Papers by Martina Zizkova
Background β-catenin is a key component of Wnt signalling pathway which controls brain developmen... more Background β-catenin is a key component of Wnt signalling pathway which controls brain development and neurogenesis of the adult brain. Within a cell, β-catenin is continuously synthesised and its level is strictly regulated by phosphorylation and subsequent degradation via ubiquitin-proteasome system. Recently, the accumulation of β-catenin has been associated with Huntington disease as a consequence of the impaired degradation caused by mutated huntingtin. Aims Our study is aimed at monitoring β-catenin level in minipigs transgenic for the N-terminal part of human mutated huntingtin, the model developed in our institute. Methods Using Western blot and specific antibodies, transgenic animals and their siblings of the same age and the identical genetic background were examined for the expression of β-catenin. Furthermore, phosphorylation of β-catenin was monitored applying the method of phosphoprotein isolation. Finally, β-catenin was isolated by immunoprecipitation and subjected to...
Journal of Proteomics, 2015
Pluripotent stem cell-derived committed neural precursors are an important source of cells to tre... more Pluripotent stem cell-derived committed neural precursors are an important source of cells to treat neurodegenerative diseases including spinal cord injury. There remains an urgency to identify markers for monitoring of neural progenitor specificity, estimation of neural fate and follow-up correlation with therapeutic effect in preclinical studies using animal disease models. Cell surface capture technology was used to uncover the cell surface exposed N-glycoproteome of neural precursor cells upon neuronal differentiation as well as post-mitotic mature hNT neurons. The data presented depict an extensive study of surfaceome during neuronal differentiation, confirming glycosylation at a particular predicted site of many of the identified proteins. Quantitative changes detected in cell surface protein levels reveal a set of proteins that highlight the complexity of the neuronal differentiation process. Several of these proteins including the cell adhesion molecules ICAM1, CHL1, and astrotactin1 as well as LAMP1 were validated by SRM. Combination of immunofluorescence staining of ICAM1 and flow cytometry indicated a possible direction for future scrutiny of such proteins as targets for enrichment of the neuronal subpopulation from mixed cultures after differentiation of neural precursor cells. These surface proteins hold an important key for development of safe strategies in cell-replacement therapies of neuronal disorders. Neural stem and/or precursor cells have a great potential for cell-replacement therapies of neuronal diseases. Availability of well characterised and expandable neural cell lineage specific populations is critical for addressing such a challenge. In our study we identified and relatively quantified several hundred surface N-glycoproteins in the course of neuronal differentiation. We further confirmed the abundant changes for several cell adhesion proteins by SRM and outlined a strategy for utilisation of such N-glycoproteins in antibody based cell sorting. The comprehensive dataset presented here demonstrates the molecular background of neuronal differentiation highly useful for development of new plasma membrane markers to identify and select neuronal subpopulation from mixed neural cell cultures.
Expert Review of Proteomics, 2015
Neurodegenerative diseases are devastating disorders and the demands on their treatment are set t... more Neurodegenerative diseases are devastating disorders and the demands on their treatment are set to rise in connection with higher disease incidence. Knowledge of the spatiotemporal profile of cellular protein expression during neural differentiation and definition of a set of markers highly specific for targeted neural populations is a key challenge. Intracellular proteins may be utilized as a readout for follow-up transplantation and cell surface proteins may facilitate isolation of the cell subpopulations, while secreted proteins could help unravel intercellular communication and immunomodulation. This review summarizes the potential of proteomics in revealing molecular mechanisms underlying neural differentiation of stem cells and presents novel candidate proteins of neural subpopulations, where understanding of their functionality may accelerate transition to cell replacement therapies.
PROTEOMICS - Clinical Applications, 2014
Abbreviations: BACHD, bacterial arteficial chromosome; BDNF, brain-derived neurotrophic factor; C... more Abbreviations: BACHD, bacterial arteficial chromosome; BDNF, brain-derived neurotrophic factor; CAG, cytosine-adenine-guanine; CB1, cannabinoid receptor type 1; CNS, central nervous system; CREB, c-AMP resposive element -binding protein; CSF, cerebrospinal fluid; DARPP32, dopamine-and cyclic AMP-regulated phosphoprotein; Gnb2l1, Guanine nucleotide-binding protein subunit beta-2-like 1; H2AFY, H2A histone family member Y; HD, Huntington's disease; HD protein, huntingtin protein; HTT, huntingtin gene; iPSCs, induced pluripotent stem cell; MSNs, medium spiny neurons; MRI, magnetic resonance imaging; Myo5a, Unconventional myosin-Va; PET, positron emission tomography; PGC-1α, peroxisome proliferator-activated receptor-γ coactivator 1α; polyQ, polyglutamine; Prkra, Interferoninducible double-stranded RNA-dependent protein kinase activator A; REST, R element-1 silencing transcription factor; Rps6, ribosomal protein S6; SP1, specificity protein 1; Syt2, Synaptotagmin-2; TAFII130, transcription initiation factor TFIID subunit 4; TNFα, tumor necrosis factor α; UHDRS, Unified Huntington's Disease Rating Scale; YAC, yeast arteficial chromosome
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Papers by Martina Zizkova