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L Landmann

Publications and source records attributed to L Landmann.

At least 37 records · Page 2Linked to original sources
PubMed1997

Neural agrin induces ectopic postsynaptic specializations in innervated muscle fibers.

Neural agrin, in the absence of a nerve terminal, can induce the activity-resistant expression of acetylcholine receptor (AChR) subunit genes and the clustering of synapse-specific adult-type AChR channels in nonsynaptic regions of adult skeletal muscle fibers. Here we show that, when expression plasmids for neural agrin are injected into the extrasynaptic region of innervated muscle fibers, the following components of the postsynaptic apparatus are aggregated and colocalized with ectopic agrin-induced AChR clusters: laminin-beta2, MuSK, phosphotyrosine-containing proteins, beta-dystroglycan, utrophin, and rapsyn. These components have been implicated to play a role in the differentiation of neuromuscular junctions. Furthermore, ErbB2 and ErbB3, which are thought to be involved in the regulation of neurally induced AChR subunit gene expression, were colocalized with agrin-induced AChR aggregates at ectopic nerve-free sites. The postsynaptic muscle membrane also contained a high concentration of voltage-gated Na+ channels as well as deep, basal lamina-containing invaginations comparable to the secondary synaptic folds of normal endplates. The ability to induce AChR aggregation in vivo was not observed in experiments with a muscle-specific agrin isoform. Thus, a motor neuron-specific agrin isoform is sufficient to induce a full ectopic postsynaptic apparatus in muscle fibers kept electrically active at their original endplate sites.

Agrin↗
PubMed1996

Bovine endothelial-like cells increase intercellular junctions under treatment with interferon-gamma. An in vitro study.

We have recently shown that a subgroup of endothelial-like cells from the bovine corpus luteum maintains cytokeratin (CK) expression in long-term culture. Treatment with interferon-gamma (IFN-gamma) for three days brought about an increase in the intercellular junctions in CK-positive postconfluent cultures, as shown by immunofluorescence with specific monoclonal antibodies and by electron microscopy. Desmosomes, zonulae adhaerentes and both tight and gap junctions were found. The intercellular junctions (excluding the gap junctions) had increased in amount, while permeability of the cell layer to horseradish peroxidase (HRP) had decreased. Hence, IFN-gamma has reinforced intercellular junctions in our CK-positive endothelial-like cells.

Animals↗
PubMed1996

Structure-function correlation of tight junctional impairment after intrahepatic and extrahepatic cholestasis in rat liver.

BACKGROUND & AIMS: Tight junctions, the only barrier between blood and bile, are crucial in bile formation. The aim of this study was to correlate changes in morphology and permeability by comparing structural parameters with marker secretion into normal and cholestatic rat bile. METHODS: Cholestasis was induced by bile duct ligation of 5 and 21 days of ethinylestradiol administration. Quantitated structural parameters induced junctional length, strand number, junctional depth, and spacing of junctional particles. Junctional permeability was probed with horseradish peroxidase and dextrans of increasing sizes. RESULTS: Junctional length was decreased slightly by ethinylestradiol (-16% after 21 days) but increased by ligation (77%). Mean strand number decreased from 4.6 to 3.7 after 21 days of ethinylestradiol and 3.4 after ligation associated with increased junctional depth. The proportions of morphologically horseradish peroxidase-positive junctions increased from 4% to 15% after 21 days of ethinylestradiol and to 56% after ligation. Horseradish peroxidase secretion was increased twofold by ethinylestradiol and 6.5-fold by ligation, paralleled by an increase of dextran size selectivity from 70,000 to 79,000 daltons after ethinylestradiol and to 266,000 daltons after ligation. CONCLUSIONS: Impairment of junctional integrity is paralleled with the degree of cholestasis, whereas correlation of morphological and physiological alterations shows a close structure-function relationship.

Animals↗
PubMed1995

Cholestasis-induced alterations of the trans- and paracellular pathways in rat hepatocytes.

Bile secretion depends on the vectorial transport of solutes from blood to bile and involves three different pathways: transcellular pathways mediated by transport proteins distributed asymmetrically in the basolateral and canalicular plasma membrane and by transcytotic vesicles, and a paracellular pathway allowing selective diffusion through tight junctions. All three pathways are impaired differentially by extrahepatic (bile duct ligation) or intrahepatic (ethinyloestradiol) cholestasis. Ethinyloestradiol treatment leads to tight junctional defects that are less severe than those induced by bile duct ligation. Junctional impairment is reflected functionally in increased permeability for horseradish peroxidase and structurally by decreased strand numbers and increased junctional length, but not by alterations at the level of the individual strands. The parallelism of physiological and morphological perturbations indicates a structure-function relationship in hepatocellular tight junctions. In addition, impaired functional integrity of tight junctions following bile duct ligation is reflected in a partial loss of hepatocellular surface polarity owing to redistribution of some, but not all, domain-specific plasma membrane antigens, which might mimic the behaviour of transport systems. After ethinyloestradiol treatment no alterations of surface polarity were observed. Thus, immunohistochemistry supports the view that ethinyloestradiol results in less severe impairment of the tight junctions than bile duct ligation. Finally, bile duct ligation, but not ethinyloestradiol, affects the transcytotic vesicular pathway; severe impairment of this is reflected in the absence of a late horseradish peroxidase peak in bile and also in the accumulation of pericanalicular vesicles that are immunopositive for canalicular membrane proteins and accessible for bulk phase endocytic markers.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

In situ localization of the hepatocytic Na+/Taurocholate cotransporting polypeptide in rat liver.

BACKGROUND/AIMS: An Na+/taurocholate cotransporting polypeptide (Ntcp) has recently been cloned from rat liver. The aim of this study was to directly characterize the native Ntcp on the protein level and study its in situ distribution in rat liver. METHODS: A rabbit antiserum was raised against a fusion protein containing the maltose-binding protein and the C terminus of Ntcp. Native Ntcp was localized in situ by immunofluorescent techniques. Expression of Ntcp was directly correlated with taurocholate uptake measurements in stably transfected Chinese hamster ovary cells. RESULTS: Native Ntcp showed an apparent molecular weight of 51,000. After deglycosylation of isolated basolateral rat liver plasma membranes, the apparent molecular weight of Ntcp decreased to 33,500. In intact rat liver, Ntcp was selectively localized at the basolateral surface domain of hepatocytes. In short-term cultured hepatocytes, a positive surface immunoreaction was only obtained in detergent-permeabilized cell cultures. In stably transfected Chinese hamster ovary cells, the surface expression of immunopositive Ntcp was associated with Na(+)-dependent taurocholate uptake activity. CONCLUSIONS: Native Ntcp represents a glycoprotein of the basolateral hepatocyte plasma membrane with its C-terminal end facing the intracellular compartment. Furthermore, surface expression of Ntcp is a prerequisite for Na(+)-dependent taurocholate uptake to occur, thus providing further proof for its bile acid transport function in rat liver.

Animals↗
PubMed1994

Effect of obstructive cholestasis on membrane traffic and domain-specific expression of plasma membrane proteins in rat liver parenchymal cells.

We investigated the effect of bile duct ligation and its release on membrane traffic and plasma membrane protein distribution in rat hepatocytes. Immunofluorescence studies with monoclonal antibodies against six domain-specific surface antigens revealed that bile duct ligation leads to an accumulation of pericanalicular vesicles containing canalicular antigens. All apical antigens could be demonstrated in the basolateral plasma membrane, whereas only one out of three basolateral antigens redistributed to the canalicular plasma membrane. After release of bile duct ligation, the accumulated pericanalicular vesicles disappeared within minutes, whereas the plasma membrane polarity was not restored within 1 hr. Monitoring secretion of polymeric IgA and horseradish peroxidase into bile demonstrated that bile duct ligation also inhibits the transcytotic vesicle pathway and severely impairs the function of tight junctions. In contrast, bile duct ligation appears not to affect the endoplasmic reticulum to basolateral membrane traffic as assessed by determination of newly synthesized albumin and transferrin in serum nor does it influence receptor mediated endocytosis at the basolateral plasma membrane.

Animals↗
PubMed1992

Ethinylestradiol increases volume and decreases sinusoidal membrane surface in the rat liver: a stereological analysis.

Structural alterations of liver parenchyma caused by ethinylestradiol, a synthetic estrogen known to induce cholestasis and to act as a tumor promoter factor, were investigated. Male rats treated with 17 alpha-ethinylestradiol (5 mg/kg body weight for 5 days) were compared with controls (n = 5 each). After perfusion fixation and systematic random sampling, paraffin sections, semithin sections and thin sections were examined observing standard stereological techniques. Ethinylestradiol treatment induced an increase in liver volume by 65% (p less than 0.001), which was caused more by hypertrophy (volume of singular hepatocyte +35%, p less than 0.001) than by hyperplasia (number of hepatocytes +23%, p less than 0.001). A decrease in sinusoidal membrane surface density (-43%, p less than 0.005) associated with a decrease in sinusoidal microvillar volume density (-50%, p less than 0.005) were both compensated for by the increase in liver volume. No canalicular alterations were observed. Thus changes in hepatocytes detectable with stereological techniques affect the sinusoidal pole where decreased sinusoidal membrane surface is associated with or reflects a substantial loss of membrane phospholipids. The increased liver volume may constitute an adaptive response compensating for the relative decrease in sinusoidal membrane surface and displays characteristics comparable to those of preneoplastic hepatocytes.

Animals↗
PubMed1990

Bile duct ligation-induced redistribution of canalicular antigen in rat hepatocyte plasma membranes demonstrated by immunogold quantitation.

Extrahepatic obstructive cholestasis has been demonstrated to induce a redistribution of domain specific membrane proteins in rat hepatocytes reflecting loss or even reversal of cell polarity. In order to further characterize the redistribution of canalicular antigens, we used the Lowicryl K4M immunogold technique for examination of the effects of bile duct ligation (50 h) on the distribution of antigen in rat hepatocytes at the ultrastructural level and quantitated immuno-gold density in the three domains of the plasma membrane. In normal hepatocytes, antigen was localized almost exclusively in the canalicular domain while the sinusoidal and lateral membranes showed only weak immunoreactivity. Other localizations included organelles compatible with known pathways of biosynthesis and degradation. Bile duct ligation markedly reduced immunolabel in the canalicular and increased it slightly in the sinusoidal domain. The number and staining intensity of immunoreactive subcanalicular lysosomes and vesicles probably representing endosomes was augmented. Number of immunogold particles per micron of plasma membrane were 7.86 vs 2.46 (P less than 0.005) in the canalicular, 1.16 vs 1.38 (n.s.) in the sinusoidal, and 1.23 vs 1.08 (n.s.) in the lateral domain resulting in a canalicular decrease by 68.7% and a sinusoidal increase of 19.0%. Overall decrease in total plasma membranes was by 29.7% (P less than 0.05). Thus, our data show that the sinusoidal and lateral domains behave differently. Furthermore, quantitative immunocytochemistry demonstrates a decrease in the canalicular antigen density and suggests a sinusoidal increase. The present data agree with the concept that bile duct ligation results in a loss or even reversal of cell polarity in hepatocytes.

Animals↗
PubMed1990

Decreased hepatocellular volume and intact morphology of tight junctions in calcium deprivation-induced cholestasis. Stereological and multiple indicator dilution analysis.

Cholestasis induced by perfusion of the liver with hypocalcemic media has been ascribed to several defects in bile secretion including increased biliary permeability. To investigate this model of cholestasis further, livers perfused with hypo- and normocalcemic media were examined stereologically using thin sections and freeze-fracture replicas. Organization of tight junctions was not altered by hypocalcemia; neither the number of strands nor the junctional depth were significantly affected. By contrast, the volume of hepatocytes decreased by 11% (p less than 0.001), compensated for by an increase in the space of Dissé and of the sinusoids. The canalicular length decreased by 25% (p less than 0.01), while the canalicular membrane surface was not altered. Multiple indicator dilution studies confirmed a decrease in hepatocellular volume, measured as the water space by 14% (p less than 0.03). This was compensated for by an increase in the extravascular sucrose, but not the albumin space. Immediately after switching from normo- to hypocalcemic perfusate a K+ efflux of 62 mumol/g liver was observed corresponding to approx. 8% of the hepatocellular water space. Our results suggest that hypocalcemia-induced cholestasis is due, at least in part, to a disturbance of the osmotic equilibrium, possibly caused by impairment of an ion transport system involved in hepatocellular volume control.

Animals↗
PubMed1989

Does a detailed explanation prior to gastroscopy reduce the patient's anxiety?

The object of this study was to investigate the assumption that a detailed explanation and description of gastroscopy to a patient reduces his fears of the procedure. In order to check this assumption, 243 consecutive patients who underwent elective gastroscopy were randomly distributed to 4 groups: Group A received a standard brief description of upper G-I endoscopy by the treating physician; Group B patients were given a detailed description by the endoscopist himself; Group C was given a comprehensive explanation and was shown an album of pictures illustrating each stage of the procedure; Group D saw a specially prepared video film of the procedure. A fifth group, E, consisted of patients who had previously undergone upper gastrointestinal endoscopy. Patient anxiety was measured using the "Spielberger State and Trait Anxiety Scales". According to the scoring on these scales there was no significant difference between the groups, thus showing that increasingly detailed description of the procedure did not diminish patients' fear. Neither did ethnic background or age have any significant influence on these results. Higher education had marginal influence on patients' apprehension about the procedure. However, female patients had significantly higher anxiety state and trait anxiety scoring than men. If endoscopy can be taken as typically representative of all invasive procedures, these results may have a much more general application.

Adolescent↗
PubMed1989

Extrahepatic obstructive cholestasis reverses the bile salt secretory polarity of rat hepatocytes.

To elucidate the consequences of extrahepatic cholestasis on the structure and function of hepatocytes, we studied the effects of bile duct ligation on the turnover, surface distribution, and functional activity of the canalicular 100-kD bile salt transport protein (cBSTP). Basolateral (blLPM) and canalicular (cLPM) liver plasma membrane vesicles were purified to the same degree from normal and cholestatic rat livers and the membrane bound cBSTP identified and quantitated using polyclonal anti-cBSTP antibodies. Cholestasis of 50 h resulted in an increased release of cBSTP into bile, thereby decreasing its in vivo half-life from 65 to 25 h. Furthermore, a significant portion of cBSTP accumulated at the basolateral surface and in intracellular vesicles of cholestatic hepatocytes. This redistribution of cBSTP was functionally paralleled by decreased and increased electrogenic taurocholate anion transport in cLPM and blLPM vesicles, respectively. These results demonstrate that biliary obstruction causes a reversal of the bile salt secretory polarity of rat hepatocytes. The resulting increase in basolateral (sinusoidal) bile salt efflux might protect hepatocytes from too high an accumulation of toxic bile salts within the cell interior.

Animals↗
PubMed1988

The epidermal permeability barrier.

The permeability barrier of the skin which prevents transcutaneous water loss and penetration of harmful drugs from the environment is localized in the horny layer of the epidermis. Multiple lipid bilayers obstructing the intercellular space of the stratum corneum fulfill this function. In contrast to cellular membranes consisting predominantly of phospholipids, these lamellae contain mostly ceramides, cholesterol and free fatty acids. The lamellae are derived from the contents of lamellar granules (LGs) which are synthesized in the viable epidermal layers by the keratinocytes. LGs display stacks of small disks each of which represents a flattened vesicle or liposome. Prior to terminal differentiation, the disks are exocytosed into the intercellular space and fused to form uninterrupted sheetlike lamellae. The singular lipid composition of LG-disks and of stratum corneum-lamellae reflects the multistage process of barrier formation. It also renders these structures well suited to provide for a barrier function.

Animals↗
PubMed1988

Head structure of bacteriophages T2 and T4.

The length-to-width ratios of bacteriophage T2 and T4 heads and stereometric angles specifying the prolate icosahedral T2 capsid were evaluated on electron micrographs recorded from samples prepared by a variety of methods. The copy numbers of the major capsid protein, gp23*, of T2 and T4 phages were compared by quantitative gel electrophoresis. Taken together, the resulting values are most compatible with triangulation numbers T = 13 and Q = 21 for both T2 and T4, thus confirming the previously proposed capsid architecture of T4 revealed by indirect measurements and thereby eliminating the repeatedly reported discrepancy between T2 and T4 in favor of a common Q number of 21 corresponding to 960 copies of gp23*.

Capsid↗
PubMed1987

Stratum corneum lipid liposomes: calcium-induced transformation into lamellar sheets.

The epidermal water barrier in mammalian stratum corneum is formed of broad lamellar sheets of lipids consisting principally of ceramides (40%), cholesterol (25%), cholesteryl sulfate (10%), and free fatty acids (25%). Such lipid mixtures have been shown to form lipid bilayers in the form of small, unilamellar liposomes when sonicated at 80 degrees C in water containing Tris buffer and 100 mM NaCl. In the present study it is shown that such liposomes are slowly transformed into large unilamellar liposomes and then into broad lamellar sheets after the addition of stoichiometric amounts of calcium chloride. The presence of free fatty acids was a necessary condition for this calcium-induced fusion. These observations may provide a useful analogy for the transformation of flattened liposomes into broad lamellar sheets that occurs during transition of epidermal granular cells into corneocytes.

Animals↗
PubMed1986

Preparation of liposomes from stratum corneum lipids.

Mammalian stratum corneum contains multiple intercellular lipid bilayers that constitute the epidermal water barrier. Unlike all other biologic membranes, the epidermal lamellae do not contain phospholipids, as a result of which the ability of the stratum corneum lipid mixture to form bilayers has been questioned. In the present study, a lipid mixture containing only epidermal ceramides (40%), cholesterol (25%), palmitic acid (25%), and cholesteryl sulfate (10%), approximating the composition of stratum corneum lipids, formed stable, unilamellar liposomes when sonicated at 80 degrees C in buffer containing 100 mM NaCl, 5 mM Tris, and 1 mM EDTA at pH 7.5. The size and form of the liposomes were studied by both freeze fracture and negative staining electron microscopy. Lipid mixtures from which either the palmitic acid or the cholesteryl sulfate were omitted were still capable of forming similar liposomes, but a mixture of ceramides and cholesterol, or ceramides alone, were incapable of forming liposomes. The results indicate that lipid mixtures similar to those found in stratum corneum are capable of forming bilayers at physiologic pH.

Animals↗