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Proteomic analysis of cerebellar synaptosomes in presymptomatic cystatin B –deficient mice

ABSTRACT: Loss-of-function mutations in cystatin B (CSTB) cause progressive myoclonus epilepsy of Unverricht-Lundborg type (EPM1). Cstb-deficiency in mice leads to early alterations in GABAergic signaling, and causes neuroinflammation followed by progressive neurodegeneration, manifesting as progressive myoclonus and ataxia. The proteome of cerebellar synaptosomes of presymptomatic Cstb-/- mice were characterized by LC-ESI-MS/MS to gain insight into disease formation and progression.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Brain

SUBMITTER: Tuula Nyman

LAB HEAD: Tuula Nyman

PROVIDER: PXD019370 | Pride | 2020-10-20

REPOSITORIES: Pride

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Proteome analysis of mouse cerebellar synaptosomes

Project description:In this study we have compared the proteomes from cerebellar synaptosome of early symptomatic Cstb-/- and wild type mice to gain insight into disease onset and progression of cystatin B deficiency. Biallelic loss-of-function mutations in the CSTB gene cause progressive myoclonus epilepsy of Unverricht-Lundborg type (EPM1). In the mouse model of EPM1, CSTB deficiency manifests from one month of age as progressive neurodegeneration, myoclonus and ataxia, which is preceded by neuroinflammation, alterations in GABAergic signaling, and a widespread rearrangement of the mitochondrial proteome in synapses.

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Project description:Progressive myoclonus epilepsy of Unverricht-Lundborg type (EPM1) is an inherited neurodegenerative disease with myoclonus, seizures and ataxia, caused by the mutations in cystatin B (CSTB) gene. In an approach towards understanding the molecular basis of pathogenic events in EPM1 we have utilized the cystatin B deficient mice (Cstb-/-), a model for the disease. We have characterized the gene expression changes from the cerebellum of Cstb-/- mouse at postnatal day 7 (P7) and P30 as well as in cultured cerebellar granule cells using a pathway-based approach. A marked upregulation of immune response genes was seen at P30, reflecting the ongoing neuropathology, however, the observed alterations in complement cascade genes could also imply defects in synaptic plasticity. Differentially expressed genes in pre-symptomatic Cstb-/- animals at P7 were connected to synaptic function and plasticity and in cultured cerebellar granule cells to cellular biogenesis, cytoskeleton and intracellular transport. Especially GABAergic pathways were affected. The sample preparation and hybridzation of each cRNAs on GeneChipM-BM-. Mouse Genome 430 2.0 arrays (Affymetrix, Santa Clara, CA, USA) were performed in Helsinki Biomedicum Biochip Center (Finland).

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