Immunolocalization of protease-activated receptors in endothelial cells of splenic sinuses.

The immunolocalization of protease-activated receptors (PARs) and related proteins in splenic sinus endothelial cells was examined using immunofluorescence and electron microscopy. Immunofluorescence microscopy showed that PAR1 colocalized with PAR2, PAR3, and PAR4. PAR4 colocalized with PAR3 and P2Y12.

Immunohistochemical study of dissociation and association of adherens junctions in splenic sinus endothelial cells.

It is not yet clear whether cellular junctions between splenic sinus endothelial cells are open or closed. In order to clarify this, immunolocalization of thrombomodulin (TM), endothelial protein C receptor (EPCR), protease-activated receptor 1 (PAR1), sphingosine 1-phosphate receptor 1 (S1P), β-catenin phosphorylated at Try142 (β-catenin Y142) and β-catenin phosphorylated at Try654 (β-catenin Y654), which are related proteins that regulate dissociation and association of the adherens junctions of endothelial cells, are examined in rats using laser microscopy and electron microscopy. TM, EPCR, PAR1 and S1P were colocalized in the entire circumference of the endothelial cells, as well as in the caveolar membranes and junctional membranes of adjacent endothelial cells.

Extensive Ca2+ leak through K4750Q cardiac ryanodine receptors caused by cytosolic and luminal Ca2+ hypersensitivity.

Various ryanodine receptor 2 (RyR2) point mutations cause catecholamine-induced polymorphic ventricular tachycardia (CPVT), a life-threatening arrhythmia evoked by diastolic intracellular Ca release dysfunction. These mutations occur in essential regions of RyR2 that regulate Ca release. The molecular dysfunction caused by CPVT-associated RyR2 mutations as well as the functional consequences remain unresolved.

Differentiated localizations of phosphorylated focal adhesion kinase in endothelial cells of rat splenic sinus.

The splenic sinus endothelium adhering via adherens junctions and tight junctions regulates the passage of blood cells through the splenic cord. Focal adhesion kinase (FAK) regulates the focal adhesion complex in the basal part of endothelial cells and is an integrated component of cell-cell adhesion, depending on its phosphorylation status. The objectives of this study are to assess the localization of FAK phosphorylated at tyrosine residues and the related proteins of integrin β5, talin, paxillin, p130Cas, vinculin, RhoA, Rac1, Rac2, Cdc42 and VE-cadherin, in the sinus endothelial cells of rat spleen and to examine the roles of FAK in regulating endothelial adhesion and the passage of blood cells.

Integrin αvβ5 in endothelial cells of rat splenic sinus: an immunohistochemical and ultrastructural study.

Localization of integrins β1-8, α1, α2, α3, α5, α6 and αv in sinus endothelial cells of the rat spleen was examined by immunofluorescence microscopy. Labeling for anti-integrin β5 and integrin αv was detected and colocalized in the entire circumference of endothelial cells. Labeling for integrin β5, vinculin and actin filaments demonstrated that they lay close to each other in the basal part of the endothelial cells.

CLCA splicing isoform associated with adhesion through β1-integrin and its scaffolding protein: specific expression in undifferentiated epithelial cells.

We previously found that a rat CLCA homologue (rCLCA-f) modulates Ca(2+)-dependent Cl(-) transport in the ductal cells of the rat submandibular gland. CLCA proteins have been shown to be multifunctional, with roles in, for example, cell adhesion. Here, we describe the mRNA and protein expressions of a splicing isoform of rat rCLCA (rCLCA-t).

P2Y1, P2Y6, and P2Y12 receptors in rat splenic sinus endothelial cells: an immunohistochemical and ultrastructural study.

Localization of three P2X and six P2Y receptors in sinus endothelial cells of the rat spleen was examined by immunofluorescent microscopy, and ultrastructural localization of the detected receptors was examined by immunogold electron microscopy. In immunofluorescent microscopy, labeling for anti-P2Y1, P2Y6, and P2Y12 receptors was detected in endothelial cells, but P2X1, P2X2, P2X4, P2Y2, P2Y4, and P2Y13 receptors was not detected. P2Y1 and P2Y12 receptors were prominently localized in the basal parts of endothelial cells.

Vimentin intermediate filaments: the central base in sinus endothelial cells of the rat spleen.

The ultrastructural distribution of vimentin intermediate filaments (IFs) and localizations of the related proteins in sinus endothelial cells of the rat spleen was examined by confocal laser scanning and electron microscopy with detergent extraction, myosin-fragment 1 decoration, and immunogold labeling to elucidate their functions in endothelial cells. Vimentin IFs were extremely abundant over stress fibers in the basal part of the endothelial cells. Some of them were intermingled with actin filaments in stress fibers, and were associated with coated vesicles.

Altered KCNQ3 potassium channel function caused by the W309R pore-helix mutation found in human epilepsy.

The second tryptophan (W) residue of the conserved WW motif in the pore helix of many K+ channel subunit is thought to interact with the tyrosine (Y) residues of the selectivity filter. A missense mutation causing the replacement of the corresponding residues with an arginine (W309R) occurs in KCNQ3 subunits forming part of M-channels. In this study, we examined the functional consequences of the W309R mutation in heterogously expressed KCNQ channels.

Localization of claudin-5 and ZO-1 in rat spleen sinus endothelial cells.

Splenic sinus endothelial cells, which adhere through tight and adherens junctions, regulate the passage of blood cells through the splenic cord. The objective of this study was to assess the localization of tight junctional proteins, claudin-5 and ZO-1 in the sinus endothelial cells of rat spleen and to characterize spatial and functional relationships between tight and adherens junctions. Immunofluorescence microscopy of tissue cryosections demonstrated that claudin-5, ZO-1, and alpha-catenin were distinctly localized in the junctional regions of adjacent endothelial cells.

Cell membrane-derived lysophosphatidylcholine activates cardiac ryanodine receptor channels.

Lysophosphatidylcholine (LPC) is metabolized from a membrane phospholipid and modulates a variety of channels in the plasma membrane (PM). We examined LPC modulation of cardiac ryanodine receptor (RyR) channels in the sarcoplasmic reticulum (SR) using the planar lipid bilayer method to measure the single-channel currents. Micromolar concentrations of LPC increased the open probability of the reconstituted RyR channels irrespective of whether LPC was added to the cis or trans chamber.

Distribution of adherens junction mediated by VE-cadherin complex in rat spleen sinus endothelial cells.

The splenic sinus endothelium regulates the passage of blood cells through the splenic cord. The goal of the present study was to assess the localization of vascular endothelial (VE)-cadherin, beta-catenin, and p120-catenin in the sinus endothelial cells of rat spleen and to characterize the presence and distribution of adherens junction formation mediated by the cadherin-catenin complex. Immunofluorescent microscopy of tissue cryosections demonstrated that VE-cadherin, beta-catenin, and p120-catenin were localized in the junctional regions of adjacent endothelial cells.

Localization of TRPC1 channel in the sinus endothelial cells of rat spleen.

The ultrastructural localization of transient receptor potential C1 (TRPC1) channels in the sinus endothelial cells of rat spleen was examined by confocal laser scanning and electron microscopy. In addition, the localization of the closely associated proteins and channels, VE-cadherin, calreticulin, inositol-1,4,5-trisphosphate receptors type 1 (IP3R1), and ryanodine receptor (RyR), was also examined. Immunofluorescence microscopy of tissue cryosections revealed TRPC1 channels to be localized within the cytoplasm, in the superficial layer of the apical and basal parts of the cells, and in the junctional area of the adjacent endothelial cells.

Localization of ryanodine receptor 3 in the sinus endothelial cells of the rat spleen.

The ultrastructural localization of ryanodine receptors (RyR) in sinus endothelial cells of the rat spleen was examined by confocal laser scanning and electron microscopy by using isoform-specific antibodies to each of the RyR isoforms. Immunofluorescence microscopy of tissue cryosections revealed RyR3 to be localized, with a strand-like form, in the superficial layer and within the cytoplasm of endothelial cells. Antibodies to RyR1 and RyR2 did not react indicating RyR3 was the predominant isoform.

Localization of caveolin-3 in the sinus endothelial cells of the rat spleen.

The localization of caveolins in the sinus endothelial cells of the rat spleen has been demonstrated by confocal laser scanning and electron microscopy. Caveolin-3, a muscle-specific caveolin, was detected by Western blot analysis and immunofluorescence microscopy of isolated sinus endothelial cells and tissue cryosections of the spleen. During the immunofluorescence microscopy of isolated endothelial cells, both caveolin-3 and caveolin-1 were found.

Structure of comblike teeth of the ayu sweetfish, Plecoglossus altivelis (Teleostei: Isospondyli): I. Denticles and tooth attachment.

The structure and tooth attachment of the comblike teeth and denticles of the ayu sweetfish, Plecoglossus altivelis, were examined by light and scanning electron microscopy. The denticle is composed of a spoonlike crown with a spine pointed anteriorly, a triangular plate in the cervical region, and a root that curves laterally and tapers off to a point. The root apex is fused with a long thin pedicle that turns abruptly anteriad toward the jaw bone.