Parkin expression in the adult mouse brain

• Published in: The European journal of neuroscience 2000-12, Vol.12 (12), p.4181-4194
• Main Authors: Stichel, C C, Augustin, M, Kühn, K, Zhu, X R, Engels, P, Ullmer, C, Lübbert, H
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Brain - cytology; Brain - metabolism; Cell Line; Cercopithecus aethiops; COS Cells; Humans; Ligases; Mice; Mice, Inbred C57BL; Molecular Sequence Data; Organ Specificity; Proteins - analysis; Proteins - chemistry; Proteins - genetics; Rats; Rats, Wistar; Recombinant Proteins - analysis; Recombinant Proteins - chemistry; Sequence Alignment; Sequence Homology, Amino Acid; Spinal Cord - cytology; Spinal Cord - metabolism; Transfection; Ubiquitin-Protein Ligases
• ISSN: 0953-816X; 1460-9568
• DOI: 10.1046/j.1460-9568.2000.01314.x

Mutations in a protein designated Parkin were shown to be involved in the pathogenesis of autosomal recessive juvenile parkinsonism. Nothing is known about its regional and subcellular distribution in the mouse. In order to elucidate the Parkin mRNA and protein distribution in the adult mouse, the mouse cDNA was cloned and polyclonal antisera were generated against the N-terminal part of mouse Parkin. The antibodies were shown to be specific using Western blot analysis, immunostaining of cells transfected with mouse Parkin and pre-absorption tests. The Parkin protein expression profile was studied using immunohistochemistry and Western blot analysis and was compared with that of the mRNA yielded by in situ hybridization and RT-PCR analysis. Parkin protein was widely distributed in all subdivisions of the mouse brain. Low levels were found in the telencephalon and diencephalon, while the brainstem contained a large number of cells heavily expressing Parkin. Ultrastructural analysis and double immunohistochemistry revealed that the majority of Parkin-expressing cells were neurons, while only single glial cells exhibited immunostaining. The protein was distributed nonhomogeneously throughout the entire cytoplasm. A subpopulation of Parkin-immunopositive cells displayed speckled immunodeposits in the nucleus. Dopaminergic cells of the substantia nigra pars compacta exhibited high levels of Parkin mRNA but no Parkin protein, while the striatum contained immunopositive profiles but no mRNA signals. Our data indicate that Parkin is neither restricted to a single functional system nor associated with a particular transmitter system. The speckled nuclear distribution of Parkin immunoreactivity strongly suggests a role for Parkin in gene expression.