The Ca2+ channel β2 subunit is selectively targeted to the axon terminals of supraoptic neurons

The assembly of high voltage-activated Ca²⁺ channels with different β subunits influences channel properties and possibly subcellular targeting. We studied β subunit expression in the somata and axon terminals of the magnocellular neurosecretory cells, which are located in the supraoptic nucleus (SON) and neurohypophysis, respectively. Antibodies directed against the 4 Ca_Vβ subunits (Ca_Vβ₁-Ca_Vβ₄) were used for immunoblots and for immunostaining of slices of these two tissues. We found that all 4 β subunits are expressed in both locations, but that Ca_Vβ₂ had the highest relative expression in the neurohypophysis. These data suggest that the Ca_Vβ₂ subunit is selectively targeted to axon terminals and may play a role in targeting and/or regulating the properties of Ca²⁺ channels.     

Uric acid modulates vascular endothelial function through the down regulation of nitric oxide production

Abstract

Endothelial dysfunction characterized by decreased nitric oxide (NO) bioavailability is the first stage of coronary artery disease. It is known that one of the factors associated with an increased risk of coronary artery disease is a high plasma level of uric acid. However, causative associations between hyperuricaemia and cardiovascular risk have not been definitely proved. In this work, we tested the effect of uric acid on endothelial NO bioavailability. Electrochemical measurement of NO production in acetylcholine-stimulated human umbilical endothelial cells (HUVECs) revealed that uric acid markedly decreases NO release. This finding was confirmed by organ bath experiments on mouse aortic segments. Uric acid dose-dependently reduced endothelium-dependent vasorelaxation. To reveal the mechanism of decreasing NO bioavailability we tested the effect of uric acid on reactive oxygen species production by HUVECs, on arginase activity, and on acetylcholine-induced endothelial NO synthase phosphorylation. It was found that uric acid increases arginase activity and reduces endothelial NO synthase phosphorylation. Interestingly, uric acid significantly increased intracellular superoxide formation. In conclusion, uric acid decreases NO bioavailability by means of multiple mechanisms. This finding supports the idea of a causal association between hyperuricaemia and cardiovascular risk.     

Adaptation to salinity at the plant cell level

Summary Various mechanisms of adaptation of plant cells to salinity are reviewed: (1) protection of enzymes and maintenance of turgor by organic solutes; (2) prevention of ion toxicity by compartmentation; and (3) energization of solute transport by the proton pump. All these mechanisms seem to play a role in adaptation. The particular advantages of using salt-adapted cells in suspension culture to identify mechanisms of adaptation are pointed out.     

Development of membrane-potential responsiveness by myeloid leukemia cells during neutrophilic differentiation

The ability of a myeloid leukemia cell line (HL-60) to undergo membrane electrical potential changes was followed during neutrophilic differentiation induced by 2 compounds. Membrane-potential changes were induced with 12-O-tetradecanoylphorbol 13-acetate (TPA) or formyl-methionyl-leucyl-phenylalanine (FMLP) and were monitored by flow cytometry. The magnitude of the membrane-potential response to TPA increased in a more uniform manner as the population of cells matured than did acquisition of mature morphology or ability to undergo the respiratory burst in response to TPA. The response to TPA and FMLP of HL-60 cells, maximally induced to differentiate by dimethylsulfoxide, closely resembled that of neutrophils. Thus, HL-60 cells may be a useful tool in the study of the relation between membrane depolarization and subsequent cellular activation.     

The insertion of mesenchyme cells into the ectoderm during differentiation in Sea urchin embryos

Summary During the course of sea urchin development, from early blastula to pluteus larva, there are two major visible processes toward which all activities seem to be focused. They are the differentiation of the larval skeleton by the primary mesenchyme cells and the differentiation of the primitive gut by the secondary mesenchyme cells. These activities take place within the shell-like layer of epithelial cells, or ectodermal wall. The interactive role of the ectodermal wall with the mesenchyme cells is not yet clearly understood. A number of earlier studies have proposed that the ectoderm may have an inductive influence on the mesenchyme cells and that its inner surface forms a molecular template for guiding the mesenchyme cells. In this report, we suggest an additional role for the ectodermal wall. We show that some primary mesenchyme cells and secondary mesenchyme cells insert between the cells of the ectodermal wall in order to firmly anchor the anlage of the larval skeleton and primitive gut during differentiation. This mechanism may provide a physical basis for maintaining the stable positional relationship of the anlage during development.     

Chloride cell responses to long-term exposure to distilled and hard water in the gill of the armored catfish, Hypostomus tietensis (Loricariidae)

The morphological changes in the gill chloride cells of the armored catfish, Hypostomus tietensis , were investigated after 15 days' exposure to either distilled or hard water. The thickness of the water–blood barrier in the lamellae increased significantly in fish kept in distilled water due to the high proliferation of chloride cells. The apical surface of about 68% of chloride cells was sharply reduced by the development of an apical crypt with a sponge-like surface, although no change in the chloride cell fractional area was found. In contrast, H. tietensis kept in Na⁺, Cl⁻ and Ca²⁺ rich water displayed no significant changes in the number of chloride cells or in their apical surface morphology compared with the control fish. Chloride cell response to ion challenge in H. tietensis suggested the involvement of different strategies to maintain homeostasis in ion-poor water, which may be related to the life history of species.     

Mercury in blood cells—Altered elemental profiles

The study was composed of 27 persons that displayed vague symptoms similar to those of the victims of Minamata and Iraq. Skew distributions of mercury were observed in individual erythrocytes and neutrophil granulocytes when measured by PIXE. Mercury could not be detected in the platelets. The erythrocytes also displayed lowered concentrations of magnesium and zinc, together with increased concentrations of calcium and strontium. The neutrophils displayed decreased concentrations of magnesium and zinc and increased concentrations of calcium, strontium, and iron. The presence of mercury and the altered elemental profiles in the erythrocytes and the neutrophil granulocytes are suggested as early signs of exposure.     

A simple system for plant cell microinjection and culture

Microinjection of plant protoplasts and cells has been recently reported, however a system that combines simplicity of design, harmless immobilization, high resolution visibility and ability to monitor individual target cells is lacking. This report describes a system which combines these features. It consists of a microinjection-microculture dish containing immobilized protoplasts and a simple chamber that maintains sterility and humidity during injection. Highly purified protoplast preparations are plated at low population density as a thin monolayer of widely separated cells embedded in agarose layered over a thicker (0.2 mm at center to 1 mm at edge) layer of agarose-solidified medium. This physical arrangement allows for rapid location, mapping and injection of the immobilized protoplasts and also their subsequent location for growth monitoring. The double layers of agarose provide adequate nutrition for culturing injected cells to the microcalli stage. In addition to protoplast injection, this system was also used to inject 3- to 4-day old nonspherical cells derived from protoplasts. Colony formation rates from injected protoplasts and cells with regenerated walls were equivalent to those of uninjected controls. Furthermore, tobacco protoplasts stored at 4°C in liquid medium for up to two weeks remained fully competent for plating and injection. These cold-stored protoplasts, when injected, formed colonies at rates similar to those from fresh preparations. The ability to store protoplasts without loss of viability considerably increases the ease and convenience of cell injection experiments.Mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by the USDA, and does not imply its approval to the exclusion of the other products that may also be suitable.