Mobility and Subcellular Localization of Endogenous, Gene-edited Tau Differs from That of Over-expressed Human Wild-type and P301L Mutant Tau

Overview

Journal Sci Rep

Specialty Science

Date 2016 Jul 6

PMID 27378256

Citations 21

Authors

Di Xia

Julia M Gutmann

Jurgen Gotz

Affiliations

Soon will be listed here.

Abstract

Alzheimer's disease (AD) and a subset of frontotemporal dementia termed FTLD-Tau are characterized by a massive, yet incompletely characterized and understood redistribution of Tau. To establish a framework for understanding this pathology, we used the genome-editing tool TALEN and generated Tau-mEOS2 knock-in mice to determine the mobility and subcellular localization of endogenous Tau in hippocampal cultures. We analysed Tau in axons, dendrites and spines at three stages of maturation using live-cell imaging, photo-conversion and FRAP assays. Tau-mEOS2 cultures were compared with those over-expressing EGFP-tagged forms of human wild-type (hWT-Tau) and P301L mutant Tau (hP301L-Tau), modelling Tau accumulation in AD and FTLD-Tau, respectively. In developing neurons, Tau-mEOS2 followed a proximo-distal gradient in axons and a subcellular distribution similar to that of endogenous Tau in neurons obtained from wild-type mice, which were abolished, when either hWT-Tau or hP301L-Tau was over-expressed. For the three conditions, FRAP analysis revealed a similar mobility in dendrites compared with axons; however, Tau-mEOS2 was less mobile than hWT-Tau and hP301L-Tau and the mobile fraction was smaller, possibly reflecting less efficient microtubule binding of Tau when over-expressed. Together, our study presents Tau-mEOS2 mice as a novel tool for the study of Tau in a physiological and a pathological context.

Citing Articles

Tau forms synaptic nano-biomolecular condensates controlling the dynamic clustering of recycling synaptic vesicles.

Longfield S, Mollazade M, Wallis T, Gormal R, Joensuu M, Wark J Nat Commun. 2023; 14(1):7277.

PMID: 37949856 PMC: 10638352. DOI: 10.1038/s41467-023-43130-4.

Clinical relevance of animal models in aging-related dementia research.

Padmanabhan P, Gotz J Nat Aging. 2023; 3(5):481-493.

PMID: 37202516 DOI: 10.1038/s43587-023-00402-4.

Dendritic spine loss deep in the neocortex and dendrite distortion with diffusion disturbances occur early in experimental pneumococcal meningitis.

Baronti D, Tomov N, Hupp S, Mitchell T, Iliev A Front Neurosci. 2023; 16:912445.

PMID: 36704002 PMC: 9871924. DOI: 10.3389/fnins.2022.912445.

Tau differentially regulates the transport of early endosomes and lysosomes.

Balabanian L, Lessard D, Swaminathan K, Yaninska P, Sebastien M, Wang S Mol Biol Cell. 2022; 33(13):ar128.

PMID: 36129768 PMC: 9634973. DOI: 10.1091/mbc.E22-01-0018.

Brain Cell Type-Specific Nuclear Proteomics Is Imperative to Resolve Neurodegenerative Disease Mechanisms.

Nelson R, Dammer E, Santiago J, Seyfried N, Rangaraju S Front Neurosci. 2022; 16:902146.

PMID: 35784845 PMC: 9243337. DOI: 10.3389/fnins.2022.902146.

References

Manley S, Gillette J, Patterson G, Shroff H, Hess H, Betzig E . High-density mapping of single-molecule trajectories with photoactivated localization microscopy. Nat Methods. 2008; 5(2):155-7. DOI: 10.1038/nmeth.1176. View

Querfurth H, LaFerla F . Alzheimer's disease. N Engl J Med. 2010; 362(4):329-44. DOI: 10.1056/NEJMra0909142. View

Sultan A, Nesslany F, Violet M, Begard S, Loyens A, Talahari S . Nuclear tau, a key player in neuronal DNA protection. J Biol Chem. 2010; 286(6):4566-75. PMC: 3039398. DOI: 10.1074/jbc.M110.199976. View

Goedert M, Spillantini M, Jakes R, Rutherford D, Crowther R . Multiple isoforms of human microtubule-associated protein tau: sequences and localization in neurofibrillary tangles of Alzheimer's disease. Neuron. 1989; 3(4):519-26. DOI: 10.1016/0896-6273(89)90210-9. View

Hinrichs M, Jalal A, Brenner B, Mandelkow E, Kumar S, Scholz T . Tau protein diffuses along the microtubule lattice. J Biol Chem. 2012; 287(46):38559-68. PMC: 3493901. DOI: 10.1074/jbc.M112.369785. View

Chew Y, Fan X, Gotz J, Nicholas H . Regulation of age-related structural integrity in neurons by protein with tau-like repeats (PTL-1) is cell autonomous. Sci Rep. 2014; 4:5185. PMC: 4046136. DOI: 10.1038/srep05185. View

Goedert M, Spillantini M, Potier M, Ulrich J, Crowther R . Cloning and sequencing of the cDNA encoding an isoform of microtubule-associated protein tau containing four tandem repeats: differential expression of tau protein mRNAs in human brain. EMBO J. 1989; 8(2):393-9. PMC: 400819. DOI: 10.1002/j.1460-2075.1989.tb03390.x. View

Neve R, Harris P, Kosik K, Kurnit D, Donlon T . Identification of cDNA clones for the human microtubule-associated protein tau and chromosomal localization of the genes for tau and microtubule-associated protein 2. Brain Res. 1986; 387(3):271-80. DOI: 10.1016/0169-328x(86)90033-1. View

Xia D, Li C, Gotz J . Pseudophosphorylation of Tau at distinct epitopes or the presence of the P301L mutation targets the microtubule-associated protein Tau to dendritic spines. Biochim Biophys Acta. 2015; 1852(5):913-24. DOI: 10.1016/j.bbadis.2014.12.017. View

10.

Dawson H, Ferreira A, Eyster M, Ghoshal N, Binder L, Vitek M . Inhibition of neuronal maturation in primary hippocampal neurons from tau deficient mice. J Cell Sci. 2001; 114(Pt 6):1179-87. DOI: 10.1242/jcs.114.6.1179. View

11.

Brion J, Smith C, Couck A, Gallo J, Anderton B . Developmental changes in tau phosphorylation: fetal tau is transiently phosphorylated in a manner similar to paired helical filament-tau characteristic of Alzheimer's disease. J Neurochem. 1993; 61(6):2071-80. DOI: 10.1111/j.1471-4159.1993.tb07444.x. View

12.

Chew Y, Fan X, Gotz J, Nicholas H . PTL-1 regulates neuronal integrity and lifespan in C. elegans. J Cell Sci. 2013; 126(Pt 9):2079-91. PMC: 4068611. DOI: 10.1242/jcs.jcs124404. View

13.

Wefers B, Panda S, Ortiz O, Brandl C, Hensler S, Hansen J . Generation of targeted mouse mutants by embryo microinjection of TALEN mRNA. Nat Protoc. 2013; 8(12):2355-79. DOI: 10.1038/nprot.2013.142. View

14.

Fath T, Ke Y, Gunning P, Gotz J, Ittner L . Primary support cultures of hippocampal and substantia nigra neurons. Nat Protoc. 2009; 4(1):78-85. DOI: 10.1038/nprot.2008.199. View

15.

Spillantini M, Crowther R, Kamphorst W, Heutink P, van Swieten J . Tau pathology in two Dutch families with mutations in the microtubule-binding region of tau. Am J Pathol. 1998; 153(5):1359-63. PMC: 1853390. DOI: 10.1016/S0002-9440(10)65721-5. View

16.

Lu J, Li T, He R, Bartlett P, Gotz J . Visualizing the microtubule-associated protein tau in the nucleus. Sci China Life Sci. 2014; 57(4):422-31. DOI: 10.1007/s11427-014-4635-0. View

17.

Kampers T, Pangalos M, Geerts H, Wiech H, Mandelkow E . Assembly of paired helical filaments from mouse tau: implications for the neurofibrillary pathology in transgenic mouse models for Alzheimer's disease. FEBS Lett. 1999; 451(1):39-44. DOI: 10.1016/s0014-5793(99)00522-0. View

18.

Yu D, Feinstein S, Valentine M . Effects of wild type tau and disease-linked tau mutations on microtubule organization and intracellular trafficking. J Biomech. 2015; 49(8):1280-1285. DOI: 10.1016/j.jbiomech.2015.09.043. View

19.

Janning D, Igaev M, Sundermann F, Bruhmann J, Beutel O, Heinisch J . Single-molecule tracking of tau reveals fast kiss-and-hop interaction with microtubules in living neurons. Mol Biol Cell. 2014; 25(22):3541-51. PMC: 4230615. DOI: 10.1091/mbc.E14-06-1099. View

20.

Rizzu P, van Swieten J, Joosse M, Hasegawa M, Stevens M, Tibben A . High prevalence of mutations in the microtubule-associated protein tau in a population study of frontotemporal dementia in the Netherlands. Am J Hum Genet. 1999; 64(2):414-21. PMC: 1377751. DOI: 10.1086/302256. View