Phenotypic switch of smooth muscle cells in paediatric chronic intestinal pseudo-obstruction syndrome

Smooth Muscle Cells (SMC) are unique amongst all muscle cells in their capacity to modulate their phenotype. Indeed, SMCs do not terminally differentiate but instead harbour a remarkable capacity to dedifferentiate, switching between a quiescent contractile state and a highly proliferative and migratory phenotype, a quality often associated to SMC dysfunction. However, phenotypic plasticity remains poorly examined in the field of gastroenterology in particular in pathologies in which gut motor activity is impaired. Here, we assessed SMC status in biopsies of infants with chronic intestinal pseudo-obstruction (CIPO) syndrome, a life-threatening intestinal motility disorder. We showed that CIPO-SMCs harbour a decreased level of contractile markers. This phenotype is accompanied by an increase in Platelet-Derived Growth Factor Receptor-alpha (PDGFRA) expression. We showed that this modulation occurs without origin-related differences in CIPO circular and longitudinal-derived SMCs. As we characterized PDGFRA as a marker of digestive mesenchymal progenitors during embryogenesis, our results suggest a phenotypic switch of the CIPO-SMC towards an undifferentiated stage. The development of CIPO-SMC culture and the characterization of SMC phenotypic switch should enable us to design therapeutic approaches to promote SMC differentiation in CIPO.     

Hyperexpression of recombinant CFTR in heterologous cells alters its physiological properties

We investigated whether high levels of expression of the cysticfibrosis transmembrane conductance regulator (CFTR) would alter thefunctional properties of newly synthesized recombinant proteins. COS-7,CFPAC-1, and A549 cells were intranuclearly injected with a Simianvirus 40-driven pECE-CFTR plasmid and assayed for halide permeabilityusing the6-methoxy-N-(3-sulfopropyl)quinolinium fluorescent probe. With increasing numbers of microinjected pECE-CFTR copies, the baseline permeability to halide dose dependently increased, and the response to adenosine 3',5'-cyclic monophosphate(cAMP) stimulation decreased. In cells hyperexpressing CFTR, the high level of halide permeability was reduced when a cell metabolism poisoning cocktail was applied to decrease intracellular ATP and, inversely, was increased by orthovanadate. In CFPAC-1 cellsinvestigated with the patch-clamp technique, CFTR hyperexpression ledto a time-independent nonrectifying chloride current that was notsensitive to cAMP stimulation. CFPAC-1 cells hyperexpressing CFTRexhibited no outward rectifying chloride current nor inward rectifyingpotassium current either spontaneously or under cAMP stimulation. Weconclude that hyperexpression of recombinant CFTR proteins modifiestheir properties inasmuch as 1) CFTRchannels are permanently activated and not susceptible to cAMPregulation and 2) they lose their capacity to regulate heterologous ionic channels.

     

Sexual Dimorphism of the Feto-Placental Phenotype in Response to a High Fat and Control Maternal Diets in a Rabbit Model

Maternal environment during early developmental stages plays a seminal role in the establishment of adult phenotype. Using a rabbit model, we previously showed that feeding dams with a diet supplemented with 8% fat and 0.2% cholesterol (HH diet) from the prepubertal period and throughout gestation induced metabolic syndrome in adult offspring. Here, we examined the effects of the HH diet on feto-placental phenotype at 28 days post-coïtum (term = 31days) in relation to earlier effects in the blastocyst (Day 6). At 28 days, both male and female HH fetuses were intrauterine growth retarded and dyslipidemic, with males more affected than females. Lipid droplets accumulated in the HH placentas’ trophoblast, consistent with the increased concentrations in cholesteryl esters (3.2-fold), triacylglycerol (2.5-fold) and stored FA (2.12-fold). Stored FA concentrations were significantly higher in female compared to male HH placentas (2.18-fold, p<0.01), whereas triacylglycerol was increased only in HH males. Trophoblastic lipid droplet accumulation was also observed at the blastocyst stage. The expression of numerous genes involved in lipid pathways differed significantly according to diet both in term placenta and at the blastocyst stage. Among them, the expression of LXR-α in HH placentas was reduced in HH males but not females. These data demonstrate that maternal HH diet affects the blastocyst and induces sex-dependent metabolic adaptations in the placenta, which appears to protect female fetuses from developing severe dyslipidemia.     

Queuosine Biosynthesis Is Required for Sinorhizobium meliloti-Induced Cytoskeletal Modifications on HeLa Cells and Symbiosis with Medicago truncatula

Rhizobia are symbiotic soil bacteria able to intracellularly colonize legume nodule cells and form nitrogen-fixing symbiosomes therein. How the plant cell cytoskeleton reorganizes in response to rhizobium colonization has remained poorly understood especially because of the lack of an in vitro infection assay. Here, we report on the use of the heterologous HeLa cell model to experimentally tackle this question. We observed that the model rhizobium Sinorhizobium meliloti, and other rhizobia as well, were able to trigger a major reorganization of actin cytoskeleton of cultured HeLa cells in vitro. Cell deformation was associated with an inhibition of the three major small RhoGTPases Cdc42, RhoA and Rac1. Bacterial entry, cytoskeleton rearrangements and modulation of RhoGTPase activity required an intact S. meliloti biosynthetic pathway for queuosine, a hypermodifed nucleoside regulating protein translation through tRNA, and possibly mRNA, modification. We showed that an intact bacterial queuosine biosynthetic pathway was also required for effective nitrogen-fixing symbiosis of S. meliloti with its host plant Medicago truncatula, thus indicating that one or several key symbiotic functions of S. meliloti are under queuosine control. We discuss whether the symbiotic defect of que mutants may originate, at least in part, from an altered capacity to modify plant cell actin cytoskeleton.     

Mobility affects copulation and oviposition dynamics in Pieris brassicae in seminatural cages

When, how often and for how long organisms mate can have strong consequences for individual fitness and are crucial aspects of evolutionary ecology. Such determinants are likely to be of even greater importance in monandrous species and species with short adult life stages. Previous work suggests that mobility, a key dispersal? related trait, may affect the dynamics of copulations, but few studies have investigated the impact of individual mobility on mating latency, copulation duration and oviposition latency simultaneously. In this paper, we monitored the copulation dynamics of 40 males and 40 females, as well as the oviposition dynamics of the females of the Large White butterfly Pieris brassicae, a facultative long-distance disperser butterfly. Individuals from a breeding were selected to create a uniform distribution of mobility and we recorded the timing, number and duration of all copulations in a semiexperimental system. We showed that mobility, measured as the time spent in flight under stressful conditions (a proxy of dispersal tendency), correlates with all aspects of copulation dynamics: mobile males and females mated earlier and for shorter periods than less mobile individuals. In turn, late mating females increased the time between copulation and oviposition. These results feed the previously described mobility syndrome of R brassicae, involving morphological and physiological characters, with life-history traits. We suggest that the reduction of mating latency and copulation duration has an adaptive value in dispersing individuals, as their life expectancy might be shorter than that of sedentary individuals.     

Precise Gene Editing Preserves Hematopoietic Stem Cell Function following Transient p53-Mediated DNA Damage Response

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