Differential expression of the parkin gene in the human brain and peripheral leukocytes

doi:10.1016/S0304-3940(98)00697-1

Neuroscience Letters

Volume 254, Issue 3, 2 October 1998, Pages 180-182

https://doi.org/10.1016/S0304-3940(98)00697-1 Get rights and content

Abstract

Molecular cloning of the responsible gene on chromosome 6q25.2-27 for autosomal recessive juvenile parkinsonism (AR-JP) identified a novel protein of unknown function, named parkin. In patients with AR-JP, deletions most commonly involve exons 3–5 in the parkin gene. For mutation screening we tried to analyze the parkin transcript amplified by RT-PCR. Based on the assumption that illegitimate transcription of the parkin gene may occur in every cell type, we successfully amplified the parkin message from human peripheral leukocytes using RT-PCR. The parkin transcript in leukocytes was smaller in size than the full-length transcript in the brain. DNA sequencing determined that exons 3–5 were spliced out in the normal human leukocyte transcript. Our results demonstrate that alternative splicing produces distinct parkin transcripts in different tissues. Moreover, physiological splicing of deletion-prone exons may provide an important clue to understanding the pathogenesis of AR-JP.

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Acknowledgements

We thank Yukiko Takura for her expert technical assistance. This work was supported by the grants from the Sankyo Foundation of Life Science, the Science Research Promotion Fund from Japan Private School Promotion Foundation, Research Grant (8A-1and 8A-2) for Nervous and Mental Disorders from the Ministry of Health and Welfare, and Research Grants (08457195, 09470156, 09770460, 09877121 and10044319) from the Ministry of Education, Science, Sports and Culture.

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Parkin-dependent and -independent degradation of synaptotagmin-11 in neurons and astrocytes
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Citation Excerpt :
Ubiquitin-proteasome pathway (UPP) is critical for the regulation of short-lived proteins and clearance of misfolded and aggregated proteins, perturbation of which links to brain disorders including Parkinson’s disease (PD) [1,2]. Parkin, encoded by PARK2 gene (PD-linked genetic loci 2) [3], is an E3 ubiquitin-ligase enzyme in the UPP and is expressed in the central nervous system [4]. It plays multiple functions in mitochondrial homeostasis, autophagy and proteasomal degradation [5].
Synaptotagmin-11 (Syt11) is associated with schizophrenia and Parkinson’s disease (PD) and is a critical substrate of parkin, an E3 ubiquitin ligase linked to PD. Previously we reported that Syt11 regulates multiple membrane trafficking pathways in neurons and glia. However, the regulation of Syt11 degradation remains largely unknown. As the ubiquitin-proteasome pathway (UPP) plays crucial roles in protein degradation and quality control, we investigated UPP-dependent Syt11 degradation in this study. We found that Syt11 is a short-lived protein with a half-life of 1.49 h in the presence of a protein synthesis inhibitor cycloheximide and is mainly degraded by UPP in neurons. The degradation was further accelerated under sustained neuronal activity and was parkin-dependent. Interestingly, Syt11 had a faster turnover in astrocytes with a half-life of 0.58 h, and UPP partially contributed to its degradation. Mechanical stress applied on astrocytes by hypoosmotic treatment led to reduced Syt11 protein level but increased parkin level. However, the degradation of Syt11 was parkin-independent under both isoosmotic and hypoosmotic condition. Altogether, our results revealed active and distinct proteolytic regulation of Syt11 in neurons and astrocytes.
Alternative splicing of mRNA in the molecular pathology of neurodegenerative diseases
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Citation Excerpt :
There is limited information available in the literature about PARK2 splice variants in the human brain. The known PARK2 AS events include (i) an exon 4 splice variant (E4SV) found in both the substantia nigra (SN) and leukocytes; (ii) an exon 3–4–5 splice variant (E345SV) found in human leukocytes; and (iii) an exon 5 splice variant (E5SV) found in the brain (Kitada et al., 1998; Sunada et al., 1998; Tan et al., 2005). Mutations in the splice acceptor or splice donor sites of PARK2 introns 1, 2, 4, 5, 7, 8, 9, and 11 have been identified in the peripheral blood samples of PD sufferers (Bardien et al., 2009; Bertoli-Avella et al., 2005; Illarioshkin et al., 2003; Nuytemans et al., 2009; Pankratz et al., 2009; Pigullo et al., 2004; Scherfler et al., 2004).
Alternative splicing (AS) is a post-transcriptional process that occurs in multiexon genes, and errors in this process have been implicated in many human diseases. Until recently, technological limitations prevented AS from being examined at the genome-wide scale. With the advent of new technologies, including exon arrays and next-generation sequencing (NGS) techniques (e.g., RNA-Seq), a higher resolution view of the human transcriptome is now available. This is particularly applicable in the study of neurodegenerative brain diseases (NBDs), such as Alzheimer's disease and Parkinson's disease, because the brain has the greatest amount of alternative splicing of all human tissues. Although many of the AS events associated with these disorders were initially identified using low-throughput methodologies, genome-wide analysis allows for more in-depth studies, marking a new chapter in transcript exploration. In this review, the latest technologies used to study the transcriptome and the AS genes that have been associated with a number of neurodegenerative brain diseases are discussed.
Heterozygous exon 3 deletion in the Parkin gene in a patient with clinical and radiological MSA-C phenotype
2011, Clinical Neurology and Neurosurgery
Citation Excerpt :
A Parkin 40 bp heterozygous deletion in exon 3 segregating in an autosomal dominant fashion has been described in a parkinsonian family with Lewy body pathology, with a marked variation in clinical presentation, which illustrates how complex is the pathophysiology of those diseases associated with mutations in putative recessive genes [11]. Reviews of data on heterozygous Parkin mutations suggest that haploinsufficiency may be enough to cause the disease in a few cases [12], on the other hand, alternative splicing of Parkin exons 3–5 in peripheral leucocytes of healthy subjects has been reported [13]. We can speculate that alternative spliced isoforms will be deleterious only when expressed in the brain.
Expression analysis of suppression of tumorigenicity 13 gene in patients with Parkinson's disease
2010, Neuroscience Letters
Parkinson's disease (PD) is the second most common progressive neurodegenerative disorder characterized by the degeneration of dopaminergic neurons. Detecting changes in gene expression in untreated de novo patients with PD is important for understanding disease pathogenesis and for identifying biomarkers for preclinical stage of PD. In this study we investigate ST13 gene expression in the peripheral blood of different groups of patients with neurological diseases using reverse transcription reaction and real-time polymerase chain reaction (PCR). Our results suggest that the expression levels of ST13 cannot serve as a biomarker for early stages of PD.
Evidence for the presence of full-length PARK2 mRNA and Parkin protein in human blood
2009, Neuroscience Letters
Research on Parkinson's disease fails to pinpoint a single gene or a gene product as the causative factor. However, the early onset form of the disease may be caused by mutations in PARK2 gene. Some studies related to the biochemistry or other aspects of the PARK2 gene or its product mostly used cDNA generated from substantia nigra of the mid-brain. This is essentially because the presence of the 1.4 kb full-length PARK2 cDNA in human leukocytes is, so far, not demonstrated although some splice variants and short RT-PCR products were reported. In this study, we synthesized a 1.4 kb full-length PARK2 cDNA from human leukocytes, cloned and expressed it both in Escherichia coli and in HeLa cells. The presence of Parkin protein was also demonstrated in human serum using Western blotting and MALDI-TOF analysis. The results of this study showed a simple way for routine amplification of PARK2 cDNA from human blood and may become a useful diagnostic tool in the future.
Advanced microarray analysis highlights modified neuro-immune signaling in nucleated blood cells from Parkinson's disease patients
2008, Journal of Neuroimmunology
Laboratory tests for Parkinson's disease (PD) were recently extended to microarray analyses of nucleated blood cells. Here, we report the use of statistical and gene ontology tools to re-examine these microarray data. Distribution plots and PCA mapping enabled removal of several outliers out of the 105 analyzed PD and control samples, which improved the discriminative power for PD blood cells compared to healthy and neurological disease controls. Combined with gene ontology tests, our findings point at neuro-immune signaling-related transcripts as distinctly expressed early in PD progress and call for exploiting microarray tests also for follow-up of PD treatment efficacy.

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Differential expression of the parkin gene in the human brain and peripheral leukocytes

Abstract

Section snippets

Acknowledgements

Illegitimate transcription: application to the analysis of truncated transcripts of the dystrophin gene in non-muscle cultured cells from Duchenne and Becker patients

J. Clin. Invest.

Prevalence of Parkinson's disease in the elderly: the Rotterdam study

Neurology

A susceptibility locus for Parkinson's disease maps to chromosome 2p13

Nat. Genet.

Clinical analysis of 17 patients in 12 Japanese families with autosomal recessive type juvenile parkinsonism

Neurology