AIM: Astrocytes respond to stressors by acquiring a reactive state characterized by changes in their morphology and function. Molecules underlying reactive astrogliosis, however, remain largely unknown. Given that several studies observed increase in the Amyloid Precursor Protein (APP) in reactive astrocytes, we here test whether APP plays a role in reactive astrogliosis. METHODS: We investigated whether APP instigates reactive astroglios by examining in vitro and in vivo the morphology and function of naive and APP-deficient astrocytes in response to APP and well-established stressors. RESULTS: Overexpression of APP in cultured astrocytes led to remodeling of the intermediate filament network, enhancement of cytokine production, and activation of cellular programs centered around the interferon (IFN) pathway, all signs of reactive astrogliosis. Conversely, APP deletion abrogated remodeling of the intermediate filament network and blunted expression of IFN-stimulated gene products in response to lipopolysaccharide. Following traumatic brain injury (TBI), mouse reactive astrocytes also exhibited an association between APP and IFN, while APP deletion curbed the increase in glial fibrillary acidic protein observed canonically in astrocytes in response to TBI. CONCLUSIONS: The APP thus represents a candidate molecular inducer and regulator of reactive astrogliosis. This finding has implications for understanding pathophysiology of neurodegenerative and other diseases of the nervous system characterized by reactive astrogliosis and opens potential new therapeutic avenues targeting APP and its pathways to modulate reactive astrogliosis.

• Klíčová slova
• amyloid precursor protein, astrocytes, interferon pathway, lipopolysaccharide, reactive astrogliosis, traumatic brain injury,
• MeSH
• amyloidový prekurzorový protein beta * metabolismus   genetika   MeSH
• astrocyty * metabolismus   patologie   MeSH
• glióza * metabolismus   patologie   MeSH
• kultivované buňky MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši MeSH
• traumatické poranění mozku metabolismus   patologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• amyloidový prekurzorový protein beta * MeSH

The glia limitans superficialis (GLS) on the rodent cortical surface consists of astrocyte bodies intermingled with their cytoplasmic processes. Many studies have observed astrocyte reactivity in the medial prefrontal cortex (mPFC) parenchyma induced by a peripheral nerve injury, while the response of GLS astrocytes is still not fully understood. The aim of our study was to identify the reactivity of rat GLS astrocytes in response to sciatic nerve compression (SNC) over different time periods. The alteration of GLS astrocyte reactivity was monitored using immunofluorescence (IF) intensities of glial fibrillary acidic protein (GFAP), glutamine synthetase (GS), and NFκBp65. Our results demonstrated that SNC induced GLS astrocyte reactivity seen as increased intensities of GFAP-IF, and longer extensions of cytoplasmic processes into lamina I. First significant increase of GFAP-IF was observed on post-operation day 7 (POD7) after SNC with further increases on POD14 and POD21. In contrast, dynamic alteration of the extension of cytoplasmic processes into lamina I was detected as early as POD1 and continued throughout the monitored survival periods of both sham and SNC operations. The reactivity of GLS astrocytes was not associated with their proliferation. In addition, GLS astrocytes also displayed a significant decrease in GS immunofluorescence (GS-IF) and NFκB immunofluorescence (NFκB-IF) in response to sham and SNC operation compared with naïve control rats. These results suggest that damaged peripheral tissues (following sham operation as well as peripheral nerve lesions) may induce significant changes in GLS astrocyte reactivity. The signaling mechanism from injured peripheral tissue and nerve remains to be elucidated.

• Klíčová slova
• Glial fibrillary acidic protein, Glutamine synthetase, NFκBp65, Peripheral nerve lesion, Reactive astrocytes,
• MeSH
• astrocyty * metabolismus   patologie   MeSH
• gliový fibrilární kyselý protein metabolismus   MeSH
• krysa rodu Rattus MeSH
• nervus ischiadicus zranění   metabolismus   MeSH
• poranění periferního nervu * metabolismus   MeSH
• prefrontální mozková kůra metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• gliový fibrilární kyselý protein MeSH

Histone H3.3 glycine 34 to arginine/valine (G34R/V) mutations drive deadly gliomas and show exquisite regional and temporal specificity, suggesting a developmental context permissive to their effects. Here we show that 50% of G34R/V tumors (n = 95) bear activating PDGFRA mutations that display strong selection pressure at recurrence. Although considered gliomas, G34R/V tumors actually arise in GSX2/DLX-expressing interneuron progenitors, where G34R/V mutations impair neuronal differentiation. The lineage of origin may facilitate PDGFRA co-option through a chromatin loop connecting PDGFRA to GSX2 regulatory elements, promoting PDGFRA overexpression and mutation. At the single-cell level, G34R/V tumors harbor dual neuronal/astroglial identity and lack oligodendroglial programs, actively repressed by GSX2/DLX-mediated cell fate specification. G34R/V may become dispensable for tumor maintenance, whereas mutant-PDGFRA is potently oncogenic. Collectively, our results open novel research avenues in deadly tumors. G34R/V gliomas are neuronal malignancies where interneuron progenitors are stalled in differentiation by G34R/V mutations and malignant gliogenesis is promoted by co-option of a potentially targetable pathway, PDGFRA signaling.

• Klíčová slova
• GSX2, H3.3 G34R/V, PDGFRA, cell-of-origin, chromatin conformation, gliomas, interneuron progenitors, oncohistones, pediatric cancer, single-cell transcriptome,
• MeSH
• astrocyty metabolismus   patologie   MeSH
• biologické modely MeSH
• buněčný rodokmen MeSH
• chromatin metabolismus   MeSH
• embryo savčí metabolismus   MeSH
• epigeneze genetická MeSH
• genetická transkripce MeSH
• gliom genetika   patologie   MeSH
• histony genetika   metabolismus   MeSH
• interneurony metabolismus   MeSH
• karcinogeneze genetika   patologie   MeSH
• lysin metabolismus   MeSH
• mutace genetika   MeSH
• myši inbrední C57BL MeSH
• nádory mozku genetika   patologie   MeSH
• nervové kmenové buňky metabolismus   MeSH
• oligodendroglie metabolismus   MeSH
• přední mozek embryologie   MeSH
• přeprogramování buněk genetika   MeSH
• promotorové oblasti (genetika) genetika   MeSH
• regulace genové exprese u nádorů MeSH
• růstový faktor odvozený z trombocytů - receptor alfa genetika   metabolismus   MeSH
• stupeň nádoru MeSH
• transkriptom genetika   MeSH
• umlčování genů MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• chromatin MeSH
• histony MeSH
• lysin MeSH
• růstový faktor odvozený z trombocytů - receptor alfa MeSH

X-linked Adrenoleukodystrophy (X-ALD) is caused by mutations in the ABCD1 gene resulting in the accumulation of very long chain fatty acids (VLCFA). X-ALD is the most common peroxisomal disorder with adult patients (male and female) presenting with progressive spastic paraparesis with bladder disturbance, sensory ataxia with impaired vibration sense, and leg pain. 80% of male X-ALD patients have an adrenal failure, while adrenal dysfunction is rare in women with X-ALD. The objective of this study was to define optimal serum VLCFA cutoff values in patients with X-ALD-like phenotypes for the differentiation of genetically confirmed X-ALD and Non-X-ALD individuals. Three groups were included into this study: a) X-ALD cases with confirmed ABCD1 mutations (n = 34) and two Non-X-ALD cohorts: b) Patients with abnormal serum VCLFA levels despite negative testing for ABCD1 mutations (n = 15) resulting from a total of 1,953 VLCFA tests c) Phenotypically matching patients as Non-X-ALD controls (n = 104). Receiver operating curve analysis was used to optimize VLCFA cutoff values, which differentiate patients with genetically confirmed X-ALD and Non-X-ALD individuals. The serum concentration of C26:0 was superior to C24:0 for the detection of X-ALD. The best differentiation of Non-X-ALD and X-ALD individuals was obtained with a cutoff value of < 1.0 for the C24:0/C22:0 ratio resulting in a sensitivity of 97%, a specificity of 94.1% and a positive predictive value (PPV) of 83.8% for true X-ALD. Our findings further suggested a cutoff of < 0.02 for the ratio C26:0/C22:0 leading to a sensitivity of 90.9%, a specificity of 95.0%, and a PPV of 80.6%. Pearson correlation indicated a significant positive association between total blood cholesterol and VLCFA values. Usage of serum VLCFA are economical and established biomarkers suitable for the guidance of genetic testing matching the X-ALD phenotype. We suggest using our new optimized cutoff values, especially the two ratios (C24:0/C22:0 and C26:0/C22:0), in combination with standard lipid profiles.

• MeSH
• ABC transportéry genetika   MeSH
• adrenoleukodystrofie krev   diagnóza   MeSH
• astrocyty patologie   MeSH
• biologické markery krev   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mastné kyseliny krev   MeSH
• mutace genetika   MeSH
• senioři MeSH
• senzitivita a specificita MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ABC transportéry MeSH
• biologické markery MeSH
• mastné kyseliny MeSH

In this study, we focused on age-related changes in astrocyte functioning, predominantly on the ability of astrocytes to regulate their volume in response to a pathological stimulus, namely extracellular 50 mM K+ concentration. The aim of our project was to identify changes in the expression and function of transport proteins in the astrocytic membrane and properties of the extracellular space, triggered by aging. We used three-dimensional confocal morphometry, gene expression profiling, immunohistochemical analysis, and diffusion measurement in the hippocampal slices from 3-, 9-, 12-, and 18-month-old mice, in which astrocytes are visualized by enhanced green fluorescent protein under the control of the promoter for human glial fibrillary acidic protein. Combining a pharmacological approach and the quantification of astrocyte volume changes evoked by hyperkalemia, we found that marked diversity in the extent of astrocyte swelling in the hippocampus during aging is due to the gradually declining participation of Na+-K+-Cl- transporters, glutamate transporters (glutamate aspartate transporter and glutamate transporter 1), and volume-regulated anion channels. Interestingly, there was a redistribution of Na+-K+-Cl- cotransporter and glutamate transporters from astrocytic soma to processes. In addition, immunohistochemical analysis confirmed an age-dependent decrease in the content of Na+-K+-Cl- cotransporter in astrocytes. The overall extracellular volume changes revealed a similar age-dependent diversity during hyperkalemia as observed in astrocytes. In addition, the recovery of the extracellular space was markedly impaired in aged animals.

• Klíčová slova
• Aging, Astrocyte, Hippocampus, Potassium/glutamate uptake, Volume regulation,
• MeSH
• astrocyty účinky léků   metabolismus   patologie   fyziologie   MeSH
• draslík farmakologie   MeSH
• gliový fibrilární kyselý protein MeSH
• hipokampus cytologie   MeSH
• myši transgenní MeSH
• stárnutí patologie   fyziologie   MeSH
• velikost buňky * MeSH
• zelené fluorescenční proteiny MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• draslík MeSH
• gliový fibrilární kyselý protein MeSH
• zelené fluorescenční proteiny MeSH

Tuberous sclerosis complex (TSC) is a genetic disorder characterized by frequent noncancerous neoplasia in the brain, which can induce a range of severe neuropsychiatric symptoms in humans, resulting from out of control tissue growth. The causative spontaneous loss-of-function mutations have been also identified in rats. Herein, we studied histopathological and molecular changes in brain lesions of the Eker rat model carrying germline mutation of the tsc2 gene, predisposed to multiple neoplasias. Predominant subcortical tumors were analyzed, along with a rare form occurring within the pyriform lobe. The uniform composition of lesions supports the histochemical parity of malformations, with immunofluorescence data supporting their neuro-glial origin. Massive depletion of mature neurons and axonal loss were evident within lesions, with occasional necrotic foci implying advanced stage of pathology. Enrichment of mesenchymal-derived cell markers with hallmarks of neurogenesis and active microglia imply enhanced cell proliferation, with local immune response. The depletion of capillaries within the core was complemented by the formation of dense mesh of nascent vessels at the interface of neoplasia with healthy tissue, implying large-scale vascular remodeling. Taken as a whole, these findings present several novel features of brain tumors in Eker rat model, rendering it suitable for studies of the pathobiology and progression of primary brain tumors, with therapeutic interventions.

• Klíčová slova
• Eker rat, Glioma, Hamartomas, Microglial activation, Nascent capillaries, Neuro-oncology, Tuberous sclerosis complex,
• MeSH
• astrocyty patologie   MeSH
• axony patologie   MeSH
• mikroglie patologie   MeSH
• mozek krevní zásobení   patologie   MeSH
• nádory mozku krevní zásobení   etiologie   patologie   MeSH
• neurony patologie   MeSH
• potkani Long-Evans MeSH
• remodelace cév * MeSH
• tuberin genetika   MeSH
• tuberózní skleróza komplikace   patologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• Tsc2 protein, rat MeSH Prohlížeč
• tuberin MeSH

Transient receptor potential vanilloid type 4 (TRPV4) channels are involved in astrocyte volume regulation; however, only limited data exist about its mechanism in astrocytes in situ. We performed middle cerebral artery occlusion in adult mice, where we found twice larger edema 1 day after the insult in trpv4-/- mice compared to the controls, which was quantified using magnetic resonance imaging. This result suggests disrupted volume regulation in the brain cells in trpv4-/- mice leading to increased edema formation. The aim of our study was to elucidate whether TRPV4 channel-based volume regulation occurs in astrocytes in situ and whether the disrupted volume regulation in trpv4-/- mice might lead to higher edema formation after brain ischemia. For our experiments, we used trpv4-/- mice crossed with transgenic mice expressing enhanced green fluorescent protein (EGFP) under the control of the glial fibrillary acidic protein promoter, which leads to astrocyte visualization by EGFP expression. For quantification of astrocyte volume changes, we used two-dimensional (2D) and three-dimensional (3D) morphometrical approaches and a quantification algorithm based on fluorescence intensity changes during volume alterations induced by hypotonicity or by oxygen-glucose deprivation. In contrast to in vitro experiments, we found little evidence of the contribution of TRPV4 channels to volume regulation in astrocytes in situ in adult mice. Moreover, we only found a rare expression of TRPV4 channels in adult mouse astrocytes. Our data suggest that TRPV4 channels are not involved in astrocyte volume regulation in situ; however, they play a protective role during the ischemia-induced brain edema formation.

• Klíčová slova
• TRPV4 channels, astrocytes, ischemia, volume regulation,
• MeSH
• astrocyty metabolismus   patologie   MeSH
• edém mozku etiologie   metabolismus   patologie   MeSH
• infarkt arteria cerebri media metabolismus   patologie   MeSH
• ischemie mozku komplikace   MeSH
• kationtové kanály TRPV genetika   metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• primární buněčná kultura MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kationtové kanály TRPV MeSH
• Trpv4 protein, mouse MeSH Prohlížeč

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 of oligodendroglial lineage cells, after focal cerebral ischemia (FCI) and in Alzheimer's-like pathology using transgenic mice, which enables genetic fate-mapping of Cspg4-positive NG2 cells and their progeny, based on the expression of red fluorescent protein tdTomato. tdTomato-positive cells possessed the expression profile of NG2 cells and oligodendrocytes; however, based on the expression of cell type-specific genes, we were able to distinguish between them. To shed light on the changes in the expression patterns caused by FCI, we employed self-organizing Kohonen maps, enabling the division of NG2 cells and oligodendrocytes into subpopulations based on similarities in the expression profiles of individual cells. We identified three subpopulations of NG2 cells emerging after FCI: proliferative; astrocyte-like and oligodendrocyte-like NG2 cells; such phenotypes were further confirmed by immunohistochemistry. Oligodendrocytes themselves formed four subpopulations, which reflected the process of oligodendrocytes maturation. Finally, we used 5-ethynyl-2' deoxyuridine (EdU) labeling to reveal that NG2 cells can differentiate directly into reactive astrocytes without preceding proliferation. In contrast, in Alzheimer's-like pathology we failed to identify these subpopulations. Collectively, here we identified several yet unknown differences between the expression profiles of NG2 cells and oligodendrocytes, and characterized specific genes contributing to oligodendrocyte maturation and phenotypical changes of NG2 cells after FCI. Moreover, our results suggest that, unlike in Alzheimer's-like pathology, NG2 cells acquire a multipotent phenotype following FCI.

• Klíčová slova
• Alzheimer's, NG2 cells, astrocytes, cerebral ischemia, oligodendrocytes,
• MeSH
• analýza jednotlivých buněk MeSH
• astrocyty patologie   fyziologie   MeSH
• ischemie mozku patologie   patofyziologie   MeSH
• modely nemocí na zvířatech MeSH
• mozek patologie   patofyziologie   MeSH
• myši transgenní MeSH
• prekurzorové buňky oligodendrocytů patologie   fyziologie   MeSH
• proliferace buněk fyziologie   MeSH
• regenerace nervu fyziologie   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Mucopolysaccharidosis type II (MPSII) is a rare X-linked lysosomal storage disorder caused by mutations in the iduronate-2-sulfatase (IDS) gene (IDS, Xq28). MPSII is characterized by skeletal deformities, hearing loss, airway obstruction, hepatosplenomegaly, cardiac valvular disease, and progressive neurological impairment. At the cellular level, IDS deficiency leads to lysosomal storage of glycosaminoglycans (GAGs), dominated by accumulation of dermatan and heparan sulfates. Human induced pluripotent stem cells (iPSC) represent an alternative system that complements the available MPSII murine model. Herein we report on the reprogramming of peripheral white blood cells from male and female MPSII patients into iPSC using a non-integrating protocol based on the Sendai virus vector system. We differentiated the iPSC lines into IDS deficient and GAG accumulating β-Tubulin III+ neurons, GFAP+ astrocytes, and CNPase+ oligodendrocytes. The lysosomal system in these cells displayed structural abnormalities reminiscent of those previously found in patient tissues and murine IDS deficient neuronal stem cells. Furthermore, quantitative determination of GAGs revealed a moderate increase in GAG levels in IDS deficient neurons and glia. We also tested the effects of recombinant IDS and found that the exogenous enzyme was internalized from the culture media and partially decreased the intracellular GAG levels in iPSC-derived neural cells; however, it failed to completely prevent accumulation of GAGs. In summary, we demonstrate that this human iPSC based model expresses the cellular and biochemical features of MPSII, and thus represents a useful experimental tool for further pathogenesis studies as well as therapy development and testing.

• MeSH
• astrocyty enzymologie   patologie   MeSH
• buněčný rodokmen MeSH
• fenotyp MeSH
• glykosaminoglykany metabolismus   MeSH
• iduronátsulfatasa genetika   metabolismus   MeSH
• indukované pluripotentní kmenové buňky enzymologie   patologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• lyzozomy enzymologie   patologie   MeSH
• mukopolysacharidóza II enzymologie   genetika   patologie   MeSH
• nervové kmenové buňky enzymologie   patologie   MeSH
• neurogeneze * MeSH
• neuroglie enzymologie   patologie   MeSH
• neurony enzymologie   patologie   MeSH
• oligodendroglie enzymologie   patologie   MeSH
• prekurzorové buňky oligodendrocytů enzymologie   patologie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• glykosaminoglykany MeSH
• iduronátsulfatasa MeSH

Aging-related tau astrogliopathy (ARTAG) is a recently introduced terminology. To facilitate the consistent identification of ARTAG and to distinguish it from astroglial tau pathologies observed in the primary frontotemporal lobar degeneration tauopathies we evaluated how consistently neuropathologists recognize (1) different astroglial tau immunoreactivities, including those of ARTAG and those associated with primary tauopathies (Study 1); (2) ARTAG types (Study 2A); and (3) ARTAG severity (Study 2B). Microphotographs and scanned sections immunostained for phosphorylated tau (AT8) were made available for download and preview. Percentage of agreement and kappa values with 95% confidence interval (CI) were calculated for each evaluation. The overall agreement for Study 1 was >60% with a kappa value of 0.55 (95% CI 0.433-0.645). Moderate agreement (>90%, kappa 0.48, 95% CI 0.457-0.900) was reached in Study 2A for the identification of ARTAG pathology for each ARTAG subtype (kappa 0.37-0.72), whereas fair agreement (kappa 0.40, 95% CI 0.341-0.445) was reached for the evaluation of ARTAG severity. The overall assessment of ARTAG showed moderate agreement (kappa 0.60, 95% CI 0.534-0.653) among raters. Our study supports the application of the current harmonized evaluation strategy for ARTAG with a slight modification of the evaluation of its severity.

• Klíčová slova
• ARTAG, Aging, Digital pathology, Interrater agreement, Neuropathology, Tau, Tau-astrogliopathy,
• MeSH
• astrocyty metabolismus   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• proteiny tau metabolismus   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• stárnutí patologie   MeSH
• stupeň závažnosti nemoci MeSH
• tauopatie patologie   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• proteiny tau MeSH