

Academia.eduAcademia.edu

Memory and exploratory impairment in mice that lack the Park-2 gene and that over-express the human FTDP-17 mutant Tau

2008, Behavioural Brain Research

Available online at www.sciencedirect.com Behavioural Brain Research 189 (2008) 350–356 Research report Memory and exploratory impairment in mice that lack the Park-2 gene and that over-express the human FTDP-17 mutant Tau Paloma Navarro a , Rosa Guerrero a , Eva Gallego a , Jesus Avila b , Rosario Luquin c , Pedro J. Garcia Ruiz a , Marina P. Sanchez a,∗ a Fundacion Jimenez Diaz-Capio/Clinica Ntra. Sra. Concepcion, Universidad Autonoma de Madrid, 28040 Madrid, Spain b Centro de Biologia Molecular “Severo Ochoa”, CSIC/Universidad Autonoma de Madrid, Spain c Centro de Investigaciones Médicas Aplicadas, Universidad de Pamplona, Navarra, Spain Received 8 August 2007; received in revised form 17 January 2008; accepted 24 January 2008 Available online 5 February 2008 Abstract While mutations in the Park-2 gene are the most frequent cause of autosomal-recessive juvenile parkinsonism (AR-JP), they are also present in several forms of tauopathies. Conversely, in some forms of parkinsonism, mutations in the tau gene have also been observed. Deletion of the Park-2 gene and over-expression of mutant tau independently produce mild brain alterations in mice. However, the presence of both mutations simultaneously causes a tau neuropathology, involving reactive astrocytosis, neuron loss in the cortex and hippocampus, and lesions in nigrostriatal and motor neurons. Furthermore, mutant tau over-expression in mice produces important memory impairment. When “parkin” function was abolished in young tau transgenic mice, the memory alterations were exaggerated. Moreover, additional exploratory and motor deficits were observed in older mice, causing the memory alterations to be underestimated. Thus, while memory deficits are more severe in young mice they were somehow attenuated by exploratory impairments in ageing mutants. This double mutant animal will serve as a useful experimental tool to investigate the abnormal processing of hyperphosphorylated tau and its relationship to the development of the cognitive deficits associated with certain neurodegenerative diseases. © 2008 Elsevier B.V. All rights reserved. Keywords: Parkin; Park-2 gene deletion; Human tau transgene; FTDP tau mutations; Mouse model; Object recognition task; Memory alterations 1. Introduction The Park-2 gene encodes the ubiquitin protein ligase parkin that is implicated in proteasome-dependent protein degradation [47]. Loss-of-function mutations in the Park-2 gene cause autosomal-recessive juvenile parkinsonism (ARJP), one of the most common forms of familial parkinsonism [1,18,25,28,32,56,57] that is characterized by a selective loss of dopaminergic neurons in the substantia nigra without the appearance of Lewy bodies [19,35,56,57]. Single parkin mutations have been detected in patients with Parkinson’s disease (PD) [58], while mutations in the Park-2 gene have also been found ∗ Corresponding author at: Laboratory of Neurology, Fundacion Jimenez Diaz-Capio/Clinica Ntra. Sra. Concepción, Av Reyes Catolicos 2, 28040 Madrid, Spain. Tel.: +34 91 5504800x3251; fax: +34 91 5497700. E-mail address: msanchezg@fjd.es (M.P. Sanchez). 0166-4328/$ – see front matter © 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.bbr.2008.01.017 in patients with certain tauopathies [34,43]. Conversely, mutations in the gene encoding the microtubule-associated protein tau have been reported in other forms of familial parkinsonism and dementia [49]. Indeed, missense mutations of the tau gene cause autosomal-dominant frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) [20,41,50], and tau aggregates are also present in some other cases of autosomaldominant parkinsonism [59]. However, mutations in the tau gene have not been observed in the most frequent cases of tauopathies [52]. Moreover, several cases of dementia have been reported where mutations in Park-2 appear in conjunction with a tau protein pathology [34,35,56]. Thus, parkin may be functionally related to tau, and the combination of both parkin and tau mutations may trigger functional deficits, possibly related to the processing and degradation of anomalous hyperphosphorylated tau protein [26,40,43]. Moreover, abnormal parkin activity may contribute to the development of tauopathies in individuals with genetic risk factors [34,43]. P. Navarro et al. / Behavioural Brain Research 189 (2008) 350–356 Neurofibrillary tangles (NFTs) are neuronal inclusions that are composed of modified hyperphosphorylated tau protein, which has a low affinity for tubulin and a high tendency to selfassemble [3,6,10,11,16,22]. Clearance of the tau aggregates is mediated by the proteasome [55] and it is dependent on the phosphorylation state of tau [27]. In Alzheimer’s disease (AD), NFTs occur in the hippocampus, amygdala, the entorhinal and other association cortices, as well as in the basal forebrain. These brain areas are severely affected by the loss of neurons and synapses that contribute to the cognitive deficits and dementia in AD [5,12,46,59]. Transgenic mice and gene-targeting of Tau are valuable models to study these conditions, as they reproduce various aspects of tauopathies and AD tau pathology with associated cognitive changes [2,8,13–15,21,29,38,44,53,54]. Deletion of the parkin gene in Park− mice alters protein degradation and dopamine neurotransmission, without a loss of nigral dopaminergic neurons [24]. In FTDP-17 TauVLW mutant mice, the hTauVLW transgene is expressed in the cytoplasm and dendrites of cortical and hippocampal neurons, as well as in other brain regions not relevant to AD [30]. Furthermore, there is an age-dependent appearance of hyperphosphorylated tau filaments in mutant FTDP-17 tau-positive neurons, with the appearance of pretangles [30]. We generated a double mutant mouse (Park− /TauVLW ) that over-expresses a human FTDP-17 mutant four-repeat form of tau, the hTauVLW transgene, and that is also deficient in the Park-2 gene [17]. These mice develop normally, although their survival is considerably curtailed. The lack of the parkin protein in young TauVLW mice produces an accumulation of AT8 and PHF-1 hyperphosphorylated tau in cortical and hippocampal neurons. However, while the product of the hTauVLW transgene is found in both cell bodies and dendrites in young transgenic mice, it is only detected in the cell bodies of neurons when the Park-2 gene is deleted. Closer analysis of the Park− /TauVLW mouse brain indicates that either the subcellular distribution of the hTauVLW transgene product or its level of expression differs from that in the TauVLW mouse. Thus, in the absence of parkin, the mutant tau is excluded from the dendrites and it is restricted to the cytoplasm of neurons. Additionally, the presence of reactive astrocytes in these double mutants is associated with an increase in glial fibrillary acidic protein (GFAP) expression. Although very few apoptotic neurons were detected in single TauVLW mice, neuron loss is evident in the cortex and hippocampus of young Park− /TauVLW mice [17]. Moreover, in cortical and hippocampal extracts from double Park− /TauVLW mutants the expression of the neuroprotective heat shock protein 70 (Hsp70) was accentuated while the carboxyl-terminus of the Hsp70-interacting protein (CHIP) was depleted, a ubiquitin protein ligase known to attenuate the aggregation of tau [42] and to enhance the ubiquitination of phosphorylated tau [39,48]. When these animals died at the age of 14 months old, most neurons lacked tau and displayed a degenerated cytoplasm and condensed nuclei [17]. In addition, behavioural studies of Park− /TauVLW mice showed mild changes in motor activity (manuscript in preparation). Here, we have investigated the effect that the alterations in the brain of these Park− /TauVLW mice has on memory. Accordingly, we found that young animals over-expressing the 351 hTauVLW transgene develop severe memory deficits that were more pronounced when the Park-2 gene was deleted. Moreover, ageing double mutant mice present an additional defect in exploratory performance that could mask the expression of the more important memory impairments. These results further reflect the existence of a functional interaction between the parkin and tau proteins. 2. Methods 2.1. hTauVLW transgenic and parkin-null double mutant mice The human hTauVLW transgenic mouse line (TauVLW ) that over-expresses a four-repeat human tau isoform with the FTDP-17 mutations G272V, P301L and R406W has been described elsewhere [30]. The Parkin-deficient mice were generated by gene targeting of the Park-2 gene in embryonic stem cells [24]. Subsequently, we obtained Park− , TauVLW , Park− /TauVLW by breeding TauVLW mice with Park− or wild type mice on the same C57BL/6J genetic background [17]. All the experiments carried out on these mice were previously approved by the Animal Investigation Committee at our Institution. 2.2. Expression of mutant proteins in Western blots Protein extracts were prepared from brain homogenates in the presence of a protease inhibitor cocktail (Boehringer Manheim). Protein samples were separated on SDS-polyacrylamide gels, electroblotted onto nitrocellulose sheets, and incubated with specific antibodies that were detected by ECL (Amersham). The antibodies used to assess the expression of mutant proteins included: a parkin polyclonal antiserum directed against a peptide corresponding to residues 71–91 of mouse parkin [24]; T14 (Zymed Laboratories, INC), an antibody that recognises an epitope present in human tau protein but that is absent in endogenous murine tau; and the 7.51 antibody (a kind gift of Dr. C.M. Wischick, MRC, Cambridge, UK) that recognizes a phosphorylation-independent epitope at the carboxy-terminal region of human and murine tau molecules. 2.3. Object recognition task (ORT) We used the ORT to measure episodic memory retention in groups of young and adult mice. After a 10 min session of adaptation to an empty dark box, the time employed in exploring (tA and tB ) two equivalent objects (objects A and B) was measured for each mouse in the sample familiarisation phase. Mice whose total exploration time was ≤3 s were eliminated for the rest of the test. The test phase was then performed 2 h later, and the times (tA and tC ) the animal spent exploring two objects were recorded, the familiar object (A) and a novel object (C). Mice with a total exploration time ≤1 s per object in the test phase were also eliminated and the percentage time exploring objects A and C was then represented for the single and double mutant mice. A discrimination index (DI) was calculated as the ratio of the difference between the exploration times of the new (tC ) and the old familiar object (tA ), with respect to the total (tT ) exploration time as follows: DI = tC − tA /tT . 2.4. Statistical analysis Two groups of male animals were analyzed at 6–8 and 15–17 months of age, as representative samples of adult and aged mice. Initially, the number of animals in each group was 12–26. The values are expressed as means ± S.E.M. and the statistical evaluation was performed by one way-ANOVA followed by post hoc Newman–Keuls test (GraphPad Prism 2.0). Statistical significance was considered when p < 0.05. 3. Results and discussion Several cases of tauopathies have been reported in patients carrying mutations in the Park-2 gene while conversely, muta- 352 P. Navarro et al. / Behavioural Brain Research 189 (2008) 350–356 tions in the tau gene have been detected in patients with certain types of parkinsonism. Thus, parkin may be functionally related with tau, probably in the processing and degradation of hyperphosphorylated forms of the tau protein [26,39,43]. We described a patient who was carrier of a single mutation in the Park-2 gene, and who was homozygous for the H1/H1 haplotype in the tau gene, a risk haplotype for progressive supranuclear palsy [34,43]. Neuropathological studies on this patient identified tau aggregates and abnormal hyperphosphorylation of tau in these aggregates, suggesting that a partial deficit of parkin may trigger a tau pathology in individuals with molecular genetic risk factors [43]. This led us to postulate that parkin plays a role in the intracellular processing of tau and that abnormal parkin function may be a risk factor for the development of tauopathies. To test this possibility we generated a double mutant mouse in which the Park-2 gene was deleted [24] in conjunction with the presence of the human TauVLW transgene [30], the Park− /TauVLW mouse [17]. In the absence of parkin, there were mild nigrostriatal and motor neuron lesions in the hTauVLW transgenic mice [33]. Moreover, hyperphosphorylated tau aggregates with AT-8 (Fig. 1A) and PHF1 were detected [17], and GFAP immunostaining identified reactive astrocytes in the cortex and hippocampus (B). DNA fragmentation and a varying degree of cerebral atrophy were also observed in these regions [17]. Additionally, the expression of GFAP, Hsp70 and CHIP was altered in protein extracts from these animals [17]. The absence of parkin protein in single Park− and double Park− /TauVLW mutants was demonstrated in Western blots (C). Similarly, immunoblotting with the T14 antibody, which recognizes an epitope present in the human mutant tau, and with 7.51 that recognizes a phosphorylation-independent epitope at the carboxy-terminal region of human and murine tau molecules, revealed the existence of the two forms of tau in the brains of single TauVLW and double Park− /TauVLW mutant mice (C). We studied the functional effects of the Park-2 gene deletion in TauVLW transgenic mice in a memory performance test. Two groups of male animals were analyzed at 6–8 and 15–17 months of age and the effects of the mutations on the episodic memory of our mice were studied by measuring object recognition Fig. 1. Expression of AT8 phosphorylated tau, GFAP, parkin, and endogenous murine and human mutant tau in the cerebral cortex and hippocampus of Park− /TauVLW mice. AT8 immunoreactive tau aggregates are shown in some scattered neurons of the anterior frontal cortex of 14-month-old Park− /TauVLW mice (A). Reactive gliosis, identified with GFAP antibody, is shown in the dentate gyrus of double mutants at the same age (B). Representative Western blots of protein extracts from the cortex of wild type (1), Park− (2), TauVLW (3) and Park− /TauVLW (4) mice probed with the T14, 7.51 and parkin antibodies (C). Scale bar corresponds to 40 ␮m in (A) and to 60.4 ␮m in (B). P. Navarro et al. / Behavioural Brain Research 189 (2008) 350–356 learning in the ORT. The total exploration times of equivalent objects in the sample phase were measured in the three mutant mouse lines: Park− , TauVLW , Park− /TauVLW . At 6–8 months of age, the total exploration time in the sample phase for objects A and B was significantly higher in Park− mice than in wild type mice and those over-expressing the hTauVLW transgene (Fig. 2A). The percentage time that the three mutant groups of mice spent exploring objects A and B in this phase was similar to that of the wild type mice (B). By contrast, in the test phase the total exploration time of Park− mice of the familiar (A) and novel (C) objects was significantly higher than that of wild type mice and those over-expressing the hTauVLW transgene (C). Moreover, the Park− mice explored the novel object (C) longer than the familiar one (A) during the test phase, like the wild type animals (D). However, the over-expression of hTauVLW induced a failure to discriminate between the novel and the familiar objects (D), indicating the existence of deficits in the retention of episodic memory when the hTauVLW transgene is over-expressed. In accordance, the discrimination index decreased when hTauVLW was over-expressed and this alteration was slightly more pronounced when Park-2 gene was also deleted (E). 353 At 15–17 months of age, the total exploration time of TauVLW and Park− /TauVLW mice in the sample phase (objects A and B) was less than that of Park− and wild type mice (Fig. 3A). However, this difference was only statistically significant when compared to Park− mice, since deletion of the Park-2 gene produced a mild additional increase in the object exploration time (A). A small number of Park− /TauVLW animals explored the objects in the sample phase, although there was no difference in the percentage time they spent exploring objects A and B (B). A deficit was observed in object recognition memory in the TauVLW and Park− /TauVLW mice, reflected by the similar proportion of time they spent exploring objects A and C (D). Indeed, there was a decrease in the discrimination index in the three mutant groups when compared to the wild type animals (E), although this difference was only statistically significant for the TauVLW mice. In the aged Park− /TauVLW mice, the failure to detect a significant decrease was probably due to the limited number of animals that displayed exploratory behaviour during the test phase. Indeed, deletion of the Park-2 gene produced an increase in locomotor activity of both single and double Park− /TauVLW mice (manuscript in preparation). Interestingly, Park− mice showed a significant increase in object exploration time in the ORT, prob- Fig. 2. Analysis of episodic memory using the object recognition task (ORT) on young mice (6–8 months of age). Initially, 14, 12, 19 and 26 animals were analyzed from the wild type, Park− , TauVLW and Park− /TauVLW groups, respectively. (A) Total exploration time (tA + tB ) of two equal objects (objects A and B) in the sample phase, after the elimination of animals with a total exploration time ≤3 s. The numbers of mice finally analyzed in the sample phase were n = 9, 12, 15 and 23 for wild type, Park− , TauVLW and Park− /TauVLW mouse lines, respectively. (B) Percentage exploration of objects A and B in the sample phase. (C) Total exploration time (tA + tC ) of two different objects (A and C) in the test phase. The number of animals of the final groups in the test phase were n = 6, 11, 12 and 15 from wild type, Park− , TauVLW and Park− /TauVLW mouse lines, respectively. (D) Percentage time exploring two different objects (objects A and C) in the test phase after the elimination of animals with a total exploration time ≤1 s per object. (E) The DI was calculated as the ratio of the difference between the exploration times of the new (tC ) and the familiar object (tA ), and the total (tT ) exploration time (DI = tC − tA /tT ). Values are expressed as mean ± S.E.M. One way-ANOVA followed by Newman–Keuls tests were performed. *p < 0.05, **p < 0.01, ***p < 0.001. 354 P. Navarro et al. / Behavioural Brain Research 189 (2008) 350–356 Fig. 3. Analysis of episodic memory in mice aged (15–17 months of age) using the object recognition task. Initially, we analyzed 16, 15, 14 and 9 animals from the wild type, Park− , TauVLW and Park− /TauVLW groups, respectively. In the sample phase, after elimination of the animals with a total exploration time ≤3 s, 14, 14, 12 and 6 mice were analyzed from the wild type, Park− , TauVLW and Park− /TauVLW mouse lines, respectively. (A) Total exploration time (tT ) in the sample phase. (B) Percentage exploration times of objects A and B in the sample phase. (C) Total exploration time in the test phase. (D) Percentage time exploring two different objects (objects A and C) in the test phase after the elimination of animals with a total exploration time ≤1 s. The final number of animals used in the test phase were n = 13, 14, 9 and 4 from wild type (81.25%), Park− (93.3%), TauVLW (64.2%) and Park− /TauVLW (44%) mouse lines, respectively. (E) The DI is the ratio of the difference between the exploration times of the new (tC ) and the familiar object (tA ), and the total (tT ) exploration time (DI = tC − tA /tT ). Values are expressed as mean ± S.E.M. Statistical analysis was performed by one way-ANOVA followed by Newman–Keuls tests. *p < 0.05; **p < 0.01. ably as a consequence of their hyperactivity. However, while Park− /TauVLW mice moved around the cage and were active, they spent less time exploring the objects and most of them, the most affected population, had to be excluded from the test. It has been reported that a significant number of tau transgenic mouse models present neurofibrillary pathology, neuronal loss and memory deficits [2,4,13,15,23,38,44,45]. Moreover, improved animal models for neurodegenerative disorders have been generated by combining mutations of genes that interact in different pathways [7,9,14,29,31,36,37,51]. However, none of these mouse models generated to date fully recapitulate the neuropathological spectrum of tangle pathology observed in AD. The Park− /TauVLW mutant mice that we have produced will help us investigate the relationship between parkin and tau proteins in the development of tau pathology and the functional consequences of these alterations. The deletion of parkin alone did not produce any effect on memory performance in mice, while human mutant tau over-expression produced severe memory impairments. However, the combination of both parkin and tau mutations gives rise to a stronger memory impairment in young mice, although this effect is diminished in older mice, probably due to some exploratory and/or motor deficits that precede their premature death. The presence of tau aggregates in the cortex and hippocampus of young mice lacking parkin and those that over-express human mutant tau [17] could be the responsible for the increased memory impairment. The absence of parkin could interfere with the binding of tau to microtubules and as such, this may increase the availability of free cytosolic tau which is more susceptible to phosphorylation. Hence, parkin is probably implicated in the processing and degradation of hyperphosphorylated forms of the tau protein, although this process does not involve a direct interaction between the two proteins. Moreover, the existence of lesions in nigrostriatal and motor neurons at early ages [33] could be associated to the presence of motor disturbances that might interfere with the interpretation of the data obtained in the memory test. Further behavioural studies are now in progress to test this possibility. Nevertheless, the mouse model that we have generated is a useful tool to study the role played by the parkin ubiquitin ligase in the degradation of hyperphosphorylated tau, and in the ensuing formation of tau aggregates. Further studies of these mutants will hopefully help us to understand the mechanisms responsible for the memory impairments developed in some neurodegenerative diseases that are related to the tau and parkin proteins. P. Navarro et al. / Behavioural Brain Research 189 (2008) 350–356 Disclosure statement All the authors state that there are no actual or potential conflicts of interest including any financial, personal or other relationships with individuals or organizations that could inappropriately influence our work. Acknowledgments The authors would like to thank Drs. Jeremy Park, Jesus Benavides, and Filip Lim for the use of Park− and TauVLW mice, and Drs. Francisco Wandosell and Javier Dı́az-Nido for critical reading of the manuscript. We are also grateful to the Animal Facility of the Fundación Jiménez Dı́az-Capio, especially to M. Luisa Valbuena, for technical support. This work was supported by a grant from the Fondo de Investigaciones Sanitarias. MPS was supported by the Instituto de Salud Carlos III and FJD-Capio, and PN was supported by a fellowship from the Fundación Conchita Rábago. References [1] Abbas N, Lucking CB, Ricard S, Durr A, Bonifati V, De Michele G, et al. A wide variety of mutations in the parkin gene are responsible for autosomal recessive parkinsonism in Europe. French Parkinson’s Disease Genetics Study Group and the European Consortium on Genetic Susceptibility in Parkinson’s Disease. Hum Mol Genet 1999;8:567–74. [2] Allen B, Ingram E, Takao M, Smith MJ, Jakes R, Virdee K, et al. Abundant tau filaments and nonapoptotic neurodegeneration in transgenic mice expressing human P301S tau protein. J Neurosci 2002;22:9340–51. [3] Alonso A, Zaidi T, Novak M, Grundke-Iqbal I, Iqbal K. Hyperphosphorylation induces self-assembly of tau into tangles of paired helical filaments/straight filaments. Proc Natl Acad Sci USA 2001;98:6923–8. [4] Arendash GW, Lewis J, Leighty RE, McGowan E, Cracchiolo J-R, Hutton M, et al. Multi-metric behavioral comparison of APPsw and P301L models for Alzheimer’s disease: linkage of poorer cognitive performance to tau pathology in forebrain. Brain Res 2004;1012:29–41. [5] Arendt T, Bigl V, Arendt A, Tennstedt A. Loss of neurons in the nucleus basalis of Meynert in Alzheimer’s disease, paralysis agitans and Korsakoff’s Disease. Acta Neuropathol (Berl) 1983;61:101–8. [6] Bramblett GT, Goeder M, Jakes R, Merrick SE, Trojanowski JQ, Lee VM. Abnormal tau phosphorylation at Ser396 in Alzheimer’s disease recapitulates development and contributes to reduced microtubule binding. Neuron 1993;10:1089–99. [7] Brecht WJ, Harris FM, Chang S, Tesseur I, Yu GQ, Xu Q, et al. Neuronspecific apolipoprotein e4 proteolysis is associated with increased tau phosphorylation in brains of transgenic mice. J Neurosci 2004;24:2527– 34. [8] Brion JP, Tremp G, Octave JN. Transgenic expression of the shortest human tau affects its compartmentalization and its phosphorylation as in the pretangle stage of Alzheimer’s disease. Am J Pathol 1999;154:255–70. [9] Duff K, Knight H, Refolo LM, Sanders S, Yu X, Picciano M, et al. Characterization of pathology in transgenic mice over-expressing human genomic and cDNA tau transgenes. Neurobiol Dis 2000;7:87–98. [10] Feany MB, Dickson DW. Neurodegenerative disorders with extensive tau pathology: a comparative study and review. Ann Neurol 1996;40:139–48. [11] Goedert M, Spillantini MG, Cairns NJ, Crowther RA. Tau proteins of Alzheimer paired helical filaments: abnormal phosphorylation of all six brain isoforms. Neuron 1992;8:159–68. [12] Gomez-Isla T, Price JL, McKeel Jr DW, Morris JC, Growdon JH, Hyman BT. Profound loss of layer II entorhinal cortex neurons occurs in very mild Alzheimer’s disease. J Neurosci 1996;16:4491–500. [13] Gotz J, Chen F, Barmettler R, Nitsch RM. Tau filament formation in transgenic mice expressing P301L tau. J Biol Chem 2001;276:529–34. 355 [14] Gotz J, Chen F, van Dorpe J, Nitsch RM. Formation of neurofibrillary tangles in P301L tau transgenic mice induced by A␤ 42 fibrils. Science 2001;293:1491–5. [15] Gotz J, Tolnay M, Barmettler R, Chen F, Probst A, Nitsch RM. Oligodendroglial tau filament formation in transgenic mice expressing G272V tau. Eur J Neurosci 2001;13:2131–40. [16] Grundke-Iqbal I, Iqbal K, Tung YC, Quinlan M, Wisniewski HM, Binder LI. Abnormal phosphorylation of the microtubule-associated protein tau in Alzheimer cytoskeletal pathology. Proc Natl Acad Sci USA 1986;83:4913–7. [17] Guerrero R, Navarro P, Gallego E, Avila J, Garcı́a de Yébenes J, Sanchez, MP. Park2-null/tau transgenic mice reveal an interaction between parkin and tau. J Alzheimers Dis; March 2008. [18] Hattori N, Kitada T, Matsumine H, Asakawa S, Yamamura Y, Yoshino H, et al. Molecular genetic analysis of a novel Parkin gene in Japanese families with autosomal recessive juvenile parkinsonism: evidence for variable homozygous deletions in the Parkin gene in affected individuals. Ann Neurol 1998;44:935–41. [19] Hayashi S, Wakabayashi K, Ishikawa A, Nagai H, Saito M, Maruyama M, et al. An autopsy case of autosomal-recessive juvenile parkinsonism with a homozygous exon 4 deletion in the parkin gene. Mov Dis 2000;15:884–8. [20] Hutton M, Lendon CL, Rizzu P. Association of missense and 5 -splicesite mutations in tau with the inherited dementia FTDP-17. Nature 1998;393:702–5. [21] Ikeda M, Shoji M, Kawarai T, Kawarabayashi T, Matsubara E, Murakami T, et al. Accumulation of filamentous tau in the cerebral cortex of human tau R406W transgenic mice. Am J Pathol 2005;166:521–31. [22] Iqbal K, Grundke-Iqbal I. Discoveries of Tau, abnormally hyperphosphorylated tau and others of neurofibrillary degeneration: a personal historical perspective. J Alzheimers Dis 2006;9:219–42. [23] Ishihara T, Hong M, Zhang B, Nakagawa Y, Lee MK, Trojanowski JQ, Lee VM. Age-dependent emergence and progression of a tauopathy in transgenic mice overexpressing the shortest human tau isoform. Neuron 1999;24:751–62. [24] Itier JM, Ibanez P, Mena MA, Abbas N, Cohen-Salmon C, Bohme A, et al. Parkin gene inactivation alters behavior and dopamine neurotransmission in the mouse. Hum Mol Genet 2003;12:2277–91. [25] Kitada T, Asakawa S, Hattori N, Matsumine H, Yamamuram Y, Minoshima S, et al. Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism. Nature 1998;392:605–8. [26] Klein RL, Dayton RD, Henderson KM, Petrucelli L. Parkin is protective for substantia nigra dopamine neurons in a tau gene transfer neurodegeneration model. Neurosci Lett 2006;401:130–5. [27] Layfield R, Cavey JR, Lowe J. Role of ubiquitin-mediated proteolysis in the pathogenesis of neurodegenerative disorders. Ageing Res Rev 2003;2:343–56. [28] Leroy E, Anastasopoulos D, Konitsiotis S, Laveda C, Polymeropoulos MH. Deletions in the Parkin gene and genetic heterogeneity in a Greek family with early onset Parkinson’s disease. Hum Genet 1998;103:424–7. [29] Lewis J, Dickson DW, Lin WL, Chisholm L, Corral A, Jones G, et al. Enhanced neurofibrillary degeneration in transgenic mice expressing mutant tau and APP. Science 2001;293:1487–91. [30] Lim F, Lucas JJ, Gomez-Ramos P, Moran MA, Avila J. FTDP-17 mutations in tau transgenic mice provoke lysosomal abnormalities and Tau filaments in forebrain. Mol Cell Neurosci 2001;8:702–14. [31] Lucas JJ, Hernandez F, Gomez-Ramos P, Moran MA, Hen R, Avila J. Decreased nuclear ␤-catenin, tau hyperphosphorylation and neurodegeneration in GSK-3␤ conditional transgenic mice. EMBO J 2001;20:27–39. [32] Matsumine H, Saito M, Shimoda-Matsubayashi S. Localization of a gene for an autosomal recessive form of juvenile parkinsonism to chromosome 6q25.2-27. Am J Hum Genet 1997;60:588–96. [33] Menendez J, Rodriguez-Navarro JA, Solano RM, Casarejos MJ, Rodal I, Guerrero R, et al. Suppression of Parkin enhances nigrostriatal and motor neuron lesion in mice over-expressing human-mutated tau protein. Hum Mol Genet 2006;15:2045–58. [34] Morales B, Martinez A, Gonzalo I, Vidal L, Gomez-Tortosa E, Rabano A, et al. Steele–Richardson–Olszewski syndrome in a patient with a single C212Y mutation in the parkin protein. Mov Dis 2002;17:1374–80. 356 P. Navarro et al. / Behavioural Brain Research 189 (2008) 350–356 [35] Mori H, Kondo T, Yokochi M. Pathologic and biochemical studies of juvenile parkinsonism linked to chromosome 6q. Neurology 1998;51:890–2. [36] Oddo S, Caccamo A, Kitazawa M, Tseng BP, LaFerla FM. Amyloid deposition precedes tangle formation in a triple transgenic model of Alzheimer’s disease. Neurobiol Aging 2003;24:1063–70. [37] Oddo S, Caccamo A, Shepherd JD, Murphy MP, Golde TE, Kayed R, et al. Triple-transgenic model of Alzheimer’s disease with plaques and tangles: intracellular A␤ and synaptic dysfunction. Neuron 2003;39:409–21. [38] Pennanen L, Wolfer DP, Nitsch RM, Götz J. Impaired spatial reference memory and increased exploratory behaviour in P301L transgenic mice. Genes Brain Behav 2006;5:369–79. [39] Petrucelli L, Dickson D, Kehoe K, Taylor J, Snyder H, Grover A, et al. CHIP and Hsp70 regulate tau ubiquitination, degradation and aggregation. Hum Mol Genet 2004;13:703–14. [40] Petrucelli LK, Lockhart P. Tau is a parkin substrate; implications for neurodegenerative disease. Mov Dis 2002;17:S156. [41] Poorkaj P, Bird TD, Wijsman E. Tau is a candidate gene for chromosome 17 frontotemporal dementia. Ann Neurol 1998;43:815–25. [42] Sahara N, Murayama M, Mizoroki T, Urushitani M, Imai Y, Takahashi R, et al. In vivo evidence of CHIP up-regulation attenuating tau aggregation. J Neurochem 2005;94:1254–63. [43] Sanchez MP, Gonzalo I, Avila J, Garcı́a de Yébenes J. Progressive supranuclear palsy and tau hyperphosphorylation in a patient with a C212Y parkin mutation. J Alzheimers Dis 2002;4:399–404. [44] SantaCruz KS, Paulson J, Lewis J, Mariash A, Forster C, Guimaraes A, et al. Cell loss and gliosis in a transgenic mouse with regulatable P301L tau overexpression. Poster 840.25 presented at the society for neuroscience meeting, New Orleans, AT. [45] Schindowski K, Bretteville A, Leroy K, Bégard S, Brion JP, Hamdane M, Buée L. Alzheimer’s disease-like tau neuropathology leads to memory deficits and loss of functional synapses in a novel mutated tau transgenic mouse without any motor deficits. Am J Pathol 2006;169:599–616. [46] Scott SA, DeKosky ST, Sparks DL, Knox CA, Scheff SW. Amygdala cell loss and atrophy in Alzheimer’s disease. Ann Neurol 1992;32:555–63. [47] Shimura H, Hattori N, Kubo S. Familial Parkinson disease gene product, parkin, is an ubiquitin-protein ligase. Nat Genet 2000;25:302–5. [48] Shimura H, Schwartz D, Gygi SP, Kosik KS. CHIP–Hsc70 complex ubiquitinates phosphorylated tau and enhances cell survival. J Biol Chem 2004;279:4869–76. [49] Spillantini MG. Tau and Parkinson disease. J Am Med Assoc 2001;286:2324–6. [50] Spillantini MG, Bird TD, Ghetti B. Frontotemporal dementia and Parkinsonism linked to chromosome 17: a new group of tauopathies. Brain Pathol 1998;8:387–402. [51] Spires TL, Hyman BT. Transgenic models of Alzheimer’s disease: learning from animals. Neuro Rx 2005;2:423–37. [52] Stanford PM, Brooks WS, Teber ET. Frequency of tau mutations in familial and sporadic frontotemporal dementia and other tauopathies. J Neurol 2004;251:1098–104. [53] Tanemura K, Murayama M, Akagi T, Hashikawa T, Tominaga T, Ichikawa M, et al. Neurodegeneration with tau accumulation in a transgenic mouse expressing V337M human tau. J Neurosci 2002;22:133–41. [54] Tatebayashi Y, Miyasaka T, Chui DH, Akagi T, Mishima K, Iwasaki K, et al. Tau filament formation and associative memory deficit in aged mice expressing mutant (R406W) human tau. Proc Natl Acad Sci USA 2002;99:13896–901. [55] van Leeuwen FW, Hol EM, Fischer DF. Frameshift proteins in Alzheimer’s disease and in other conformational disorders: time for the ubiquitin–proteasome system. J Alzheimers Dis 2006;9:319–25. [56] van De Warrenburg BP, Lammens M, Lucking CB. Clinical and pathologic abnormalities in a family with parkinsonism and parkin gene mutations. Neurology 2001;56:555–7. [57] von Coelln R, Dawson VL, Dawson TM. Parkin-associated Parkinson’s disease. Cell Tissue Res 2004;318:175–84. [58] West A, Lincoln S, Lucking CB, Nicholl D, Bonifati V, Rawal N, et al. French Parkinson’s Disease Genetics Study Group and the European Consortium on Genetic Susceptibility on Parkinson’s Disease. Complex relationship between Parkin mutations and Parkinson disease. Am J Med Genet 2002;114:584–91. [59] Wszolek ZK, Pfeiffer RF, Tsuboi Y, Uitti RJ, McComb RD, Stoessl AJ, et al. Autosomal dominant parkinsonism associated with variable synuclein and tau pathology. Neurology 2004;62:1619–22.