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1.
Summary The intercellular contacts of the migrating edge of chick and quail blastoderms during gastrulation were studied by transmission electron microscopy of thin sections and of freeze-fracture replicas. Tight junctions and gap junctions as well as desmosomes were found. Tight junctions were organized as single junctional strands or as a complex of numerous junctional strands interposed between the lamellae and the bodies of the cells building up the margin of overgrowth. The function of these intercellular junctions is considered in relation to the locomotion of the margin of overgrowth cells. 相似文献
2.
MDCK cell monolayers grown on glass coverslips were used to examine the Na+ concentration in individual lateral intercellular spaces (LIS) by video fluorescence microscopy. The LIS was filled with the Na+-sensitive fluorescent dye SBFO by incubation of the monolayers for 75–90 min with 250 m of the membrane impermeant form of the dye. After dye loading, the monolayers were perfused at 37°C with solutions buffered with HEPES or bicarbonate/CO2 containing 142 mm Na+. Ratios of the fluorescence images after sequential excitation with 340 nm and 380 nm light were performed and in situ calibration of LIS Na+ was accomplished after blocking the Na+ pump with 5 × 10–4 m ouabain. Measurements of Na+ along the basolateral-to-apical axis of the LIS at 1.0 or 1.5 m intervals did not reveal a Na+ gradient when the perfusate was either HEPES or bicarbonate/CO2 solutions. In bicarbonate solutions, the mean Na+ concentration (mm) was 157.2 ± 2.3, 15 mm higher than the bath Na+ concentration. In HEPES solutions, however, the Na+ concentration was not different from the bath concentration (142.7 ± 3.1 mm). The time course of Na+ changes in LIS was investigated by rapidly switching the perfusate from 142 to 80 mm Na+ and measuring the Na+ changes at one focal plane.We would like to thank P.H. Tran and C. Gibson for their technical and computational assistance as well as Dr. B.-E. Persson (University of Uppsala, Sweden) for his contribution in the early phases of the study. 相似文献
3.
Prostaglandins are important in signaling pathways involved in modulating the rates of Na+ transport in a diverse group of tissues possessing apical membrane epithelial channels. PGE2 is known to cause either stimulation, inhibition or transient stimulatory changes of Na+ transport. We have continued our studies of frog skins that are known to respond to forskolin and PGE2 with large steady-state increases of transport and have used noninvasive methods of blocker-induced noise analysis of Na+ channels to determine their channel densities (N
T
) and open probabilities (P
o
). In the absence of exogenous hormones, baseline rates of Na+ transport are especially high in scraped skins (R. pipiens pipiens) studied in the fall of the year. Na+ transport was inhibited by indomethacin and by removal of the unstirred layers of the corium (isolated epithelia) alone suggesting
that PGE2 is responsible for the sustained and elevated rates of transport in scraped skins. Changes of transport caused by indomethacin,
forskolin or PGE2 were unquestionably mediated by considerably larger changes of N
T
than compensatory changes of P
o
. Since cAMP caused no change of P
o
in tissues pretreated with indomethacin, PGE2 appears in this tissue to serve a dual role, increasing the steady state N
T
by way of cAMP and decreasing P
o
by unknown mechanisms. Despite appreciable PGE2-related decreases of P
o
, the net stimulation of transport occurs by a considerably greater cAMP-mediated increase of N
T
.
Received: 28 February 1996/Revised: 22 August 1996 相似文献
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David E. Saslowsky Yvonne M. te Welscher Daniel J.-F. Chinnapen Jessica S. Wagner Joy Wan Eli Kern Wayne I. Lencer 《The Journal of biological chemistry》2013,288(36):25804-25809
Cholera toxin causes diarrheal disease by binding ganglioside GM1 on the apical membrane of polarized intestinal epithelial cells and trafficking retrograde through sorting endosomes, the trans-Golgi network (TGN), and into the endoplasmic reticulum. A fraction of toxin also moves from endosomes across the cell to the basolateral plasma membrane by transcytosis, thus breeching the intestinal barrier. Here we find that sorting of cholera toxin into this transcytotic pathway bypasses retrograde transport to the TGN. We also find that GM1 sphingolipids can traffic from apical to basolateral membranes by transcytosis in the absence of toxin binding but only if the GM1 species contain cis-unsaturated or short acyl chains in the ceramide domain. We found previously that the same GM1 species are needed to efficiently traffic retrograde into the TGN and endoplasmic reticulum and into the recycling endosome, implicating a shared mechanism of action for sorting by lipid shape among these pathways. 相似文献
6.
Separation of distinct body, organ and tissue compartments, and maintenance of epithelial cell polarity require tight junctions (TJ)--cell-cell junctions located in the apicolateral regions of epithelial and endothelial cells. Studies on the protein components of vertebrate TJ have revealed an intricate network of membrane, sub-membrane, cytoskeletal, and signalling molecules. How these molecules functionally interact to provide TJ with their functions, and what roles these molecules play in control of cell growth and differentiation is a fundamental problem in cell biology. 相似文献
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Diseases of the gastrointestinal system are often related with irritations or pathological changes of mucous membranes. In an ex vivo system based on porcine colonic tissue various neutral and acidic polysaccharides were tested concerning their bioadhesive potential in order to form artificial mucin layers on colon epithelial membranes. Rhamnogalacturonans with a low degree of esterification and linear oligogalacturonids derived from pectin showed significant bioadhesion against colonic mucous membranes. In contrast highly esterified pectins and neutral polysaccharides were ineffective. Within a structure–activity relationship linear, strongly acidic homogalacturonides were shown to be most adhesive agents. Esterification, branching or non-linear backbone structures will reduce the adhesive properties. The bioadhesive effects were concentration-dependent. Polysaccharide layers, located exclusively on the apical membrane surface of colonic tissue, were visualized by fluorescent microscopy. The adhesion of the exogenous galacturonides on the tissue surface was mediated by interaction with the endogenous mucin, for the release of the endogenous mucines with a mucolytic agent resulted in a decreased bioadhesion of exogenous galacturonides. Additionally, mucin–galacturonide synergism was shown by rheological methods. The artificial mucin layers provide protective effects on colonic mucous membranes against toxic agents as shown by incubation of the tissue with TritonX-100. 相似文献
9.
V. Calderón A. Lázaro R.G. Contreras L. Shoshani C. Flores-Maldonado L. González-Mariscal G. Zampighi M. Cereijido 《The Journal of membrane biology》1998,164(1):59-69
Tight junctions (TJs) are cell-to-cell contacts made of strands, which appear as ridges on P faces and complementary furrows
on E faces on freeze fracture replicas. Evidences and opinions on whether these strands are composed of either membrane-bound
proteins or lipid micelles are somewhat varied. In the present work we alter the lipid composition of Madin-Darby canine kidney
monolayers using a novel approach, while studying (i) their transepithelial electrical resistance, a parameter that depends
on the degree of sealing of the TJs; (ii) the apical-to-basolateral flux of 4 kD fluorescent dextran (JDEX), that reflects the permeability of the intercellular spaces; (iii) the ability of TJs to restrict apical-to-basolateral
diffusion of membrane lipids; and (iv) the pattern of distribution of endogenous and transfected occludin, the sole membrane
protein presently known to form part of the TJs. We show that changing the total composition of phospholipids, sphingolipids,
cholesterol and the content of fatty acids, does not alter TER nor the structure of the strands. Interestingly, enrichment
with linoleic acid increases the JDEX by 631%. The fact that this increase is not reflected in a decrease of TER, suggests that junctional strands do not act as
simple resistive elements but may contain mobile translocating mechanisms.
Received: 7 November 1997/Revised: 20 March 1998 相似文献
10.
Phillip L. Butler Alexander Staruschenko Peter M. Snyder 《The Journal of biological chemistry》2015,290(20):12497-12503
The epithelial Na+ channel (ENaC) functions as a pathway for Na+ absorption in the kidney and lung, where it is crucial for Na+ homeostasis and blood pressure regulation. ENaC is regulated in part through signaling pathways that control the ubiquitination state of ENaC lysines. A defect in ubiquitination causes Liddle syndrome, an inherited form of hypertension. Here we determined that α-, β-, and γENaC are also substrates for lysine acetylation. Trichostatin A (TSA), a histone deacetylase inhibitor, enhanced ENaC acetylation and increased ENaC abundance in the total cell lysate and at the cell surface. Moreover, TSA increased ENaC current in Fischer rat thyroid and kidney collecting duct epithelia. We found that HDAC7 is expressed in the kidney collecting duct, supporting a potential role for this histone deacetylase in ENaC regulation. HDAC7 overexpression reduced ENaC abundance and ENaC current, whereas ENaC abundance and current were increased by silencing of HDAC7. ENaC and HDAC7 form a complex, as detected by coimmunoprecipitation. We observed a reciprocal relationship between acetylation and ubiquitination; TSA reduced ENaC ubiquitination, whereas HDAC7 increased ubiquitination. By reducing ENaC ubiquitination, TSA decreased the rate of ENaC degradation. Thus, acetylation increases epithelial Na+ absorption by antagonizing ENaC ubiquitination. This stabilizes ENaC, and hence, increases its abundance at the cell surface. 相似文献