1Department of Pathology, Hospital Universitari and Health Sciences Research Institute Germans Trias i Pujol, Universitat Autònoma de Barcelona, Spain 2Department of Neurology, Hospital Universitari de Bellvitge, L’Hospitalet de Llobregat, Spain 3Department of Neurology, Hospital Universitari Germans Trias i Pujol, Badalona, Barcelona, Spain.
Parkinson disease (PD) and dementia with Lewy bodies (DLB) are Lewy body diseases characterized by abnormal alpha-synuclein deposits and overlapping pathological features in the brain. Several studies have shown that glucocerebrosidase (GBA) deficiency is involved in the development of LB diseases. Here, we aimed to find out if this deficiency starts at the transcriptional level, also involves alternative splicing, and if GBA expression changes in brain are also detectable in blood of patients with LB diseases. The expression of three GBA transcript variants (GBAtv1, GBAtv2 and GBAtv5) was analyzed in samples from 20 DLB, 25 PD and 17 control brains and in blood of 20 DLB, 26 PD patients and 17 unaffected individuals. Relative mRNA expression was determined by real-time PCR. Expression changes were evaluated by the ΔΔCt method. In brain, specific expression profiles were identified in the temporal cortex of DLB and in the caudate nucleus of PD. In blood, significant GBA mRNA diminution was found in both DLB and PD patients. Early PD and early-onset DLB patients showed lowest GBA levels which were normal in PD patients with advanced disease and DLB patients who developed disease after 70 years of age. In conclusion, disease group specific GBA expression profiles were found in mostly affected areas of LBD. In blood, GBA expression was diminished in LB diseases, especially in patients with early onset DLB and in patients with early PD. Age of disease onset exerts an opposite effect on GBA expression in DLB and PD.
Perez-Roca Laia,Adame-Castillo Cristina,Campdelacreu Jaume, et al. Glucocerebrosidase mRNA is Diminished in Brain of Lewy Body Diseases and Changes with Disease Progression in Blood[J]. Aging and disease,
2018, 9(2): 208-219.
Table 1 Clinico-neuropathological characteristics of Lewy body disease cases and controls.
Name and NCBI1
Primer sequence (5’ - 3’)
ATC ACA TGA CCC ATC CAC A
ACT CAA AGG CTT GGG ACA T
TTC GCC GAC GTG GAT CCT CT
ACC GAG CTG TAG CCG AAG CT
TTC GCC GAC GAG ACT CTG GA
ACC TGA TGC CCA CGA CAC TG
TTC TCT TCG CCG ACG GTG CC
AGC TCC ATC CGT CGC CCA CT
ACG GGC ACA GGA ATC GGA TA
AAC TGC AGG GCT CGG TGA AT
TCT ACA ATG AGC TGC GTG TG
GGA TAG CAA CGT ACA TGG CT
AAC TGG GAC GAC ATG GAG AA
TAG ATG GGC ACA GTG TGG GT
ATG TGG TTG GAG AGC TCA TT
TGT CTC TGC CGA GTG AAG AT
ACA CAC AGC CTA CTT TCC AAG
TCA ATG TTG CCA CCA CAC TGT
Table 2 RNA primer sequences used for the amplification of GBA1 isoforms, beta-actin, GUS and PBGD.
Figure 1. Schematic representation of the five GBA transcripts and location of forward primers. Grey boxes represent exons and the lines, introns. Narrow red rectangles at the end of some exons indicate sequences, chosen for designing isoform-specific primers.
Figure 2. Relative GBA isoform expression in different brain areas. GBA expression in neural tissue estimated by appraising agarose gel electrophoretograms: tv, transcript variant; FC, frontal cortex; TC, temporal cortex; Ca, caudate nucleus; Put, putamen; NBM, Nucleus basalis of Meynert; Am, Amygdala; SN, Substantia nigra; Pt, pons; Cr, cerebellum. White fields correspond to lack of expression, light gray (1) to very slight expression, middle gray (2) to readily detectable expression, and dark gray (3) to high expression. The black fields represent very intense expression levels.
Figure 3. Expression profiles of GBA1 isoforms in three brain areas of LBD after adjustment with controls. The included areas were temporal cortex (TC) and caudate nucleus (Ca) from the groups of pure dementia with Lewy bodies (pDLB), common dementia with Lewy bodies (cDLB), Parkinson′s disease without dementia (PDND) and Parkinson′s disease with dementia (PDD). The results are shown as relative expression changes obtained by the ΔΔCt method in comparison with normal controls and are represented in a logarithmic scale. Grey areas represent normal expression range. * Significant expression change below 0.5.
Figure 4. GBA1tv1 expression in blood of DLB and PD patients in dependency on disease duration. GBA1tv1 expression was analyzed (A) in two groups and (B) for each patient individually. For (A), the results are shown as relative expression changes obtained by the ΔΔCt method in comparison with control individuals. * Significant expression change below 0.5. § Significant expression change between the disease duration subgroups. For (B) each point corresponds to the value of the expression change of each individual obtained by the ΔΔCt method, where ΔCt of patients was determined individually and ΔCt of control individuals was the mean value of the entire control group. Grey areas represent normal expression range.
age at onset (range)
male: female ratio
Disease duration since onset
age at onset (range)
age at onset (range)
male: female ratio
Disease duration since onset
age at onset (range)
male: female ratio
Table 3 Clinical characteristics of DLB and PD patients in the disease onset and duration groups.
Figure 5. GBA1tv1 expression in blood of DLB and PD patients in dependency on the age of disease onset. GBA1tv1 expression was analyzed (A) in two groups and (B) for each patient individually. For (A), the results are shown as relative expression changes obtained by the ΔΔCt method in comparison with control individuals. *Significant expression change below 0.5. #Significant expression change between the age-at-onset dependent subgroups. For (B) each point corresponds to the value of the expression change of each individual obtained by the ΔΔCt method, where ΔCt of patients was determined individually and ΔCt of control individuals was the mean value of the entire control group. Grey areas represent normal expression range.
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