Specialized formations of endoplasmic reticulum in parenchyma cells ofMimosa pudica L.

Summary Parenchyma cells ofMimosa pudica display close associations between two or more cisternae of the endoplasmic reticulum. These associations form simplified types of lamellar bodies in which inner paired lamellae have lost their ribonucleoprotein granules and are separated by a dense layer.     

The dopaminergic innervation of the goldfish pituitary

Summary The dopaminergic innervation of the goldfish pituitary gland was studied by immunocytochemistry at the electron-microscope level using highly specific antibodies against dopamine coupled to bovine serum albumin with glutaraldehyde. A satisfactory preservation of the tissue was achieved after immersion in 5% glutaraldehyde in phosphate buffer containing sodium metabisulfite to prevent oxidation of the endogenous dopamine. The immunocyto-chemical procedure was performed on Vibratome sections using the preembedding method. Immunoreactivity was restricted to part of the neurosecretory type-B fibers (diameter of the secretory vesicles lower than 100 nm) in which it was found to occupy the whole cytoplasm. Labeled fibers were observed within the neurohypophysis in the different parts of the gland and in the adenohypophyseal tissue where immunoreactive profiles were detected in close apposition to the different cell types. These data are in agreement with previous results obtained by means of radioautography and further support a role for dopamine in the neuroendocrine regulation of pituitary functions in teleosts.     

Dimethylsulfoxide action on dark- and light-induced leaflet movements and its necrotic effects on excised leaves of Cassia fasciculata

Dimethylsulfoxide (DMSO) acts on dark- and light-induced movements exhibited by leaflets of isolated leaves of Cassia fasciculate Michx. The closing movement (scotonasty), induced when the leaves are placed in darkness during the normal period of daylight, was inhibited, whereas the opening movement (photonasty), when the leaves arc transferred to light during the normal period of darkness, was promoted. The concentration for significant effects of DMSO was 1% (v/v) when applied over a 3-h period. After five days, a necrosis of the leaflets was observed for DMSO concentrations as small as 0.1%, applied over a 6-h period. Complete abscission took place if 3% DMSO was applied for more than 30 min.     

Immunolocalization of the Plasma Membrane H+ -ATPase in Minor Veins of Vicia faba in Relation to Phloem Loading

The phytotoxin fusicoccin (FC), after binding to a plasma membrane-localized receptor, causes higher plant cells to excrete protons. Ligand-binding analysis has been used to show that the plasma membrane of mung bean (Vigna radiata L.) hypocotyls contains both high-affinity (HA) and low-affinity (LA) binding sites for FC. The effect of tissue maturation on these sites was determined on isolated membrane vesicles from the meristematic region (hook) and the elongation zone and from mature hypocotyl tissues. In the meristematic region the HA:LA ratio was 1:20. As hypocotyl tissues matured, the site density of HA increased and there was no change in LA density, so that the HA:LA ratio increased to 1:2 in maturet issues. FC-induced proton excretion correlates with the HA density, not the LA density. When sections isolated from each region were incubated with FC prior to isolation of membranes, there was an apparent conversion of LA to HA sites during the first 90 min in all regions. During the next 1 to 3 h there was a further 2.5- to 3- fold increase in binding sites in all regions, accompanied by a slight decline in dissociation constant. The increase in binding sites, but not the apparent conversion of LA to HA, was partly blocked by cycloheximide. These data suggest that FC alters FC-binding protein activity in two ways: first, by causing an increase in affinity for FC of preexisting LA receptors, and second by inducing the synthesis of additional FC receptors. This apparent up-regulation of a phytotoxin receptor by its ligand in plants has not previously been reported.     

Absence of plasma membrane H+-ATPase in plasmodesmata located in pit-fields of the young reactive pulvinus ofMimosa pudica L.

Summary Immunocytochemical techniques were employed to study the spatial distribution of the plasma membrane H⁺-ATPase within various cell types of the young reactive primary pulvinus ofMimosa pudica L. These cells were interconnected by large numbers of plasmodesmata, being concentrated within pit-fields. Although we could routinely detect evidence of the H⁺-ATPase along the plasma membrane, immunolabelling was rarely, if ever, observed along the plasma membranes of the plasmodesmata. This finding is discussed with respect to the likely specialized supramolecular structure of the plasmodesma.Abbreviations SEL size exclusion limit of plasmodesmata     

Control of Vascular Sap pH by the Vessel-Associated Cells in Woody Species (Physiological and Immunological Studies)

In Robinia wood, the vessel-associated cells form a continuous sleeve around the vessels. Variations in pH of the solution perfused through the vessels during the annual cycle and the opposing effects of carbonyl cyanide-m-chlorophenylhydrazone and fusicoccin on this pH value indicate that some living cells of the wood are involved in the control of vascular sap pH and that this control fluctuates with the seasons. The immunolocalization of the plasma membrane HT+-ATPase in Robinia wood was studied by the immunogold-silver-staining technique using an antibody raised against a conserved stretch of the cytoplasmic domain of the H+-ATPase. The immunostaining is much stronger in vessel-associated cells than in other living cell types (ray and axial parenchyma elements) of the secondary xylem. Our data show an efficient involvement of this cell type in the control of vascular sap pH.     

Studies on Possible Mechanisms of Early Functional Compensatory Adaptation in the Remaining Kidney

Two to 4 hours after unilateral renal exclusion in rats, urine flow rate from the remaining kidney had increased to twice the control level, whereas the filtration rate remained unchanged. After contralateral nephrectomy, NGFR was similar to that of controls, but fractional water reabsorption along proximal tubules decreased. Protein concentration in efferent arteriolar plasma, and hydrostatic pressure gradient between proximal tubules and peritubular capillaries were similar in experimental and control kidneys. Unilateral renal exclusion was followed by a rapid increase of blood pressure. Prevention of this rise depressed but did not abolish functional compensatory adaptation. The occurrence of compensatory adaptation was not affected by decreased renal perfusion pressure.     

Influence de la teneur en oxygène sur la sensibilité de certains stades juvéniles deSitophilus oryzae etS. granarius au dioxyde de carbone

Résumé Certains stades juvéniles de charançons des céréales du genreSitophilus ont été soumis à un séjour prolongé dans des compositions gazeuses modifiées par le dioxyde de carbone (CO₂).Les auteurs se sont placés dans des conditions de teneur en CO₂ constante (50%), associée à des teneurs en oxygène comprises entre 4 et 20%, pour étudier les conditions d'une synergie entre l'effet spécifique du CO₂ et la présence d'une forte teneur résiduelle en oxygène. Les effets ont été observés sur les stades juvéniles les plus évolués (larves et nymphes) des espècesS. oryzae (L.) etS. granarius (L.), (Coleoptera: Curculionidae).L'accroissement de la vitesse de mortalité chezS. oryzae est significative avec l'augmentation de la pression partielle d'oxygène. Cet effet de synergie provoqué par l'oxygène n'est pas significatif avecS. granarius, bien qu'il existe aussi avec cette espèce une tendance à l'amélioration de l'efficacité insecticide à court terme avec les mélanges à forte teneur résiduelle en oxygène.Il en est déduit des hypothèses sur les effets des mélanges gazeux à composition modifiée par le CO₂ au niveau de la physiologie générale de ces insectes, qui est très difficile à appréhender directement à cause du mode de développement des stades juvéniles des charançons (formes cachées dans le grain).     

Revisiting the Central Metabolism of the Bloodstream Forms of Trypanosoma brucei: Production of Acetate in the Mitochondrion Is Essential for Parasite Viability

Background

The bloodstream forms of Trypanosoma brucei, the causative agent of sleeping sickness, rely solely on glycolysis for ATP production. It is generally accepted that pyruvate is the major end-product excreted from glucose metabolism by the proliferative long-slender bloodstream forms of the parasite, with virtually no production of succinate and acetate, the main end-products excreted from glycolysis by all the other trypanosomatid adaptative forms, including the procyclic insect form of T. brucei.

Methodology/Principal Findings

A comparative NMR analysis showed that the bloodstream long-slender and procyclic trypanosomes excreted equivalent amounts of acetate and succinate from glucose metabolism. Key enzymes of acetate production from glucose-derived pyruvate and threonine are expressed in the mitochondrion of the long-slender forms, which produces 1.4-times more acetate from glucose than from threonine in the presence of an equal amount of both carbon sources. By using a combination of reverse genetics and NMR analyses, we showed that mitochondrial production of acetate is essential for the long-slender forms, since blocking of acetate biosynthesis from both carbon sources induces cell death. This was confirmed in the absence of threonine by the lethal phenotype of RNAi-mediated depletion of the pyruvate dehydrogenase, which is involved in glucose-derived acetate production. In addition, we showed that de novo fatty acid biosynthesis from acetate is essential for this parasite, as demonstrated by a lethal phenotype and metabolic analyses of RNAi-mediated depletion of acetyl-CoA synthetase, catalyzing the first cytosolic step of this pathway.

Conclusions/Significance

Acetate produced in the mitochondrion from glucose and threonine is synthetically essential for the long-slender mammalian forms of T. brucei to feed the essential fatty acid biosynthesis through the “acetate shuttle” that was recently described in the procyclic insect form of the parasite. Consequently, key enzymatic steps of this pathway, particularly acetyl-CoA synthetase, constitute new attractive drug targets against trypanosomiasis.     

A Trypanosoma brucei Kinesin Heavy Chain Promotes Parasite Growth by Triggering Host Arginase Activity

Background

In order to promote infection, the blood-borne parasite Trypanosoma brucei releases factors that upregulate arginase expression and activity in myeloid cells.

Methodology/Principal findings

By screening a cDNA library of T. brucei with an antibody neutralizing the arginase-inducing activity of parasite released factors, we identified a Kinesin Heavy Chain isoform, termed TbKHC1, as responsible for this effect. Following interaction with mouse myeloid cells, natural or recombinant TbKHC1 triggered SIGN-R1 receptor-dependent induction of IL-10 production, resulting in arginase-1 activation concomitant with reduction of nitric oxide (NO) synthase activity. This TbKHC1 activity was IL-4Rα-independent and did not mirror M2 activation of myeloid cells. As compared to wild-type T. brucei, infection by TbKHC1 KO parasites was characterized by strongly reduced parasitaemia and prolonged host survival time. By treating infected mice with ornithine or with NO synthase inhibitor, we observed that during the first wave of parasitaemia the parasite growth-promoting effect of TbKHC1-mediated arginase activation resulted more from increased polyamine production than from reduction of NO synthesis. In late stage infection, TbKHC1-mediated reduction of NO synthesis appeared to contribute to liver damage linked to shortening of host survival time.

Conclusion

A kinesin heavy chain released by T. brucei induces IL-10 and arginase-1 through SIGN-R1 signaling in myeloid cells, which promotes early trypanosome growth and favors parasite settlement in the host. Moreover, in the late stage of infection, the inhibition of NO synthesis by TbKHC1 contributes to liver pathogenicity.