Modulation of the membrane potential of the Schwann cell of the squid giant nerve fiber

1. The involvement of second messengers and of other chemical mediators, in the modulation of the membrane potential of the Schwann cell of the giant nerve fiber of the Tropical squid Sepioteuthis sepioidea is described. 2. The involvement of the cyclic nucleotide adenosine 3', 5' monophosphate (cAMP) in mediating the actions of the nicotinic Ach receptors of the Schwann cells is suggested. 3. The presence of octopaminergic receptors in the Schwann cells, mediating their actions through the activation of adenylate cyclase, is also described. 3. Receptors for vasoactive intestinal peptide (VIP) are also present on the Schwann cells, and their actions are mediated via a second messenger system that does not involve the activation of adenylate cyclase. 5. The three independent receptor systems referred above are able to interact in a complex way, which involves both their direct actions on the Schwann cell membrane potential and modulatory effects between the systems.     

Protein composition of the chromosomal scaffold and interphase nuclear matrix

Residual protein structures were prepared from isolated chromosomes and interphase nuclei of in vitro cultured bovine liver cells and the protein compositions were analysed. Chromosomes with minimal cytoplasmic contamination were obtained by a simple procedure using a pH 8 isolation medium containing Triton X-100 and polyamines, and residual protein-DNA complexes were prepared by extraction with 2 M NaCl. Residual protein structures were also obtained by digesting isolated chromosomes with staphylococcal nuclease. Protein compositions of both structures as obtained by SDS-polyacrylamide gel electrophoresis were essentially the same. Residual protein structures were prepared from isolated nuclei by the same procedures. The major nuclear matrix proteins, i.e., the lamins A, B, and C, were not found in the chromosomes and chromosome scaffolds. On the other hand, the residual chromosome structures contained two major polypeptides of 37 and 83 kilodalton relative molecular weights that were absent from the nuclear matrix preparations. A few polypeptides with the same or very similar electrophoretic mobilities were found in the residual structures of both the nuclei and the chromosomes.     

Die Diffusion des 59Fe-markierten Hämoglobins,ein Artefakt der Glutaraldehyd-Fixierung

Zusammenfassung Ratten wurden 50–80 c ⁵⁹Fe-Citrat in die caudale Vene injiziert. Nach 3–5tägigem Einbau des markierten Eisens in das Hämoglobin wurden Milz-Blöckchen (2 × 2 × 5 mm) 2 Std in GA vorfixiert oder in Hanks-Lösung (= Kontrolle) überführt. Ein Teil der Blöckchen wurde anschließend in OsO₄-Lösung nachfixiert.Die autoradiographischen Ergebnisse zeigen eine Diffusion des Hämoglobins vom unfixierten Zentrum zur Peripherie des Blöckchens.Aktivitätsbestimmungen, die am ganzen Blöckchen, dessen abgetrennten zentralen und peripheren Anteilen, sowie im Überstand vorgenommen wurden, bestätigen diese Diffusion. Während der OsO₄-Nachfixierung erfolgte ein weiterer Verlust des markierten Hämoglobins aus dem zentralen Teil des Blöckchens, nicht aber aus der vorfixierten Peripherie.

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The diffusion of ⁵⁹Fe-labelled hemoglobin, an artefact of the fixation with glutaraldehyde

Summary Injections of 50–80 c ⁵⁹Fe-citrate into the caudale vein of rats were performed. After 3–5 days of ⁵⁹Fe incorporation into the hemoglobin the spleen was taken off, cut in small blocks (2 × 2 × 5 mm) and prefixed for 2 hours in GA or transfered into Hankssolution (= control). Later on some blocks of the spleen were postfixed in OsO₄ solution.A diffusion of the hemoglobin from the unfixed center to the peripheral tissue of the spleen-block is demonstrated by autoradiographic results.After measuring the radioactivity of the total spleen-block of the separated central and peripherical parts as well as their supernatants a diffusion was confirmed. A further loss of the labelled hemoglobin has been observed during the OsO₄ postfixation from the central part of the spleen-block, but not from the prefixed periphery.

     

Leukocyte inhibitory factor activates human neutrophils and macrophages to release leukotriene B4 and thromboxanes

Recent evidence has proved that cytokines can stimulate the production of 5-lipoxygenase products. Leukotriene B4 (LTB4) is a major mediator of leukocyte activation in acute inflammatory reactions, which produce chemotaxis, lysosomal enzyme release, and cell aggregation. Leukocyte inhibitory factor (LIF) also causes biological responses related to inflammation, i.e., LIF directly induces specific granule secretion by polymorphonuclears (PMNs) and potentiates many formyl-methionyl-leucyl-phenylalanine (FMLPs) mediated responses. Since arachidonic acid products are important mediators of inflammation, we have studied the effects of LIF on the arachidonic acid cascade products LTB4 and thromboxane A2 (TxA2). Resuspended at a final concentration of greater than 95% polymorphonuclear PMNs were isolated and tested with some cytokines on the release of LTB4 and TxA2. Peripheral blood mononuclear cells were isolated and seeded in Petri dishes and incubated for 60 min. Adherent macrophages were used for the cytokine stimulation study. Both types of leukocytes were treated with LIF, interleukin 6 (IL 6), and granulocyte-monocyte colony stimulating factor (GM-CSF) at different concentrations, and test agents A23187 and FMLP. Radioimmunoassay for LTB4 and TxB2 was determined by the resulting supernatants. Treatment of PMNs and macrophages with LIF at different concentrations proved to generate significant increases in LTB4 and TxA2 production. This was compared with IL 6 and GM-CSF, which had no effects. In these experiments, TxA2 generations could not be attributed to platelet contamination of PMN suspensions. The quantity of platelet contamination was not sufficient to influence how much TxB2 was produced. The similarities of LIF to other arachidonate stimulating cytokines suggest a similar mode of action in producing hematologic changes typical of tissue injury.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of oxygen availability and salinity on EARLY LIFE HISTORY STAGES OF SALT MARSH PLANTS. I. Different germination strategies of Spartina alterniflora and Phragmites australis (Poaceae)

Gradients in oxygen availability and salinity are among the most important environmental parameters influencing zonation in salt marsh communities. The combined effects of oxygen and salinity on the germination of two salt marsh grasses, Spartina alterniflora and Phragmites australis, were studied in growth chamber experiments. Germination of both species was initiated by emergence of the shoot and completed by root emergence. Percentage S. alterniflora germination was reduced at high salinity (40 g NaCl/L) and in decreased oxygen (5 and 2.5%). In 0% oxygen shoots emerged, but roots did not. P. australis germination was reduced at a lower salinity (25 g NaCl/L) than S. alterniflora, and inhibited at 40 g NaCl/L and in anoxia. However, a combination of hypoxia (10 and 5% O2) and moderate salinity (5 and 10 g NaCl/L) increased P. australis germination. When bare areas in the salt marsh are colonized, the different germination responses of these two species to combinations of oxygen and salt concentrations are important in establishing their initial zonation. In high salinity wetlands S. alterniflora populates the lower marsh and P. australis occupies the high marsh at the upland boundary.     

The Subunit Composition of a GABAA/Benzodiazepine Receptor from Rat Cerebellum

Abstract: The pentameric subunit composition of a large population (36%) of the cerebellar granule cell GABA_A receptors that show diazepam (or clonazepam)-insensitive [³H]Ro 15-4513 binding has been determined by immunoprecipitation with subunit-specific antibodies. These receptors have α₆, α₁, γ_2S, γ_2L, and β₂ or β₃ subunits colocalizing in the same receptor complex.     

Freeze-fracture study of the junctional complexes of human and rabbit thyroid follicles

Summary Zonulae occludentes, gap junctions and desmosomes have been demonstrated in replicas of freeze-fractured follicular cells of normal human and rabbit thyroid glands. The zonulae occludentes between the human follicular cells are composed of two to eight strands, which completely separate the intercellular space from the follicular lumen. Four to twelve or more strands are visible between the follicular cells of the rabbit thyroid gland.In the meshes of the zonulae occludentes as well as below them, gap junctions are present. They are numerous on the fracture faces of the human follicular cell membranes, but infrequent in those of the rabbit.Aggregates of particles related to desmosomes are found in the deeper meshes of the zonulae occludentes or close to them.     

The glomeruli of the human and the rat kidney studied by freeze-fracturing

Glomeruli isolated from rat and human kidneys were studied using the freeze-fracture technique. Discontinuous zonulae occludentes and gap junctions were found in the replicas of the split plasma membrane of the endothelial cells. A diaphragm across the endothelial pores was not demonstrated. The central layer of the basement membrane, corresponding to the lamina densa described in thin sections, revealed a coarse substructure. A slit membrane between the pedicles of the podocytes was not detectable; however, its position was indicated by the different texture of the replica, which abruptly changed at the transition of the basement membrane to the primary urinary space. Furthermore, at the level of the slit membrane arrays of particles were present within the cleaved membrane of the pedicles, probably representing the attachment points of the slit membrane. Isolated strands of a zonula occludens as well as gap junctions were seen on the split plasma membrane of the podocytes. The mesangial cells could be identified by their contiguity to the endothelial cells and by their numerous gap junctions.     

The NHERF1 PDZ2 domain regulates PKA-RhoA-p38-mediated NHE1 activation and invasion in breast tumor cells

Understanding the signal transduction systems governing invasion is fundamental for the design of therapeutic strategies against metastasis. Na(+)/H(+) exchanger regulatory factor (NHERF1) is a postsynaptic density 95/disc-large/zona occludens (PDZ) domain-containing protein that recruits membrane receptors/transporters and cytoplasmic signaling proteins into functional complexes. NHERF1 expression is altered in breast cancer, but its effective role in mammary carcinogenesis remains undefined. We report here that NHERF1 overexpression in human breast tumor biopsies is associated with metastatic progression, poor prognosis, and hypoxia-inducible factor-1alpha expression. In cultured tumor cells, hypoxia and serum deprivation increase NHERF1 expression, promote the formation of leading-edge pseudopodia, and redistribute NHERF1 to these pseudopodia. This pseudopodial localization of NHERF1 was verified in breast biopsies and in three-dimensional Matrigel culture. Furthermore, serum deprivation and hypoxia stimulate the Na(+)/H(+) exchanger, invasion, and activate a protein kinase A (PKA)-gated RhoA/p38 invasion signal module. Significantly, NHERF1 overexpression was sufficient to induce these morphological and functional changes, and it potentiated their induction by serum deprivation. Functional experiments with truncated and binding groove-mutated PDZ domain constructs demonstrated that NHERF1 regulates these processes through its PDZ2 domain. We conclude that NHERF1 overexpression enhances the invasive phenotype in breast cancer cells, both alone and in synergy with exposure to the tumor microenvironment, via the coordination of PKA-gated RhoA/p38 signaling.     

Specificity of a DNA-based (DNAzyme) peroxidative biocatalyst

A complex formation between hemin and a congruous oligonucleotide not only greatly enhances the former’s peroxidative activity but also results in a biocatalyst (DNAzyme) with a novel specificity. Herein substrate, regio-, enantiomeric, and diastereomeric selectivities of heme, the DNAzyme, and the enzyme horseradish peroxidase are comparatively examined.