Specht, Christian G.; (2002) Characterisation of an inbred mouse strain with a deletion of the [alpha]-synuclein locus. Doctoral thesis (Ph.D.), University College London.

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Abstract

mRNA expression profiling was performed on a knock-in mouse model with a mutation in the NMDA receptor subunit NRl (N598R) that affects the receptors' function as coincidence detector. This approach was aimed at identifying downstream effects produced by the Ca2+ influx through NMDA receptors at the level of gene expression. cDNA array technology revealed striking differences only in the mRNA expression level of [alpha]-synuclein, a protein that has been implicated in the pathophysiology of a range of neurodegenerative diseases. However, this was not caused by the NRl mutation, but by a chromosomal deletion of the [alpha]-synuclein gene locus in the C57BL/6J inbred mice that were used for backcrossing the mutant strain. The deletion was shown to be present only in a subpopulation of C57BL/6J mice, now- referred to as C57BL/6JOlaHsd-Del(6)Snca1Slab. In addition to [alpha]-synuclein, other genes may be affected by the deletion that is estimated to be 120-500 kb in size. [alpha]-synuclein-deficient animals appear phenotypically normal. They show no compensatory upregulation of other members of the synuclein family, namely [alpha]-synuclein and [alpha]-synuclein. Similarly, the expression of synphilin-1, a known interacting partner of [alpha]-synuclein was unaffected. The C57BL/6JOlaHsd-Del(6)SncalSlab mouse model should help in the understanding of the physiological function of [alpha]-synuclein and its involvement in synucleinopathies. Also, the findings exemplify unexpected complications that may arise during the study of transgenic models or inbred strains. A Sindbis virus system was developed for the expression of fluorescent [alpha]-synuclein fusion proteins in neurons. A range of recombinant virion preparations was tested in plaque assays and the expression of the recombinant proteins was characterised. Initial analysis of the expression of [alpha]-synuclein-eGFP in organotypic hippocampal neurons suggested that the protein accumulated in presynaptic locations. This approach could be used for the study of the subcellular localisation and of protein interactions of [alpha]-synuclein.

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