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Single nuclei gene expression profile of brain cells from Pdgfbret/ret and Pdgfbret/wt mice

ABSTRACT: Brain calcification, the ectopic mineral deposits of calcium phosphate, is a common radiological finding in elderly and in various neurodegenerative disorders and a diagnostic criterion for primary familial brain calcification (PFBC). We made use of single nuclei RNA sequencing to understand the effect of calcification pathology in the Pdgfb (ret/ret) model of PFBC. Calcifications are located in deep brain regions - hypothalamus, thalamus, mid brain, pons of Pdgfb (ret/ret) mice and absent in control Pdgfb (ret/wt) mice. Forebrain cortex and hippocampus regions in both genotypes are calcification free. Detailed descriptions of the mouse model can be found in the following publications:

ORGANISM(S): Mus musculus

PROVIDER: GSE263392 | GEO | 2024/10/14

REPOSITORIES: GEO

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Project description:Genomic rearrangements leading to intragenic gene fusion are mainly found in some types of haematopoietic malignancies and sarcomas. Recently they have been described also in carcinomas such as the papillary thyroid histotype (60%-70%) and the Hürthle thyroid tumours (58%). The presence of junction oncogene constitutes an area of exciting research for emerging therapy as targeting the RET-PTC1 fusion oncogene by using small interfering RNA (siRNA) strategies since it is present only in the tumour cells and not in the surrounding normal cells. Therefore, we developed a siRNA against RET-PTC1 junction and assess its efficiency on the human papillary thyroid carcinoma cell line TPC-1 which spontaneously harbours the RET-PTC1 oncogene. The targeted genes are assessed by microarray analysis by comparing the regulated genes by the siRNA_RET-PTC1 vs a siRNA_RET developed on the RET part of the mRNA minus the siRNA_control that contain four mutation within the RET-PTC1 sequence. To test the targeted genes in the TPC-1 cell line that spontaneously harbours RET-PTC1 junction of two siRNAs developed: Within the RET-PTC1 junction, and in the mRNA RET part. A non-specific siRNA harbouring 4 mutations within the RET-PTC1 sequence was used as negative control (siRNA_control). By real-time PCR (Q-RT-PCR) we demonstrated that both siRNAs (siRNA_RET-PTC1 and siRNA_RET) significantly reduce RET mRNA levels of about 85 % in TPC-1 cells. The negative control did not show an effect of RET mRNA levels. Three independent transfections were performed on TPC-1 cells using 5 µl of Lipofectamine 2000 transfection reagent (Invitrogen, Cergy-Pontoise, France) and 50nM of i) siRNA_RET-PTC1 or ii) siRNA_RET or iii) siRNA_control that harbour 4 mutations within its sequence. Total RNAs of untreated cells and transfected cells were purified using the RNA cleanup and concentration kit (QIAGEN, Hilden, Germany) and gathered in 4 pools : 1) TPC-1 harbouring RET-PTC1 ; 2) TPC-1 silenced for RET-PTC1 with siRNA RET-PTC1 ; 3) TPC-1 silenced for siRNA RET ; 4) TPC-1 treated with the siRNA control.

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Single nuclei gene expression profile of brain cells from Pdgfbret/ret and Pdgfbret/wt mice

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