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91.
Coïc YM Baleux F Poyraz Ö Thibeaux R Labruyere E Chretien F Sobhani I Lazure T Wyplosz B Schneider G Mulard L Sansonetti PJ Marteyn BS 《The Journal of biological chemistry》2012,287(19):15916-15922
Imaging living cells and organs requires innovative, specific, efficient, and well tolerated fluorescent markers targeting cellular components. Such tools will allow proceeding to the dynamic analysis of cells and the adaptation of tissues to environmental cues. In this study, we have identified and synthesized a novel non-toxic fluorescent marker allowing a specific fluorescent staining of the human colonic mucus. Our strategy to identify a molecule able to specifically bind to the human colonic mucus was on the basis of the mucus adhesion properties of commensal bacteria. We identified and characterized the mucus-binding property of a 70-amino acid domain (MUB(70)) expressed on the surface of Lactobacillus strains. The chemical synthesis of MUB(70) was achieved using the human commensal bacterium Lactobacillus reuteri AF120104 protein as a template. The synthesized Cy5-conjugated MUB(70) marker specifically stained the colonic mucus on fixed human, rabbit, and guinea pig tissues. Interestingly, murine tissue was not stained, suggesting significant differences in the composition of the murine colonic mucus. In addition, this marker stained the mucus of living cultured human colonic cells (HT29-MTX) and human colonic tissue explants. Using a biotinylated derivative of MUB(70), we demonstrated that this peptide binds specifically to Muc2, the most abundant secreted mucin, through its glycosylated moieties. Hence, Cy5-MUB(70) is a novel and specific fluorescent marker for mammalian colonic mucus. It may be used for live imaging analysis but also, as demonstrated in this study, as a marker for the diagnosis and the prognosis of colonic mucinous carcinomas. 相似文献
92.
Perroy J Raynaud F Homburger V Rousset MC Telley L Bockaert J Fagni L 《The Journal of biological chemistry》2008,283(11):6799-6805
Functional interplay between ionotropic and metabotropic receptors frequently involves complex intracellular signaling cascades. The group I metabotropic glutamate receptor mGlu5a co-clusters with the ionotropic N-methyl-d-aspartate (NMDA) receptor in hippocampal neurons. In this study, we report that a more direct cross-talk can exist between these types of receptors. Using bioluminescence resonance energy transfer in living HEK293 cells, we demonstrate that mGlu5a and NMDA receptor clustering reflects the existence of direct physical interactions. Consequently, the mGlu5a receptor decreased NMDA receptor current, and reciprocally, the NMDA receptor strongly reduced the ability of the mGlu5a receptor to release intracellular calcium. We show that deletion of the C terminus of the mGlu5a receptor abolished both its interaction with the NMDA receptor and reciprocal inhibition of the receptors. This direct functional interaction implies a higher degree of target-effector specificity, timing, and subcellular localization of signaling than could ever be predicted with complex signaling pathways. 相似文献
93.
Prunier F Marescaux L Franconi F Thia A Legras P Lejeune JJ Furber A 《Canadian journal of physiology and pharmacology》2005,83(12):1109-1115
In vivo assessment of treatment efficacy on postinfarct left ventricular (LV) remodeling is crucial for experimental studies. We examined the technical feasibility of serial magnetic resonance imaging (MRI) for monitoring early postinfarct remodeling in rats. MRI studies were performed with a 7-Tesla unit, 1, 3, 8, 15, and 30 days after myocardial infarction (MI) or sham operation, to measure LV mass, volume, and the ejection fraction (EF). Three groups of animals were analyzed: sham-operated rats (n = 6), MI rats receiving lisinopril (n = 11), and MI rats receiving placebo (n = 8). LV dilation occurred on day 3 in both MI groups. LV end-systolic and end-diastolic volumes were significantly lower in lisinopril-treated rats than in placebo-treated rats at days 15 and 30. EF was lower in both MI groups than in the sham group at all time points, and did not differ between the MI groups during follow-up. Less LV hypertrophy was observed in rats receiving lisinopril than in rats receiving placebo at days 15 and 30. We found acceptable within- and between-observer agreement and an excellent correlation between MRI and ex vivo LV mass (r = 0.96; p < 0.001). We demonstrated the ability of MRI to detect the early beneficial impact of angiotensin-converting enzyme (ACE) inhibitors on LV remodeling. Accurate and noninvasive, MRI is the tool of choice to document response to treatment targeting postinfarction LV remodeling in rats. 相似文献
94.
Gosset P Bureau F Angeli V Pichavant M Faveeuw C Tonnel AB Trottein F 《Journal of immunology (Baltimore, Md. : 1950)》2003,170(10):4943-4952
Among the factors produced at inflammatory sites and those capable of modulating dendritic cell (DC) functions, PGD(2) may be important in the outcome of immune responses. The biological roles for PGD(2) are in part effected through two plasma membrane G protein-coupled receptors: the D prostanoid (DP) receptor and the chemoattractant receptor-homologous molecule expressed on Th2 lymphocytes (CRTH2). In this report, we studied the effects of PGD(2) and of its major physiological metabolite, 15-deoxy-Delta(12,14)-PGJ(2) (15d-PGJ(2)), on the functions of human monocyte-derived DC. First, we show that PGD(2) exerts in vitro chemotactic effects on monocytes via CRTH2 activation while it inhibits the chemokine-driven migration of monocyte-derived DC through DP. We also report that PGD(2) and 15d-PGJ(2) alter the LPS- and allergen-induced DC maturation and enhance the CD80/CD86 ratio on mature DC in a DP- and CRTH2-independent manner. Moreover, PGD(2) and 15d-PGJ(2) strongly reduce the secretion of the Th1 promoting cytokine IL-12 and affect the synthesis of chemokines involved in Th1 cell chemotaxis, particularly CXCL10. Inhibition of cytokine/chemokine secretion implicates at least in part DP, but not CRTH2. The effects exerted by PGD(2) are associated with the phosphorylation of CREB, but do not parallel with the deactivation of the NF-kappa B and mitogen-activated protein kinase pathways. In contrast, 15d-PGJ(2) seems to target other cellular proteins. Finally, in a model of Th CD45RA(+) differentiation induced by allergen- and superantigen-pulsed DC, PGD(2) impacts on the orientation of the immune response by favoring a Th2 response. 相似文献
95.
Vasectomy has been shown to affect the pattern of mRNA expression of P34H, a human sperm protein added to the acrosomal cap during epididymal transit. It has been reported that vasectomy alters the histology of the reproductive tract in various species as a result of the increased pressure in the epididymis. The aim of this study was to evaluate if other epididymis-specific mRNAs, which are expressed in different patterns along the duct, are altered by vasectomy as well. We analyzed the expression of P31m (a monkey homologue of human P34H) and three different HE-like (HE-l) mRNAs along the epididymis in the cynomolgus monkey (Macaca fascicularis). Sexually mature cynomolgus monkeys were vasectomized unilaterally; then the epididymides were surgically removed at different time points. The ipsilateral normal epididymis was used as a control. Histomorphometric measurements showed that the height of the epididymal epithelial cells started to be affected only at 14 wk postsurgery. However, Northern blot and in situ hybridization analysis showed that the expression pattern of P31m, HE1, and HE5-like mRNA along the epididymis was not affected by vasectomy. Only the HE2-like mRNA predominantly expressed in the normal corpus epididymidis was significantly lowered 14 wk after vasectomy. Thus, ductal obstruction differentially alters mRNA expression along the epididymis of the cynomolgus monkey. 相似文献
96.
Rappaport Fabrice Béal Daniel Verméglio André Joliot Pierre 《Photosynthesis research》1998,55(2-3):317-323
We have studied the electron transfer reactions from the tetraheme cytochrome of Rhodopseudomonas viridis to the oxidized primary donor in whole cells with a new high sensitivity spectrophotometer. In this apparatus the monochromatic detecting flashes are provided by a YAG pumped Optical Parametric Oscillator, allowing a 10 ns time resolution. When four hemes are reduced the observed electron transfer reaction sequence is the following: first the low-potential c552 heme (the number refers to the maximum absorption wavelength in the alpha-band region) is oxidized with a half time of 130 ns, in agreement with previous reports of measurements performed with purified reaction centers. Then, the electron hole is transferred to the low potential c554 heme with a half time of 2.6 µs. When only the two high potential hemes are reduced the observed electron transfer sequence is the following: oxidation of the high potential c559 heme in the hundreds of ns time range (410 ns), reduction of this heme by the high potential c556 heme in the µs time range (2.7 µs). This confirms the first steps of electron transfer observed in isolated reaction centers. However, in the microsecond time domain, the overall amount of oxidized hemes increases suggesting that, in vivo, the equilibrium constant between the P+/P and the c559ox/c559red couples is significantly lower than expected from the difference in their midpoint potentials. 相似文献
97.
Caudron F Denarier E Thibout-Quintana JC Brocard J Andrieux A Fourest-Lieuvin A 《PloS one》2010,5(10):e13553
Ser172 of β tubulin is an important residue that is mutated in a human brain disease and phosphorylated by the cyclin-dependent kinase Cdk1 in mammalian cells. To examine the role of this residue, we used the yeast S. cerevisiae as a model and produced two different mutations (S172A and S172E) of the conserved Ser172 in the yeast β tubulin Tub2p. The two mutants showed impaired cell growth on benomyl-containing medium and at cold temperatures, altered microtubule (MT) dynamics, and altered nucleus positioning and segregation. When cytoplasmic MT effectors Dyn1p or Kar9p were deleted in S172A and S172E mutants, cells were viable but presented increased ploidy. Furthermore, the two β tubulin mutations exhibited synthetic lethal interactions with Bik1p, Bim1p or Kar3p, which are effectors of cytoplasmic and spindle MTs. In the absence of Mad2p-dependent spindle checkpoint, both mutations are deleterious. These findings show the importance of Ser172 for the correct function of both cytoplasmic and spindle MTs and for normal cell division. 相似文献
98.
The pulsing of river discharge affects biodiversity and productivity of whole river–floodplain ecosystems, triggering the transport, storage and processing of carbon. In this study we investigate the short-term changes in water chemistry and net pelagic metabolism (NEP) in two floodplain lakes in response to a flood pulse. The two oxbow lakes investigated in the floodplain of the Mediterranean Ebro River (NE Spain) showed a clear temporal shift in their metabolic balance, controlled by the river discharge and associated changes in water physical and chemical characteristics. Water chemistry (turbidity, water organic matter, chlorophyll a and nutrients concentration) returned to pre-flood values after 4 days, highlighting the resilience of the ecosystem to flood pulses. Lake NEP was depressed before and during floods to a minimum of −34 mg O2 m−3 h−1, and increased after the flood pulse to a maximum of +463 mg O2 m−3 h−1. The phytoplankton assemblage showed before and after floods a replacement of autotrophic species (e.g. Chlorophyceans) by mixotrophic organisms (e.g. Cryptophyceans, Euglenoids). A linear mixed effects model identified abiotic factors, particularly temperature and river discharge, as significant predictors of the net aquatic metabolism and community respiration during flood conditions. Our results suggest that the role of the Ebro floodplain lakes as sources or sinks of C is complex and relative to the time scale investigated, depending strongly on the river discharge dynamics and the transport of limiting nutrients (phosphorus). 相似文献
99.
Laurent J. Catoire Manuela Zoonens Carine van Heijenoort Fabrice Giusti Éric Guittet Jean-Luc Popot 《European biophysics journal : EBJ》2010,39(4):623-630
The atomic structure of OmpX, the smallest member of the bacterial outer membrane protein family, has been previously established
by X-ray crystallography and NMR spectroscopy. In apparent conflict with electrophysiological studies, the lumen of its transmembrane
β-barrel appears too tightly packed with amino acid side chains to let any solute flow through. In the present study, high-resolution
solution NMR spectra were obtained of OmpX kept water-soluble by either amphipol A8-35 or the detergent dihexanoylphosphatidylcholine.
Hydrogen/deuterium exchange measurements performed after prolonged equilibration show that, whatever the surfactant used,
some of the amide protons of the membrane-spanning region exchange much more readily than others, which likely reflects the
dynamics of the barrel. 相似文献
100.
Fanny Jaudon Fabrice Raynaud Rosine Wehrlé Jean-Michel Bellanger Mohamed Doulazmi Guilan Vodjdani Stéphane Gasman Laurent Fagni Isabelle Dusart Anne Debant Susanne Schmidt 《Molecular biology of the cell》2015,26(11):2112-2127
By regulating actin cytoskeleton dynamics, Rho GTPases and their activators RhoGEFs are implicated in various aspects of neuronal differentiation, including dendritogenesis and synaptogenesis. Purkinje cells (PCs) of the cerebellum, by developing spectacular dendrites covered with spines, represent an attractive model system in which to decipher the molecular signaling underlying these processes. To identify novel regulators of dendritic spine morphogenesis among members of the poorly characterized DOCK family of RhoGEFs, we performed gene expression profiling of fluorescence-activated cell sorting (FACS)-purified murine PCs at various stages of their postnatal differentiation. We found a strong increase in the expression of the Cdc42-specific GEF DOCK10. Depleting DOCK10 in organotypic cerebellar cultures resulted in dramatic dendritic spine defects in PCs. Accordingly, in mouse hippocampal neurons, depletion of DOCK10 or expression of a DOCK10 GEF-dead mutant led to a strong decrease in spine density and size. Conversely, overexpression of DOCK10 led to increased spine formation. We show that DOCK10 function in spinogenesis is mediated mainly by Cdc42 and its downstream effectors N-WASP and PAK3, although DOCK10 is also able to activate Rac1. Our global approach thus identifies an unprecedented function for DOCK10 as a novel regulator of dendritic spine morphogenesis via a Cdc42-mediated pathway. 相似文献