Metal chelatorNNN’ N’-tetrakis-(2-pyridylmethyl)ethylene diamine inhibits the induction of heat shock protein 70 synthesis by heat in cultured keratinocytes

Heat shock protein (HSP) synthesis results from various types of injury, including heat shock (HS) and some oxidants. The intracellular signals leading to HSP synthesis are not yet fully elucidated. We have studied the influence ofNNN’N’-tetrakis(2-pyridylmethyl)ethylene diamine (TPEN), a metal chelator known to induce cellular zinc and copper deprivation, on resistance to heat and on hsp70 synthesis in HaCaT keratinocytes. TPEN was shown to sensitize HaCaT cells to heat shock. The effect of TPEN was neutralized by equimolar Zn²⁺. By the use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and Western blotting characterization of hsp70, it was shown that cultured HaCaT cells constitutively express the inducible form of hsp70. The application of TPEN alone slightly increases the level of hsp70 but inhibits its induction by HS. This inhibitory effect is related to metal deprivation, because it is eliminated when Cu²⁺ or Zn²⁺ ions are supplied together with TPEN. These results suggest that these metals are involved in the expression by keratinocytes of a stress protein which has a protective action against environmental stress.     

Quantification of the effects of Septoria tritici blotch on wheat leaf gas exchange with respect to lesion age, leaf number, and leaf nitrogen status

Quantification of the damaging effects of pathogens on diseased plants and inclusion of these damaging functions in crop simulation models is of great importance for a more complete understanding of yield response to diseases. In this study, the effect of Septoria tritici blotch (STB) on net photosynthetic and dark respiration rates of wheat flag leaves was quantified. Bastiaans' model: Y=(1-x)beta was used to characterize the relationship between relative leaf photosynthesis (Y, considering Ynet and Ygross) and STB severity (with x the proportion of the diseased area). The value of beta indicates whether the effect of disease on photosynthesis is larger (beta >1), lower (beta <1), or equal (beta =1) to the proportion of visible diseased area. In the experimental conditions used here, leaf nitrogen content (in a range from 0.18 to 0.24 mg cm(-2)), and leaf number (flag and second leaves) did not significantly influence the effect of STB on leaf gas exchange. By contrast, damage depended strongly on the developmental stages of the STB lesions. STB lesions had no effect on inoculated leaves before visible symptoms appeared. Chlorotic symptoms had less effect on leaf net photosynthetic rate than could be accounted for by the visible diseased area (betanet=0.81). The effect of necrotic lesions on the leaf net photosynthetic capacity was slightly greater than that accounted for by visible symptoms (betanet=1.35). Our results suggest that the effect of the necrotic symptoms on the net photosynthesis expressed by betanet >1 is due to a combination of a decrease in the gross photosynthesis (betagross still >1) and to an increase in the dark respiration rate (betagross相似文献   

The Lengthening of a Giant Protein: When, How, and Why?

Subcommissural organ (SCO)-spondin is a giant glycoprotein of more than 5000 amino acids found in Vertebrata, expressed in the central nervous system and constitutive of Reissner’s fiber. For the first time, in situ hybridization performed on zebrafish (Danio rerio) embryos shows that the gene encoding this protein is expressed transitionally in the floor plate, the ventral midline of the neural tube, and later in the diencephalic third ventricle roof, the SCO. The modular organization of the protein in Echinodermata (Strongylocentrotus purpuratus), Urochordata (Ciona savignyi and C. intestinalis), and Vertebrata (Teleostei, Amphibia, Aves and Mammalia) is also described. As the thrombospondin type 1 repeat motifs represent an increasingly large part of the protein during Deuterostomia evolution, the duplication mechanisms leading to this complex organization are examined. The functional significance of the particularly well-preserved arrangement of the series of SCO-spondin repeat motifs and thombospondin type 1 repeats is discussed. Electronic Supplementary Material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Mapping PrBn and other quantitative trait loci responsible for the control of homeologous chromosome pairing in oilseed rape (Brassica napus L.) haploids

In allopolyploid species, fair meiosis could be challenged by homeologous chromosome pairing and is usually achieved by the action of homeologous pairing suppressor genes. Oilseed rape (Brassica napus) haploids (AC, n=19) represent an attractive model for studying the mechanisms used by allopolyploids to ensure the diploid-like meiotic pairing pattern. In oilseed rape haploids, homeologous chromosome pairing at metaphase I was found to be genetically based and controlled by a major gene, PrBn, segregating in a background of polygenic variation. In this study, we have mapped PrBn within a 10-cM interval on the C genome linkage group DY15 and shown that PrBn displays incomplete penetrance or variable expressivity. We have identified three to six minor QTL/BTL that have slight additive effects on the amount of pairing at metaphase I but do not interact with PrBn. We have also detected a number of other loci that interact epistatically, notably with PrBn. Our results support the idea that, as in other polyploid species, metaphase I homeologous pairing in oilseed rape haploids is controlled by an integrated system of several genes, which function in a complex manner.     

Altered localization of H-Ras in caveolin-1-null cells is palmitoylation-independent

Caveolin-1 is a palmitoylated protein involved in the formation of plasma membrane subdomains termed caveolae, intracellular cholesterol transport, and assembly and regulation of signaling molecules in caveolae. Caveolin-1 interacts via a consensus binding motif with several signaling proteins, including H-Ras. Ras oncogene products function as molecular switches in several signal transduction pathways regulating cell growth and differentiation. Post-translational modifications, including palmitoylation, are critical for the membrane targeting and function of H-Ras. Subcellular localization regulates the signaling pathways engaged by H-Ras activation. We show here that H-Ras is localized at the plasma membrane in caveolin-1-expressing cells but not in caveolin-1-deficient cells. Since palmitoylation is required for trafficking of H-Ras from the endomembrane system to the plasma membrane, we tested whether the altered localization of H-Ras in caveolin-1-null cells is due to decreased H-Ras palmitoylation. Although the palmitoylation profiles of cultured embryo fibroblasts isolated from wild type and caveolin-1 gene-disrupted mice differed, suggesting that caveolin-1, or caveolae, play a role in the palmitate incorporation of a subset of palmitoylated proteins, the palmitoylation of H-Ras was not decreased in caveolin-1-null cells. We conclude that the altered localization of H-Ras in caveolin-1-deficient cells is palmitoylation-independent. This article shows two important new mechanisms by which loss of caveolin-1 expression may perturb intracellular signaling, namely the mislocalization of signaling proteins and alterations in protein palmitoylation.     

Meiotic studies of infertile men in case of non-obstructive azoospermia with normal karyotype and no microdeleted Y-chromosome precise the clinical couple management

To identify meiotic criteria for infertility management in non-obstructive azoospermic men, a prospective and multicentric study was organized in Andrological Departments of Paris (France), Roma (Italy) and Budapest (Hungary). In 117 non-obstructive azoospermic men with normal karyotype and no Y-chromosome microdeletion, histology and meiotic studies on bilateral bipolar testicular biopsies were done. Histologically, 40 patients (34%) presented spermatocyte or spermatid arrest, 39 (33%) hypospermatogenesis whereas no meiotic cell could be observed in the remaining patients (33%). Cytogenetically, meiotic figures could only be obtained from the two first histological groups. Meiotic abnormalities were observed in a total of 44 patients (37.6%) including nine patients (7.7%) with severe class I and class IIB anomalies and 19 patients (16.2%) with class IIC environmentally linked meiotic abnormalities. These results provided essential clues for an accurate clinical management. For patients with no meiotic figures and patients with class I and class IIB anomalies, an hormonal stimulation is illusory and a sperm gift should be directly proposed. An hormonal stimulation should be proposed to all the other patients, either directly or following the treatment of the testicular microenvironment for the patients presenting class IIC anomalies. The genetic risk and possibility of prenatal chromosomal analysis in case of pregnancy should be clearly exposed to all the couples in all the cases where type IIA, III or IV anomalies are present. This therapeutical strategy has been applied to all the patients in our series.     

Neurotrophins are expressed in giant cell arteritis lesions and may contribute to vascular remodeling

Introduction

Giant cell arteritis (GCA) is characterized by intimal hyperplasia leading to ischaemic manifestations that involve large vessels. Neurotrophins (NTs) and their receptors (NTRs) are protein factors for growth, differentiation and survival of neurons. They are also involved in the migration of vascular smooth muscle cells (VSMCs). Our aim was to investigate whether NTs and NTRs are involved in vascular remodelling of GCA.

Methods

We included consecutive patients who underwent a temporal artery biopsy for suspected GCA. We developed an enzymatic digestion method to obtain VSMCs from smooth muscle cells in GCA patients and controls. Neurotrophin protein and gene expression and functional assays were studied from these VSMCs. Neurotrophin expression was also analysed by immunohistochemistry in GCA patients and controls.

Results

Whereas temporal arteries of both GCA patients (n = 22) and controls (n = 21) expressed nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), tropomyosin receptor kinase B (TrkB) and sortilin, immunostaining was more intense in GCA patients, especially in the media and intima, while neurotrophin-3 (NT-3) and P75 receptor (P75^NTR) were only detected in TA from GCA patients. Expression of TrkB, a BDNF receptor, was higher in GCA patients with ischaemic complications. Serum NGF was significantly higher in GCA patients (n = 28) vs. controls (n = 48), whereas no significant difference was found for BDNF and NT-3. NGF and BDNF enhanced GCA-derived temporal artery VSMC proliferation and BDNF facilitated migration of temporal artery VSMCs in patients with GCA compared to controls.

Conclusions

Our results suggest that NTs and NTRs are involved in vascular remodelling of GCA. In GCA-derived temporal artery VSMC, NGF promoted proliferation and BDNF enhanced migration by binding to TrkB and p75^NTR receptors. Further experiments are needed on a larger number of VSMC samples to confirm these results.

Electronic supplementary material

The online version of this article (doi:10.1186/s13075-014-0487-z) contains supplementary material, which is available to authorized users.     

Organization of cytoskeleton during the tentacle contraction and cytostome movement in the dinoflagellate Noctiluca scintillans McCartney

Summary Electron microscopy of Noctiluca scintillans reveals that the cytoskeleton of the tentacle involved in the motor action of the prey capture consists of three characteristic elements: a deformable peripheral fibrillo-granular ectosarc, abundant underlying microtubules organized in several rows on the convex side, and helicoid filaments about 8 nm in diameter organized into striated myonemes. Microtubules of the external row are crossed-linked with each other by fibrous elements 5 nm in diameter and 10–15 nm long, their links with the second row result in a Y-shaped binding. Bonds of the other rows are linked to each other irregularly between those of the same row. Striated myonemes are regularly inserted between the rows of microtubules on the ectosarc and between its pleats, joining together in a knot of disarrayed filaments with multidirectional orientation in the central axis of the tentacle. Striation of myonemes is based on an alternation of thick striae (TS) 40 nm wide with a periodicity of about 200 nm, and of some intermediary fine striae (FS) 10 nm wide. The events during tentacle contraction are: (1) Rotation of the tentacle, bringing the convex side to the inner side of it. Here, large numbers of microtubules have been visualized by optical immunocytochemistry after labelling with Paramecium antitubulin antiserum. (2) Increase of pleat amplitude (200–300 nm to 600 nm) in concomitance with a decrease of its period (500–700 nm to 250 nm). (3) Apparent modification of the microtubule orientation. (4) Transformation of some TS in several FS without modification of the striation periodicity.Near the cytostome, the cytoskeleton consists of a number quantity of microtubules underlying a non-pleated ectosarc and long tracts of contractile myonemes formed by 6-nm helicoid filaments linking the internal side of the cytostome of the supporting rod. Semirelaxed myonemes show an alternation of fine striae (FS) 35 nm wide between two clear areas (CA) with a periodicity of about 300 nm, plus an incipient dark area (DA) lying between them; together they are transformed into a thick stria (TS) during maximal contraction; the striation periodicity thus decreases by one half. These two systems are compared with one another and with other motile systems.     

Effect of Calcium and Calcium-Counteracting Drugs on the Response of Bidens pilosa L. to Wounding

We used calcium counteracting drugs known to reduce the amplitudeof wound-induced electric wave of depolarization and we showedthat in these conditions, accumulation of the calmodulin mRNA(recently found to be correlated to membrane potential) is stronglyreduced. These results bring additional evidence linking membranepotential and calmodulin mRNA accumulation. ⁵Present address: North Carolina State University, Departmentof Botany, BOX 7612, Raleigh, NC 27695, U.S.A.     

Reassessing the role of phosphocaveolin-1 in cell adhesion and migration

Although phosphorylation on tyrosine 14 was identified early in the discovery of caveolin-1, the functional significance of this modification still remains elusive. Recent evidence points to a role of caveolin-1 tyrosine 14 phosphorylation in cell adhesion and migration. These results are based on a variety of tools, including a widely used mouse monoclonal anti-phosphocaveolin-1 antibody, which labels, in cultured cells, a protein localized at or near focal adhesions. We here report results from three independent laboratories, showing that this antibody recognizes phosphocaveolin-1 amongst other proteins in immunoblot analyses and that the signal obtained with this antibody in immunostaining experiments is in part due to labeling of paxillin. Published data need to be interpreted keeping in mind that images of phosphocaveolin-1 cellular localization obtained using this antibody are not valid. We re-evaluate the current knowledge about the role of caveolin-1 in cell adhesion and migration in view of this new information.