Cortical glial cells contain both ionotropic and metabotropic glutamate receptors. Despite several efforts, a comprehensive analysis of the entire family of glutamate receptors and their subunits present in glial cells is still missing. Here, we provide an overall picture of the gene expression of ionotropic (AMPA, kainate, NMDA) and the main metabotropic glutamate receptors in cortical glial cells isolated from GFAP/EGFP mice before and after focal cerebral ischemia. Employing single-cell RT-qPCR, we detected the expression of genes encoding subunits of glutamate receptors in GFAP/EGFP-positive (GFAP/EGFP(+)) glial cells in the cortex of young adult mice. Most of the analyzed cells expressed mRNA for glutamate receptor subunits, the expression of which, in most cases, even increased after ischemic injury. Data analyses disclosed several classes of GFAP/EGFP(+) glial cells with respect to glutamate receptors and revealed in what manner their expression correlates with the expression of glial markers prior to and after ischemia. Furthermore, we also examined the protein expression and functional significance of NMDA receptors in glial cells. Immunohistochemical analyses of all seven NMDA receptor subunits provided direct evidence that the GluN3A subunit is present in GFAP/EGFP(+) glial cells and that its expression is increased after ischemia. In situ and in vitro Ca(2+) imaging revealed that Ca(2+) elevations evoked by the application of NMDA were diminished in GFAP/EGFP(+) glial cells following ischemia. Our results provide a comprehensive description of glutamate receptors in cortical GFAP/EGFP(+) glial cells and may serve as a basis for further research on glial cell physiology and pathophysiology.
- MeSH
- gliový fibrilární kyselý protein analýza biosyntéza MeSH
- glutamátové receptory analýza biosyntéza MeSH
- ischemie mozku metabolismus MeSH
- kultivované buňky MeSH
- lidé MeSH
- mozková kůra chemie metabolismus MeSH
- myši transgenní MeSH
- myši MeSH
- neuroglie chemie metabolismus MeSH
- receptory N-methyl-D-aspartátu analýza biosyntéza MeSH
- zelené fluorescenční proteiny analýza biosyntéza MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Two-Photon Processor (TPP) is a versatile, ready-to-use, and freely available software package in MATLAB to process data from in vivo two-photon calcium imaging. TPP includes routines to search for cell bodies in full-frame (Search for Neural Cells Accelerated; SeNeCA) and line-scan acquisition, routines for calcium signal calculations, filtering, spike-mining, and routines to construct parametric fields. Searching for somata in artificial in vivo data, our algorithm achieved better performance than human annotators. SeNeCA copes well with uneven background brightness and in-plane motion artifacts, the major problems in simple segmentation methods. In the fast mode, artificial in vivo images with a resolution of 256 × 256 pixels containing ≈ 100 neurons can be processed at a rate up to 175 frames per second (tested on Intel i7, 8 threads, magnetic hard disk drive). This speed of a segmentation algorithm could bring new possibilities into the field of in vivo optophysiology. With such a short latency (down to 5-6 ms on an ordinary personal computer) and using some contemporary optogenetic tools, it will allow experiments in which a control program can continuously evaluate the occurrence of a particular spatial pattern of activity (a possible correlate of memory or cognition) and subsequently inhibit/stimulate the entire area of the circuit or inhibit/stimulate a different part of the neuronal system. TPP will be freely available on our public web site. Similar all-in-one and freely available software has not yet been published.
- MeSH
- algoritmy * MeSH
- aniliny analýza MeSH
- fluoresceiny analýza MeSH
- fluorescenční barviva MeSH
- mikroskopie fluorescenční multifotonová metody MeSH
- mozková kůra chemie MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- software * MeSH
- vápník analýza MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The perirhinal cortex (PRC) composed of areas 35 and 36 forms an important route for activity transfer between the hippocampus-entorhinal cortex and neocortex. Its function in memory formation and consolidation as well as in the initiation and spreading of epileptic activity was already partially elucidated. We studied the general pattern of calretinin (CR), parvalbumin (PV) and calbindin (CB) immunoreactivity and its corrected relative optical density (cROD) as well as morphological features and density of CR and PV immunoreactive (CR+, PV+) interneurons in the rat PRC. Neighboring neocortical association area Te3V was analyzed as well. The PRC differed from the Te3V in higher CR and lower PV overall immunoreactivity level. On CR immunostained sections, the difference between high cROD value in area 35 and low cROD value in area Te3V reached statistical significance (p<0.05). The pattern of CB immunoreactivity was similar to that of the neocortex. Vertically oriented bipolar neurons were the most common morphological type of CR+ neurons, multipolar neuronal morphology was typical among PV+ neurons and vertically oriented bipolar neurons and multipolar neurons were approximately equally frequent among CB+ neurons. The density of CR+ and PV+ neurons was stereologically measured. While the density of PV+ neurons was not significantly different in PRC when compared to Te3V, density of CR+ neurons in area 35 was significantly higher by comparison with Te3V (p<0.05). Further, the overall neuronal density was measured on Nissl stained sections and the proportion of CR+ and PV+ interneurons was expressed as a percentage of the total neurons counts. The percentage of CR+ interneurons was higher in area 35 by comparison with area Te3 (p<0.05), while the percentage of PV+ interneurons did not significantly differ among the examined areas. In conclusion, the PRC possesses specific interneuronal equipment with unusually high proportion of CR+ interneurons, what might be of importance for the presumed gating function of PRC in normal and diseased states.
- MeSH
- interneurony chemie metabolismus MeSH
- krysa rodu rattus MeSH
- mozková kůra chemie metabolismus MeSH
- neurony chemie metabolismus MeSH
- parvalbuminy analýza metabolismus MeSH
- S100 kalcium vázající protein G analýza metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- kyseliny dokosahexaenové * MeSH
- lidé MeSH
- mozková kůra chemie MeSH
- regulace genové exprese MeSH
- Check Tag
- lidé MeSH
Immunohistochemical studies of the presence of lactosylceramide (LacCer) in lysosomal storage disorders (LSDs) were done using anti-LacCer monoclonal antibody of the CDw 17 type (clone MG-2). No sign of an association between LacCer and the lysosomal system in normal cells was observed, except for histiocytes active in phagocytosis. A comparative study of a group of LSDs showed a general tendency for LacCer to increase in storage cells in Niemann-Pick disease type C (NPC), and types A and B, GM1 gangliosidosis, acid lipase deficiency, glycogen storage disease type II and mucopolysaccharidoses. LacCer accumulated in storage cells despite normal activity of relevant lysosomal degrading enzymes. The accumulation of LacCer displayed variability within storage cell populations, and was mostly expressed in neurons in NPC. An absence of the increase in LacCer in storage cells above control levels was seen in neuronal ceroid lipofuscinoses (neurons and cardiocytes) and in Fabry disease. Gaucher and Krabbe cells showed significantly lower levels, or even the absence, of LacCer compared with control macrophages. Results of immunohistochemistry were corroborated by semiquantitative lipid thin-layer chromatography (TLC). It is suggested that different associations of LacCer with the lysosomal storage process may reflect differences in glycosphingolipid turnover induced by the storage-compromised lysosomal/endosomal system.
- MeSH
- biologické markery analýza MeSH
- CD antigeny analýza metabolismus MeSH
- chromatografie na tenké vrstvě metody MeSH
- dítě MeSH
- dospělí MeSH
- financování vládou MeSH
- histiocyty chemie metabolismus patologie MeSH
- imunohistochemie metody MeSH
- játra chemie metabolismus patologie MeSH
- laktosylceramidy analýza metabolismus MeSH
- lyzozomální nemoci z ukládání klasifikace metabolismus patologie MeSH
- makrofágy chemie metabolismus patologie MeSH
- mozková kůra chemie metabolismus patologie MeSH
- neurony chemie metabolismus patologie MeSH
- slezina chemie metabolismus patologie MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- Publikační typ
- srovnávací studie MeSH
- MeSH
- alkany chemie MeSH
- dehydroepiandrosteron analogy a deriváty izolace a purifikace škodlivé účinky MeSH
- dehydroepiandrosteronsulfát analýza izolace a purifikace MeSH
- krysa rodu rattus MeSH
- mozek MeSH
- mozková kůra chemie MeSH
- věkové faktory MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- MeSH
- biologické modely MeSH
- buněčná membrána chemie metabolismus účinky léků MeSH
- krysa rodu rattus MeSH
- lithium farmakologie krev MeSH
- mastné kyseliny analýza metabolismus MeSH
- mozková kůra chemie metabolismus účinky léků MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
- Klíčová slova
- PARTUSISTEN (BOEHRING),
- MeSH
- centrální nervový systém chemie MeSH
- chromatografie plynová MeSH
- krysa rodu rattus MeSH
- mastné kyseliny krev MeSH
- mozková kůra chemie MeSH
- těhotenství u zvířat účinky léků MeSH
- těhotenství MeSH
- tokolytika aplikace a dávkování farmakologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
- MeSH
- corpus striatum chemie metabolismus MeSH
- DNA biosyntéza MeSH
- hipokampus chemie metabolismus MeSH
- krysa rodu rattus MeSH
- mozková kůra chemie metabolismus MeSH
- reperfuze MeSH
- thymidin aplikace a dávkování MeSH
- tranzitorní ischemická ataka metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
