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1

Polymorphisms of the PARK2 gene and Parkin protein levels in patients with Parkinson’s disease

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Dorszewska J., Debek A., Oczkowska A., Florczak-Wyspianska J., Owecki M., Kozubski W.
Acta Neurobiologiae Experimentalis
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2013
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tom 73
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nr 1
Parkinson’s disease (PD) is one of the most common degenerative diseases of the extrapyramidal system, the frequency of which increases with age. It is now believed that the causes of PD are environmental and genetic factors. Important genetic factors resulting in PD are mutations in the PARK2 gene, which may affect the level of Parkin. The aim of the study was conducted on 234 individuals of the Polish population: 89 patients diagnosed with PD, 32 patients diagnosed with Parkinson’s syndrome and 113 individuals from the control group without neurological symptoms and characteristics of dementia. As a result of the methods of analysis demonstrated the following: G930C mutation of exon 8 in the PARK2 gene, which was analysed by performing PCR-RFLP. Detection of deletion of exon 2 using PCR. Whereas the evaluation of mutations within exon 11 in PARK2 gene was performed using HRM method and sequencing. Also performed to measure the concentration of Parkin’s plasma in blood using ELISA method. The study results no presence of the deletion of exon 2 in the PARK2 gene in any individual study. At the same time, it was almost 3-times higher frequency of G930C mutation in exon 8 PARK2 in patients with PD and almost 6-times higher incidence of mutation G1281 A in exon 11 of PARK2 in PD patients compared with controls. At the same time, in the present study demonstrated that the presence of mutations in 8 and 11 exon of PARK2 gene does not appear to be associated with the generate of Parkin’s plasma concentration. Genotype-fenotype study in the PARK2 gene can constitute intravital diagnostic tests in patients with PD, as well as in patients diagnosed with Parkinson’s syndrome in the course of a degenerative disease.
2

Elevated expression of alpha7 neuronal nicotinic acetylcholine receptor during the early stages of damage by oxidative stress in the aging rat brain

84%
Dorszewska J., Rozycka A., Helias-Rodzewicz Z., Kempisty B., Grzelak T., Ziemnicka K., Oczkowska A., Steinborn B., Jagodzinski P.
Acta Neurobiologiae Experimentalis
|
2013
|
tom 73
|
nr 1
Aging is accompanied by a high level of oxidized form of guanine, 8-oxo-2’deoxyguanosine (8-oxo-2’dG), and decreased level of 8-oxoguanine glycosylase 1 (OGG1) in the brain. The development and progression of neurodegenerative disorders are also characterized by dysfunction or loss of the brain nicotinic acetylcholine receptors (nAChRs). To study whether the differences in nAChRs expression in the rat brain occur due to aging or oxidative stress we analyzed RNA and protein levels of α7, α4 and β2 subunits by RQ-PCR and Western blot validation in three brain structures: cerebral grey matter (CGM), sub-cortical white matter (SCWM) and cerebellum (Ce) of twenty one female Wistar rats. The first group consisted of five 3.0–3.5-month-old females, which was assigned as a young control group. The remaining sixteen females aged of 18–24 month were divided into three following groups: (1) aged control group of 5 rats; (2) a vehicle group of 5 rats which received intraperitoneal injections of deionized water; (3) memantine-treated group of 6 rats. In each group, the selected brain areas have also been analyzed to determinate the levels of oxidative stress. In this study, age- and stress- dependent differential RNA and protein expression levels were approved only in OGG1 and α7 nAChR proteins. In all analyzed brain structures of young and old controls, the levels of oxidized form of guanine were similar. Stress relevant to water injection increased the level of 8-oxo-2’dG in the cerebellum of old control rats (Ce, P<0.05). The old controls demonstrated an important reduction of OGG1 mRNA expression in CGM and Ce regions compared to young individuals (CGM P=0.03; Ce P=0.2). Western blot analysis has also revealed a reduction of OGG1 protein in the sub-cortical white matter of old individuals (SCWM, P=0.03). However, there was no important influence of water administration on OGG1 expression in all brain regions. In all analyzed brain structures, expression of α7 nAChR was down-regulated in old controls compared to young controls. However, this decrease was only significant in SCWM area (SCWM, P<0.05). Treatment with H2O caused a significant increase in RNA and protein levels of α7 nAChR in SCWM as compared to this brain structure of the aged control rats (SCWM, P<0.01). Our results suggest that aging of the rat brain is mostly associated with decreased expression of OGG1 as well as with deficit of α7 nAChR in the sub-cortical white matter. Stress relevant to water injection increases the level of 8-oxo-2’dG in the aging rat brain, but clearly overcomes the α7 nAChR deficit. A significant increase of the α7 nAChR expression in the SCWM of H2O-treated rats suggests that these receptors play an important role in compensatory mechanisms facilitating the impaired cholinergic neurotransmission following oxidative stress in the aging rat brain.
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