Recent work has emphasized the role of adhesion molecules in synaptic plasticity, including long-term potentiation in the hippocampus. Such adhesion molecules are concentrated in junctions that are characterized by dense thickenings on both sides of the junction and are called puncta adhaerentia (PA). Reconstruction from serial electron microscopy was used to determine the location and size of PA in the stratum radiatum of hippocampal area CA1, where many of the previous functional studies have been performed. PAs were found at the edges of synapses on 33% of dendritic spines. The areas occupied by PA were variable across different types of synapses, occupying 0.010+/-0.005 microm2 at macular synapses and 0.034+/-0.031 microm2 at perforated synapses. Another zone, called a vesicle-free transition zone (VFTZ), was identified. Like the PA, this zone also had no presynaptic vesicles and was located at the edges of synapses; however, unlike the PA, the presynaptic thickening was less than the postsynaptic thickening. Together, 45% of spine synapses had PA and/or VFTZ occupying 23+/-11% of the total junctional area between axons and spines. PA also occurred at nonsynaptic sites involving neuronal as well as glial elements. Most (64%) of these PAs occurred between nonsynaptic portions of dendritic spines and neighboring astrocytic processes. Smooth endoplasmic reticulum was often apposed to one or both sides of the synaptic and the nonsynaptic PA. These findings provide further data as a structural basis for understanding the roles of cell adhesion junctions in hippocampal synaptic function and plasticity.

• MeSH
• buněčná adheze fyziologie   MeSH
• dendrity ultrastruktura   MeSH
• elektronová mikroskopie MeSH
• hipokampus cytologie   MeSH
• hladké endoplazmatické retikulum ultrastruktura   MeSH
• inbrední kmeny potkanů fyziologie   MeSH
• krysa rodu Rattus MeSH
• mezibuněčné spoje fyziologie   ultrastruktura   MeSH
• neurony cytologie   ultrastruktura   MeSH
• neuropil ultrastruktura   MeSH
• počítačové zpracování obrazu MeSH
• synapse fyziologie   ultrastruktura   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Research Support, U.S. Gov't, P.H.S. MeSH

Tumor cells interact with blood constituents and these interactions promote metastasis. Selectins are vascular receptors facilitating interactions of tumor cells with platelets, leukocytes, and endothelium, but the role of endothelial E-selectin remains unclear. Here we show that E-selectin is a major receptor for monocyte recruitment to tumor cell-activated endothelium. Experimental and spontaneous lung metastasis using murine tumor cells, without E-selectin ligands, were attenuated in E-selectin-deficient mice. Tumor cell-derived CCL2 promoted endothelial activation, resulting in enhanced endothelial E-selectin expression. The recruitment of inflammatory monocytes to metastasizing tumor cells was dependent on the local endothelial activation and the presence of E-selectin. Monocytes promoted transendothelial migration of tumor cells through the induction of E-selectin-dependent endothelial retractions and a subsequent modulation of tight junctions through dephosphorylation of VE-cadherin. Thus, endothelial E-selectin shapes the tumor microenvironment through the recruitment, adhesion, and activation of monocytes that facilitate tumor cell extravasation and thereby metastasis. These findings provide evidence that endothelial E-selectin is a novel factor contributing to endothelial retraction required for efficient lung metastasis. Cancer Res; 76(18); 5302-12. ©2016 AACR.

• MeSH
• CD antigeny metabolismus   MeSH
• cévní endotel metabolismus   MeSH
• E-selektin metabolismus   MeSH
• endoteliální buňky metabolismus   MeSH
• imunoblotting MeSH
• imunoprecipitace MeSH
• invazivní růst nádoru patologie   MeSH
• kadheriny metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• monocyty metabolismus   MeSH
• mutantní kmeny myší MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• polymerázová řetězová reakce MeSH
• průtoková cytometrie MeSH
• těsný spoj metabolismus   patologie   MeSH
• transendoteliální a transepiteliální migrace fyziologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• cadherin 5 MeSH Prohlížeč
• CD antigeny MeSH
• E-selektin MeSH
• kadheriny MeSH

Destruction or dysfunction of limbal epithelial stem cells (LESCs) leads to unilateral or bilateral limbal stem cell deficiency (LSCD). Fifteen years have passed since the first transplantation of ex vivo cultivated oral mucosal epithelial cells (COMET) in humans in 2004, which represents the first use of a cultured non-limbal autologous cell type to treat bilateral LSCD. This review summarizes clinical outcomes from COMET studies published from 2004 to 2019 and reviews results with emphasis on the culture methods by which grafted cell sheets were prepared.

• Klíčová slova
• Cultivated oral mucosal epithelial cell, Limbal stem cell deficiency, Oral mucosal epithelial cells, Tissue regeneration,
• MeSH
• autologní transplantace MeSH
• epitelové buňky MeSH
• kultivované buňky MeSH
• lidé MeSH
• limbus corneae * MeSH
• nemoci rohovky * terapie   MeSH
• rohovkový epitel * MeSH
• transplantace buněk MeSH
• transplantace kmenových buněk MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

The probability of cell-to-cell coupling between directly adjacent cells (communication capability) in the V79-4 Chinese hamster cell line was evaluated under standard conditions or after 18-h treatment with EG. The cell monolayer did not form a continuous network of cells interconnected via gap junctions, but an average cell was coupled to only one half of its directly adjacent neighbours under standard conditions, or to one third of its directly neighbouring cells after 18-h exposure to EG. The rest of the directly adjacent neighbours did not establish functional gap junctions with an injected cell, although they were competent to couple to other cells with a probability similar to that of the coupling between the injected cell and its direct neighbours. Moreover, all the cells possessed the identical connexin--cx43, present on all cell membranes. The results indicated that the choice of a cell to which neighbour be coupled was rather random in the standard cell population as a whole, although the population contained some clones whose capability to couple was more or less different from that of the original cell population. Ethylene glycol reduced the gap junctional communication by increasing the frequency of cells not coupled to any of their direct neighbours from 1% for untreated cells to 23.3% for cells exposed to EG, and consequently by reducing the number of directly adjacent cells coupled to communicating injected cells. The communication capability of the cell population appeared to be unstable. It varied slightly in time and so did the response of the cells to EG. The results indicate that a cell can control its coupling to different directly adjacent neighbours independently, being able to control the gap junctional communication not only in time but in space as well. All control mechanisms of GJIC, known so far, affect a cell as a whole, while our results indicate that another regulatory mechanism may exist, controlling the gap junctional communication to different adjacent neighbouring cells independently.

• MeSH
• buněčná adheze účinky léků   MeSH
• časové faktory MeSH
• CHO buňky MeSH
• ethylenglykol farmakologie   MeSH
• fluorescenční barviva metabolismus   MeSH
• konexin 43 analýza   MeSH
• křečci praví MeSH
• mezerový spoj metabolismus   MeSH
• mezibuněčná komunikace * účinky léků   MeSH
• signální transdukce účinky léků   MeSH
• zvířata MeSH
• Check Tag
• křečci praví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ethylenglykol MeSH
• fluorescenční barviva MeSH
• konexin 43 MeSH

To replace the culture medium with a fresh one is a routine action of the mammalian cell culture technique. It is generally assumed that this act per se does not cause any significant physiological response of a cell population that would significantly interfere with the experimental procedures in culture. However, in this series of experiments we demonstrate that the exchange of the culture medium for a fresh one may induce a significant temporary decrease in the GJIC, assessed by the dye coupling method in the V79-4 Chinese hamster cell line. This effect is accompanied by a degradation of gap junctions and their re-establishment assessed by the semi-quantitative immunocytochemistry of connexin43. The minimum value of GJIC was reached 45 min after the exchange of the medium. Afterwards, GJIC grew up again, reaching the standard value 3 or 4 h later. This effect does not just result from the exchange of medium as a mechanical action, is not caused by the change of pH and is of quantitative character. The fresh medium loses its capability to reduce GJIC after 3 h of conditioning with the same cells. We found that the value of the early inhibition of GJIC observed during the first 2 h of treatment with the inhibitor of GJIC-EG (applied together with a fresh culture medium)--was indistinguishable from the effect of the exchange of medium itself. Only after that point of time is the EG-induced inhibition of GJIC definitely distinguishable. The results demonstrate that a simple exchange of the culture medium, which is generally implemented in various experiments in culture, may cause serious physiological, biochemical and even morphological responses of cells and thus affect the final results of experiments in culture, especially regarding the early effects of drugs. Consequently, to avoid an undesirable response of the cell population reported in this paper we recommend to apply or remove drugs using a medium conditioned with the same cells for at least 3 h.

• MeSH
• buněčné kultury metody   MeSH
• buněčné linie MeSH
• imunohistochemie MeSH
• isochinoliny MeSH
• konexin 43 metabolismus   MeSH
• křečci praví MeSH
• kultivační média speciální MeSH
• kultivační média MeSH
• mezerový spoj fyziologie   MeSH
• signální transdukce fyziologie   MeSH
• zvířata MeSH
• Check Tag
• křečci praví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• isochinoliny MeSH
• konexin 43 MeSH
• kultivační média speciální MeSH
• kultivační média MeSH
• lucifer yellow MeSH Prohlížeč

Neural rosette formation is a critical morphogenetic process during neural development, whereby neural stem cells are enclosed in rosette niches to equipoise proliferation and differentiation. How neural rosettes form and provide a regulatory micro-environment remains to be elucidated. We employed the human embryonic stem cell-based neural rosette system to investigate the structural development and function of neural rosettes. Our study shows that neural rosette formation consists of five types of morphological change: intercalation, constriction, polarization, elongation and lumen formation. Ca2+ signaling plays a pivotal role in the five steps by regulating the actions of the cytoskeletal complexes, actin, myosin II and tubulin during intercalation, constriction and elongation. These, in turn, control the polarizing elements, ZO-1, PARD3 and β-catenin during polarization and lumen production for neural rosette formation. We further demonstrate that the dismantlement of neural rosettes, mediated by the destruction of cytoskeletal elements, promotes neurogenesis and astrogenesis prematurely, indicating that an intact rosette structure is essential for orderly neural development.

• Klíčová slova
• Adherens junctions, Calcium, Cytoskeleton proteins, Embryonic stem cells, Neural rosette formation, Neural stem cell niche, Neurogenesis,
• MeSH
• aktiny metabolismus   MeSH
• apoptóza účinky léků   MeSH
• buněčný rodokmen účinky léků   MeSH
• cytoskelet účinky léků   metabolismus   MeSH
• lidé MeSH
• lidské embryonální kmenové buňky cytologie   účinky léků   metabolismus   MeSH
• myosin typu II metabolismus   MeSH
• nervové kmenové buňky cytologie   účinky léků   metabolismus   MeSH
• neurogeneze účinky léků   MeSH
• neurony cytologie   účinky léků   metabolismus   ultrastruktura   MeSH
• polarita buněk účinky léků   MeSH
• protein zonula occludens 1 metabolismus   MeSH
• tvar buňky * účinky léků   MeSH
• tvorba rozet * MeSH
• vápník farmakologie   MeSH
• vápníková signalizace * účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aktiny MeSH
• myosin typu II MeSH
• protein zonula occludens 1 MeSH
• vápník MeSH

Sensing of cytoplasmic DNA by cyclic guanosine monophosphate-adenosine monophosphate (cGAMP) synthase (cGAS) results in production of the dinucleotide cGAMP and consecutive activation of stimulator of interferon genes (STING) followed by production of type I interferon (IFN). Although cancer cells contain supra-normal concentrations of cytoplasmic DNA, they rarely produce type I IFN spontaneously. This suggests that defects in the DNA-sensing pathway may serve as an immune escape mechanism. We find that cancer cells produce cGAMP that is transferred via gap junctions to tumor-associated dendritic cells (DCs) and macrophages, which respond by producing type I IFN in situ. Cancer-cell-intrinsic expression of cGAS, but not STING, promotes infiltration by effector CD8+ T cells and consequently results in prolonged survival. Furthermore, cGAS-expressing cancers respond better to genotoxic treatments and immunotherapy. Thus, cancer-cell-derived cGAMP is crucial to protective anti-tumor CD8+ T cell immunity. Consequently, cancer-cell-intrinsic expression of cGAS determines tumor immunogenicity and makes tumors hot. These findings are relevant for genotoxic and immune therapies for cancer.

• Klíčová slova
• CD8(+) T cells, STING, cGAMP, cGAS, cancer, chemotherapy, gap junctions, immunotherapy, radiotherapy, type I IFN,
• MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• dendritické buňky metabolismus   MeSH
• imunoterapie MeSH
• interferon typ I metabolismus   MeSH
• lidé MeSH
• membránové proteiny MeSH
• mikrosatelitní repetice genetika   MeSH
• myši inbrední C57BL MeSH
• nádorové buněčné linie MeSH
• nádory farmakoterapie   imunologie   patologie   MeSH
• nukleotidy cyklické metabolismus   MeSH
• nukleotidyltransferasy metabolismus   MeSH
• poškození DNA MeSH
• progrese nemoci MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cGAS protein, human MeSH Prohlížeč
• cGAS protein, mouse MeSH Prohlížeč
• cyclic guanosine monophosphate-adenosine monophosphate MeSH Prohlížeč
• interferon typ I MeSH
• membránové proteiny MeSH
• nukleotidy cyklické MeSH
• nukleotidyltransferasy MeSH
• Sting1 protein, mouse MeSH Prohlížeč

Throughout the brain, astrocytes form networks mediated by gap junction channels that promote the activity of neuronal ensembles. Although their inputs on neuronal information processing are well established, how molecular gap junction channels shape neuronal network patterns remains unclear. Here, using astroglial connexin-deficient mice, in which astrocytes are disconnected and neuronal bursting patterns are abnormal, we show that astrocyte networks strengthen bursting activity via dynamic regulation of extracellular potassium levels, independently of glutamate homeostasis or metabolic support. Using a facilitation-depression model, we identify neuronal afterhyperpolarization as the key parameter underlying bursting pattern regulation by extracellular potassium in mice with disconnected astrocytes. We confirm this prediction experimentally and reveal that astroglial network control of extracellular potassium sustains neuronal afterhyperpolarization via KCNQ voltage-gated K+ channels. Altogether, these data delineate how astroglial gap junctions mechanistically strengthen neuronal population bursts and point to approaches for controlling aberrant activity in neurological diseases.

• Klíčová slova
• CP: Neuroscience, KCNQ channels, afterhyperpolarization, astrocytes, bursting, connexins, gap junctions, mathematical modeling, networks, potassium,
• MeSH
• akční potenciály fyziologie   MeSH
• astrocyty * metabolismus   MeSH
• draslík * metabolismus   MeSH
• draslíkové kanály KCNQ * metabolismus   genetika   MeSH
• hipokampus * metabolismus   MeSH
• konexiny metabolismus   genetika   MeSH
• mezerový spoj * metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši MeSH
• nervová síť metabolismus   MeSH
• neurony metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• draslík * MeSH
• draslíkové kanály KCNQ * MeSH
• konexiny MeSH

Calpain2 is a conventional member of the non-lysosomal calpain protease family that has been shown to affect the dynamics of focal and cell-cell adhesions by proteolyzing the components of adhesion complexes. Here, we inactivated calpain2 using CRISPR/Cas9 in epithelial MDCK cells. We show that depletion of calpain2 has multiple effects on cell morphology and function. Calpain2-depleted cells develop epithelial shape, however, they cover a smaller area, and cell clusters are more compact. Inactivation of calpain2 enhanced restoration of transepithelial electrical resistance after calcium switch, decreased cell migration, and delayed cell scattering induced by HGF/SF. In addition, calpain2 depletion prevented morphological changes induced by ERK2 overexpression. Interestingly, proteolysis of several calpain2 targets, including E-cadherin, β-catenin, talin, FAK, and paxillin, was not discernibly affected by calpain2 depletion. Taken together, these data suggest that calpain2 regulates the stability of cell-cell and cell-substratum adhesions indirectly without affecting the proteolysis of these adhesion complexes.

• Klíčová slova
• Actin, Adherens junctions, Calpains, Cell scattering, ERK, Epithelial polarity, Focal adhesions, HGF/SF, Migration, Proteases, Tight junctions, Transepithelial electrical resistance,
• MeSH
• beta-katenin metabolismus   MeSH
• buněčná adheze * MeSH
• buňky MDCK MeSH
• CRISPR-Cas systémy MeSH
• epitelové buňky * metabolismus   cytologie   MeSH
• hepatocytární růstový faktor metabolismus   MeSH
• kadheriny metabolismus   MeSH
• kalpain * metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 1 metabolismus   MeSH
• pohyb buněk MeSH
• proteolýza MeSH
• psi MeSH
• vápník metabolismus   MeSH
• zvířata MeSH
• Check Tag
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• beta-katenin MeSH
• hepatocytární růstový faktor MeSH
• kadheriny MeSH
• kalpain * MeSH
• mitogenem aktivovaná proteinkinasa 1 MeSH
• vápník MeSH

VEGFR-2/Notch signalling regulates angiogenesis in part by driving the remodelling of endothelial cell junctions and by inducing cell migration. Here, we show that VEGF-induced polarized cell elongation increases cell perimeter and decreases the relative VE-cadherin concentration at junctions, triggering polarized formation of actin-driven junction-associated intermittent lamellipodia (JAIL) under control of the WASP/WAVE/ARP2/3 complex. JAIL allow formation of new VE-cadherin adhesion sites that are critical for cell migration and monolayer integrity. Whereas at the leading edge of the cell, large JAIL drive cell migration with supportive contraction, lateral junctions show small JAIL that allow relative cell movement. VEGFR-2 activation initiates cell elongation through dephosphorylation of junctional myosin light chain II, which leads to a local loss of tension to induce JAIL-mediated junctional remodelling. These events require both microtubules and polarized Rac activity. Together, we propose a model where polarized JAIL formation drives directed cell migration and junctional remodelling during sprouting angiogenesis.

• MeSH
• aktiny účinky léků   metabolismus   MeSH
• buněčná adheze MeSH
• CD antigeny účinky léků   metabolismus   MeSH
• cévní endotel MeSH
• endoteliální buňky pupečníkové žíly (lidské) MeSH
• endoteliální buňky účinky léků   metabolismus   fyziologie   MeSH
• fyziologická neovaskularizace účinky léků   fyziologie   MeSH
• kadheriny účinky léků   metabolismus   MeSH
• komplex proteinů 2-3 souvisejících s aktinem metabolismus   MeSH
• lehké řetězce myosinu metabolismus   MeSH
• lidé MeSH
• mezibuněčné spoje účinky léků   metabolismus   MeSH
• mikrotubuly účinky léků   metabolismus   MeSH
• modely kardiovaskulární MeSH
• pohyb buněk účinky léků   fyziologie   MeSH
• polarita buněk účinky léků   fyziologie   MeSH
• protein 2 související s aktinem metabolismus   MeSH
• protein 3 související s aktinem metabolismus   MeSH
• protein Wiskottova-Aldrichova syndromu metabolismus   MeSH
• pseudopodia účinky léků   metabolismus   fyziologie   MeSH
• rac proteiny vázající GTP metabolismus   MeSH
• receptor 2 pro vaskulární endoteliální růstový faktor metabolismus   MeSH
• remodelace cév MeSH
• rodina proteinů Wiskottova-Aldrichova syndromu metabolismus   MeSH
• signální transdukce MeSH
• srdeční myosiny metabolismus   MeSH
• vaskulární endoteliální růstový faktor A metabolismus   farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ACTR2 protein, human MeSH Prohlížeč
• ACTR3 protein, human MeSH Prohlížeč
• aktiny MeSH
• cadherin 5 MeSH Prohlížeč
• CD antigeny MeSH
• kadheriny MeSH
• KDR protein, human MeSH Prohlížeč
• komplex proteinů 2-3 souvisejících s aktinem MeSH
• lehké řetězce myosinu MeSH
• myosin light chain 2 MeSH Prohlížeč
• protein 2 související s aktinem MeSH
• protein 3 související s aktinem MeSH
• protein Wiskottova-Aldrichova syndromu MeSH
• rac proteiny vázající GTP MeSH
• receptor 2 pro vaskulární endoteliální růstový faktor MeSH
• rodina proteinů Wiskottova-Aldrichova syndromu MeSH
• srdeční myosiny MeSH
• vaskulární endoteliální růstový faktor A MeSH
• VEGFA protein, human MeSH Prohlížeč
• WAS protein, human MeSH Prohlížeč
• WASF1 protein, human MeSH Prohlížeč