Role of apoplast acidification by the H+ pump

We have studied the mechanism of the response to iron deficiency in rape (Brassica napus L.), taking into account our previous results: net H⁺ extrusion maintains a pH shift between the root apoplast and the solution, and the magnitude of the pH shift decreases as the buffering power in the solution increases. The ferric stress increased the ability of roots to reduce Fe[III]EDTA. Buffering the bulk solution (without change in pH) inhibited Fe[III]EDTA reduction. At constant bulk pH, the inhibition (ratio of the Fe[III]EDTA-reduction rates measured in the presence and in the absence of buffer) increased with the rate of H⁺ extrusion (modulated by the length of a pretreatment in 0.2 mM CaSO₄). These results support the hypothesis that the apoplastic pH shift caused by H⁺ excretion stimulated Fe[III] reduction. The shape of the curves describing the pH-dependency of Fe[III]EDTA reduction in the presence and in the absence of a buffer fitted this hypothesis. When compared to the titration curves of Fe[III]citrate and of Fe[III]EDTA, the curves describing the dependency of the reduction rate of these chelates on pH indicated that the stimulation of Fe[III] reduction by the apoplastic pH shift due to H⁺ excretion could result from changes in electrostatic interactions between the chelates and the fixed chargers of the cell wall and-or plasmalemma. Blocking H⁺ excretion by vanadate resulted in complete inhibiton of Fe[III] reduction, even in an acidic medium in which there was neither a pH shift nor an inhibitory effect of a buffer. This indicates that the apoplastic pH shift resulting from H⁺ pumping is not the only mechanism which is involved in the coupling of Fe[III] reduction to H⁺ transport. Our results shed light on the way by which the strong buffering effect of HCO ₃ ^- in some soils may be involved in iron deficiency encountered by some of the plants which grow in them.     

A model for predicting ionic equilibrium concentrations in cell walls

Purified cell walls were prepared from roots of Horse bean (Vicia faba L., var. minor) and Yellow Lupine (Lupinus luteus L.). Two methods were used: (a) grinding and (b) lysis of the endocellular contents by Triton X-100. The Ca²⁺, Mg²⁺, and K⁺ contents were determined after incubation in various solutions in such a manner that the measurements gave access to the undisturbed equilibrium contents. The results were used to test a model which describes the ionic atmosphere in the cell walls as a result of simultaneous electrostatic interactions between free ions (Donnan effect) and specific association equilibria, including acido-basic ones. This model correctly accounted for the whole set of experimental results and predicted the values of the unmeasurable local concentrations and pH.     

Purification of androgen-binding protein from rat testis using high-performance liquid chromatography and physicochemical properties of the iodinated molecule

The androgen-binding protein (ABP) has been purified 87,500-fold from rat testis using 4 steps of HPLC, with a yield of 14%. The molecule was 99% pure with a specific activity estimated to 16,600 pmol/mg protein. The iodinated molecule was eluted in 2 peaks in Sephacryl S300 gel filtration with a molecular mass estimated to be 92,600 +/- 3300 and 50,300 +/- 4000 Da. The column isoelectrofocusing of 125I-ABP demonstrated 3 isoproteins isoelectric at pH 4.7, 4.9 and 5.3 and the sedimentation coefficient was estimated to be 4.7 S in sucrose gradient ultracentrifugation. The 125I-ABP had similar physiochemical properties to the non-labelled ABP of epididymis.     

Evidence of density-independent mortality in a settling coral reef damselfish, Chromis viridis

To know if the variation in the number of settling fish larvae can be dampened by density-dependent postsettlement mortality, we investigated the relationship between settler density and predator-induced mortality of a coral reef damselfish, Chromis viridis. Totals of 2, 3, 5, 8, 10, 12, 14, 16, 18, and 20 fish of 10 or 20 mm total length were released in experimental cages enclosing a coral head of Porites rus (to provide settlement habitat) and five predators. The results showed that the mortality rate of both 10- and 20-mm fish was density independent.     

Recent advances on proteins of plant terminal membranes.

Since the beginning of the 1990s, our knowledge of the protein equipment of plant membranes progresses at an accelerating pace, owing to the irruption of molecular biology tools and genetics strategies in plant biology. Map-based cloning strategies and exploration of EST databases rapidly enrich the catalog of cDNA or gene sequences expected to code for membrane proteins. The accumulation of 'putative' membrane proteins reinforces the need for structural, functional and physiological information. Indeed, ambiguities often exist concerning the association to a membrane, the membrane identity and the topology of the protein inserted in the membrane. The combination of directed mutagenesis and heterologous expression of plant genes in various systems and plant reverse genetics has opened the possibility to study molecular and physiological functions. This review will emphasize how these tools have been essential for the exciting recent discoveries on plant terminal membrane proteins. These discoveries concern a variety of transport systems for ions, organic solutes including auxin, water channels, a large collection of systems suspected to act as receptors of chemical signals, proteins thought to control vesicle trafficking and enzymatic systems.     

Ultrastructure des capillaires dans le télencéphale du poulet au cours de l'embryogenèse et de la croissance postnatale

Resumé Une étude en microscopie électronique de la structure des capillaires et des formations cellulaires qui les entourent a été faite au niveau du télencéphale, chez le Poulet, au cours du développement embryonnaire et de la croissance postnatale.L'endothélium capillaire est présent au premier stade embryonnaire étudié (embryon agé de 8 jours). La membrane basale se différencie vers le dix-neuvième jour du développement embryonnaire, et elle est précédée de l'apparition des péricytes qui restent inclus définitivement dans sa structure.L'espace extracellulaire dans l'environnement des capillaires, constitué par de larges lacunes au cours du développement embryonnaire, se réduit dès l'éclosion à des intervalles de 150–200 Å de diamètre. Les astrocytes et leurs prolongements (pieds vasculaires) atteignent leur complet développement, et entourent totalement le capillaire, vers le vingtième jour de la croissance postnatale.Nos observations mettent en évidence la relation étroite existant entre le développement des structures gliales péricapillaires et l'apparition de certains mécanismes du système de la barrière hémo-encéphalique chez le Poulet.

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Ultrastructure of the capillaries in the developing telencephalon of the chicken

Summary An electron microscopical study was made of the structure of the capillaries and of the surrounding cell formations in the chick telencephalon, during embryonic development and postnatal growth.The capillary endothelium is present in the embryonic stage first studied (i. e. the 8-day old embryo). The basement membrane differentiates about the nineteenth day of embryonic development, and it is preceded by the appearance of the pericytes which are finally included in its structure.The extracellular space in the vicinity of the capillaries, made of large gaps during the embryonic development, is confined from hatching time to intervals 150–200 Å diameter. The astrocytes and their processes (vascular feet) reach their complete development, and entirely surround the capillaries, on about the twentieth day of the postnatal growth.Our observations reveal the close connection between the development of the pericapillary glial structures and the appearance of some mechanisms of the blood-brain barrier system in the chick.

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Les auteurs remercient Madame R. Hatier de son excellente collaboration technique.     

Histoimmunological identification of a prolactin-like substance in rodent testis

Summary Anti-rat prolactin (PRL) antibodies were localized by histoimmunological methods in the cytoplasm of testicular interstitial cells, Sertoli cells, spermatogonia and primary spermatocytes of the rat and mouse. Control of specificity by affinity chromatography methods showed this PRL-like material to be non-specific in these testicular tissues, but specific in adenohypophyseal cells. These results are discussed.     

Immunolocalization of some plasmatic proteins in basement membranes during earliest rat morphogenesis with special reference to the gonadal differentiation.

Rat albumin, transferrin, angiotensinogen and T kininogen were examined immunohistochemically in the epithelial basement membranes (BMs) during the earliest rat morphogenesis. As a specific marker for BMs, laminin was used. Albumin and transferrin immunostaining appeared as early as the 11th day of gestation in all epithelial BMs. In 13-day-old mesonephric-gonadal complex, just after the onset of the sexual cord differentiation, all BMs were weakly stained. One day later, a stronger immunoreactivity was distributed along the coelomic epithelium, the Wolffian duct, the mesonephric tubules, the differentiating sexual cords and the blood vessels. The epidermal BM and all epithelial BMs of differentiating organs are also immunoreactive. The accumulation of albumin and transferrin in the BMs is probably the result of a strong release of these two major liver proteins in the embryonic blood and their diffusion in extracellular spaces. At these stages, the lack of angiotensinogen and T kininogen BM labeling is consistent with their low hepatic and plasmatic concentrations. During embryogenesis, some plasma proteins are probably trapped in the epithelial BMs and not produced by local cells.     

Cd-induced growth reduction in the halophyte <Emphasis Type="Italic">Sesuvium portulacastrum</Emphasis> is significantly improved by NaCl

The effects of Cd²⁺ and NaCl, applied together or separately, on growth and uptake of Cd²⁺ were determined for the halophyte Sesuvium portulacastrum L. Seedlings were cultivated in the presence of 50 or 100 μmol L⁻¹ Cd²⁺ alone or combined with 100 or 400 mmol L⁻¹ NaCl. Data showed that alone, Cd²⁺ induced chlorosis, necrosis, and inhibited growth. Addition of NaCl to Cd²⁺-containing medium restored growth and alleviated the toxicity, however. NaCl also enhanced the amounts of Cd²⁺ accumulated in the shoots. All Cd²⁺ treatment reduced K⁺ and Ca²⁺ uptake and transport to the shoots. Accumulation of Na⁺ in the shoots was not affected by Cd²⁺, however. Thus S. portulacastrum maintained its halophytic characteristics in the presence of Cd²⁺. We suggest this halophyte could be used for phytoextraction of Cd²⁺ from salt-contaminated sites.     

Effect of pH on Orthophosphate Uptake by Corn Roots

Orthophosphate (Pi) influx in washed corn roots was studied with experimental conditions allowing a distinction of pH effects on Pi ionization in the medium and on the transport system itself. There appeared to be no relationship between the pH dependencies of membrane potential, H⁺ secretion, and ³²Pi influx. The Pi uptake versus pH curves were compared to the calculated ones describing the concentrations of the different ionized Pi forms in the medium and in the cell walls; the latter were obtained using the theoretical model described by Sentenac and Grignon (1981) Plant Physiol 68: 415-419). The conclusion was that the transported form is H₂PO₄⁻ and the concentration sensed by the transport system is the local one. The ionic compositions of experimental media were manipulated to ensure constant pH and various H₂PO₄⁻ concentrations, or constant H₂PO₄⁻ concentration and various pH values in the walls. The kinetic analysis of the results in the micromolar range showed that the transport system has an intrinsic sensitivity to pH, and is switched from a low activity state at pH > 6 to a high activity one at pH < 4 (pH in the walls). This change could be triggered by the protonation of a group with pK 5.5.