A5036213765- Neuroinflammation and Neurodegeneration Mechanisms
- MicroRNA in disease regulation
- Single-cell and spatial transcriptomics
- Cancer-related molecular mechanisms research
- Neurogenesis and neuroplasticity mechanisms
- Advanced biosensing and bioanalysis techniques
- Molecular Biology Techniques and Applications
- Alzheimer's disease research and treatments
- Extracellular vesicles in disease
- Immune cells in cancer
- RNA modifications and cancer
- Neuroscience and Neuropharmacology Research
- Protease and Inhibitor Mechanisms
- Photoacoustic and Ultrasonic Imaging
- Cell Adhesion Molecules Research
- Circular RNAs in diseases
- Biosensors and Analytical Detection
- Emergency and Acute Care Studies
- Epigenetics and DNA Methylation
- Mesenchymal stem cell research
- Trauma and Emergency Care Studies
- Neuroblastoma Research and Treatments
- Nanopore and Nanochannel Transport Studies
- Neurological Disease Mechanisms and Treatments
- Atherosclerosis and Cardiovascular Diseases
Ludwig-Maximilians-Universität München
2022-2024
Czech Academy of Sciences, Institute of Biotechnology
2016-2023
Czech Academy of Sciences
2016-2022
Palacký University Olomouc
2017-2022
Health Service Executive
2016
MicroRNAs are a class of small non-coding RNAs that serve as important regulators gene expression at the posttranscriptional level. They stable in body fluids and pose great potential to biomarkers. Here, we present highly specific, sensitive cost-effective system quantify miRNA based on two-step RT-qPCR with SYBR-green detection chemistry called Two-tailed RT-qPCR. It takes advantage novel, target-specific primers for reverse transcription composed two hemiprobes complementary different...
Ischemic stroke is a well-recognized disease of aging, yet it unclear how the age-dependent vulnerability occurs and what are underlying mechanisms. To address these issues, we perform comprehensive RNA-seq analysis ischemic stroke, their interaction in 3- 18-month-old mice. We assess differential gene expression across injury status age, estimate cell type proportion changes, assay results against range transcriptional signatures from literature, unsupervised co-expression analysis,...
Abstract Understanding the complexity of cellular function within a tissue necessitates combination multiple phenotypic readouts. Here, we developed method that links spatially-resolved gene expression single cells with their ultrastructural morphology by integrating multiplexed error-robust fluorescence in situ hybridization (MERFISH) and large area volume electron microscopy (EM) on adjacent sections. Using this method, characterized transcriptional responses glial infiltrating T-cells...
Abstract Age-related myelin damage induces inflammatory responses, yet its involvement in Alzheimer’s disease remains uncertain, despite age being a major risk factor. Using mouse model of disease, we found that amyloidosis itself triggers age-related oligodendrocyte and damage. Mechanistically, CD8 + T cells promote the progressive accumulation abnormally interferon-activated microglia display myelin-damaging activity. Thus, our data suggest immune responses against myelinating...
In this study, we investigated the influence of metformin (MF) on proliferation and viability adipose-derived stromal cells isolated from horses (EqASCs). We determined effect cell metabolism in terms mitochondrial oxidative status. Our purpose was to evaluate derived healthy (EqASCHE) individuals affected by equine metabolic syndrome (EqASCEMS). The were treated with 0.5 μM MF for 72 h. proliferative activity evaluated based measurement BrdU incorporation during DNA synthesis, as well...
Abstract BACKGROUND Recent advances allowing quantification of RNA from single cells are revolutionizing biology and medicine. Currently, almost all single-cell transcriptomic protocols rely on reverse transcription (RT). However, RT is recognized as a known source variability, particularly with low amounts RNA. Recently, several new transcriptases (RTases) the potential to decrease loss information have been developed, but knowledge their performance limited. METHODS We compared 11 RTases...
NG2 cells, a fourth glial cell type in the adult mammalian central nervous system, produce oligodendrocytes healthy tissue, and display wide differentiation potential under pathological conditions, where they could give rise to reactive astrocytes. The factors that control of cells after focal cerebral ischemia (FCI) are largely unknown. Here, we used transgenic Cspg4‐cre/Esr1/ROSA26Sortm14(CAG‐tdTomato) mice, which tamoxifen administration triggers expression red fluorescent protein...
Abstract Circulating cell-free microRNAs are promising candidates for minimally invasive clinical biomarkers the diagnosis, prognosis and monitoring of many human diseases. Despite substantial efforts invested in field, research so far has failed to deliver expected results. One contributing factors is general lack agreement between various studies, partly due considerable technical challenges accompanying workflow. Pre-analytical variables including sample collection, RNA isolation,...
Abstract NG2 cells represent precursors of oligodendrocytes under physiological conditions; however, following cerebral ischemia they play an important role in glial scar formation. Here, we compared the expression profiles oligodendroglial lineage cells, after focal (FCI) and Alzheimer's‐like pathology using transgenic mice, which enables genetic fate‐mapping Cspg4‐positive their progeny, based on red fluorescent protein tdTomato. tdTomato‐positive possessed profile oligodendrocytes; cell...
Modification of the extracellular matrix (ECM) is one major processes in pathology brain damage following an ischemic stroke. However, our understanding how age-related ECM alterations may affect stroke pathophysiology and its outcome still very limited.We conducted ECM-targeted re-analysis previously obtained RNA-Seq dataset aging, their interactions young adult (3-month-old) aged (18-month-old) mice. The permanent middle cerebral artery occlusion (pMCAo) rodents was used as a model...
In this study, we aimed to disclose the impact of amyloid-β toxicity and tau pathology on astrocyte swelling, their volume recovery extracellular space (ECS) diffusion parameters, namely fraction (α) tortuosity (λ), in a triple transgenic mouse model Alzheimer's disease (3xTg-AD). Astrocyte changes, which reflect ability take up ions/neurotransmitters, were quantified during after exposure hypo-osmotic stress, or hyperkalemia acute hippocampal slices, correlated with alterations ECS...
Abstract Current spatial transcriptomics methods identify cell states in a context but lack morphological information. Scanning electron microscopy, contrast, provides structural details at nanometer resolution lacks molecular decoding of the diverse cellular states. To address this, we correlated MERFISH with large area volume microscopy using adjacent tissue sections. We applied our technology to characterize damage-associated microglial identities mouse brain, allowing us, for first time,...
Small RNA-sequencing (RNA-Seq) is being increasingly used for profiling of circulating microRNAs (miRNAs), a new group promising biomarkers. Unfortunately, small RNA-Seq protocols are prone to biases limiting quantification accuracy, which motivated development several novel methods. Here, we present comparison all library preparation approaches that commercially available miRNAs in biofluids. Using synthetic and human plasma samples, compared performance traditional two-adaptor ligation...
ABSTRACT Background Recent technical advances allowing quantification of RNA from single cells are revolutionizing biology and medicine. Currently, almost all single-cell transcriptomic protocols rely on conversion to cDNA by reverse transcription (RT). However, RT is recognized as highly limiting step due its inherent variability suboptimal sensitivity, especially at minute amounts RNA. Primary factor influencing outcome transcriptase (RTase). Recently, several new RTases with potential...
Abstract Ischemic stroke is one of the leading causes mortality and major healthcare economic burden. It a well-recognized disease aging, yet it unclear how age-dependent vulnerability occurs what are underlying mechanisms. To address these issues, we performed comprehensive RNA-Seq analysis ischemic their interaction using model permanent middle cerebral artery occlusion (MCAO) in 3 18 month old female mice. We assessed differential gene expression across injury status age, estimated cell...