Formation of Neurofibrillary Tangles in P301L Tau Transgenic Mice Induced by Aβ42 Fibrils
Author:
Affiliation:
1. Division of Psychiatry Research, University of Zürich, August Forel Strasse 1, 8008 Zürich, Switzerland.
2. Experimental Genetics Group, Center for Human Genetics, K. U. Leuven, Campus Gasthuisberg, Leuven, Belgium.
Abstract
Publisher
American Association for the Advancement of Science (AAAS)
Subject
Multidisciplinary
Reference38 articles.
1. Neurofibrillary tangles, amyotrophy and progressive motor disturbance in mice expressing mutant (P301L) tau protein
2. Tau Filament Formation in Transgenic Mice Expressing P301L Tau
3. Tau and transgenic animal models
4. To generate transgenic mice we introduced the human pathogenic tau mutation P301L into the cDNA encoding the longest human brain tau isoform by a polymerase chain reaction (PCR)–mediated approach. This isoform contained exons 2 and 3 as well as four microtubule-binding repeats (2 + 3 + 4R htau40). To discriminate P301L tau transgenic from wild-type tau transgenic mice we introduced a silent mutation into the P301L construct that destroys a diagnostic Sma I restriction site. The cDNA was conferred with a Kozak consensus sequence and was subcloned into a murine Thy.1.2 genomic expression vector (provided by H. van der Putten Novartis Basel). Vector sequences of this construct (named pR5) were removed before microinjection. Transgenic mice were produced by pronuclear injection of B6D2F1 × B6D2F1 embryos. Founders were identified by PCR analysis of lysates from tail biopsies with two different primer pairs. Founder animals were intercrossed with C57BL/6 mice to establish lines. Transgenic mice were screened with oligonucleotides tau-I (5′-GGAGTTCGAAGTGATGGAAG-3′) and tau-K (5′-GGTTTTTGCTGGAATCCTGG-3′) and yielded an amplification product of 500 base pairs. A restriction digest of the amplification product by Sma I confirmed the presence of the P301L transgene. Of 10 independent transgenic lines 4 had comparable expression levels as determined by immunoblot analysis. Line pR5-183 was used in the present study.
5. K. B. J. F. a. G. Paxinos The Mouse Brain in Stereotaxic Coordinates (Academic Press San Diego 1997).
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