Dissection of the transcriptional regulation of grape ASR and response to glucose and abscisic acid

Despite the fact that the precise physiological function of ASRs [abscisic acid (ABA), stress, ripening] remains unknown, they have been suggested to play a dual role in the plant response to environmental cues, as highly hydrophilic proteins for direct protection, as well as transcription factors involved in the regulation of gene expression. To investigate further the biological positioning of grape ASR in the hormonal and metabolic signal network, three promoters corresponding to its cDNA were isolated and submited to a detailed in silico and functional analysis. The results obtained provided evidence for the allelic polymorphism of the grape ASR gene, the organ-preferential expression conferred on the GUS reporter gene, and the specific phloem tissue localization revealed by in situ hybridization. The study of glucose and ABA signalling in its transcriptional control, by transfection of grape protoplasts using the dual luciferase system, revealed the complexity of ASR gene expression regulation. A model was proposed allowing a discussion of the place of ASR in the fine tuning of hormonal and metabolic signalling involved in the integration of environmental cues by the plant organism

Interplay of Sugar, Light and Gibberellins in Expression of Rosa hybrida Vacuolar Invertase 1 Regulation

Our previous findings showed that the expression of the Rosa hybrida vacuolar invertase 1 gene (RhVI1) was tightly correlated with the ability of buds to grow out and was under sugar, gibberellin and light control. Here, we aimed to provide an insight into the mechanistic basis of this regulation. In situ hybridization showed that RhVI1 expression was localized in epidermal cells of young leaves of bursting buds. We then isolated a 895 bp fragment of the promoter of RhVI1. In silico analysis identified putative cis-elements involved in the response to sugars, light and gibberellins on its proximal part (595 bp). To carry out functional analysis of the RhVI1 promoter in a homologous system, we developed a direct method for stable transformation of rose cells. 5′ deletions of the proximal promoter fused to the uidA reporter gene were inserted into the rose cell genome to study the cell’s response to exogenous and endogenous stimuli. Deletion analysis revealed that the 468 bp promoter fragment is sufficient to trigger reporter gene activity in response to light, sugars and gibberellins. This region confers sucrose- and fructose-, but not glucose-, responsive activation in the dark. Inversely, the –595 to –468 bp region that carries the sugar-repressive element (SRE) is required to down-regulate the RhVI1 promoter in response to sucrose and fructose in the dark. We also demonstrate that sugar/light and gibberellin/light act synergistically to up-regulate β-glucuronidase (GUS) activity sharply under the control of the 595 bp pRhVI1 region. These results reveal that the 127 bp promoter fragment located between –595 and –468 bp is critical for light and sugar and light and gibberellins to act synergistically

Efficiency assessment of hybrid coatings for natural building stones : advanced and multi-scale laboratory investigation

The efficiency of a hybrid patented consolidant (PAASi) and two commercially available hybrid coatings (a consolidant named AlSiX and a hydrophobic product named WS3) properly modified was assessed on a calcarenite substrate. Test routines based on standard recommendations were first applied to evaluate the performances of the consolidant and protective treatments, while the investigation of additional aspects such as penetration depth and interaction with the substrate was achieved by a multi-scale approach based on classic intrusion methods (mercury intrusion porosimetry) and Drilling Resistance Measurement System (DRMS), combined with non-invasive imaging techniques (X-ray computed micro-tomography and neutron radiography) and small angle neutron scattering (SANS). A distinct interaction of the products with the pore network of the stone was quantified in the range 0.007–200 µm. Their effects on capillary water absorption were also visualized with neutron imaging. The suitability of the products on the selected substrate was discussed, highlighting also how the applied routine can support conservation material studies. The results indicated that the Al-Si-based product led to unwanted effects. Alternative application methods and/or curing procedures have to be explored to overtake these undesirable changes. On the contrary, the polyamidoamine-based product seemed to be more suitable for calcarenite substrates conservation. The performances of the hydrophobic coating, when used in combination with consolidants, were strictly influenced by the pre-consolidation of the substrate

Multi-scale laboratory routine in the efficacy assessment of conservative products for natural stones

The evaluation of conservative treatments’ efficacy on natural building stones are usually based on standard recommendation routines finalized to evaluate compatibility and harmfulness of products in turn of the substrate. However, the visualization and the quantification of products inside pore structure of natural stones is not immediate through standard tests, so that imaging and advanced techniques are recently proposed in material conservation field to improve knowledge on penetration depth, modification of pore-air interface at different scale and monitor dynamic absorption processes. Moreover, natural stones are usually characterized by complex structure, which changes due to conservative treatments have to be inspected at different scale (from micrometer to nanometer). In this prospective, the assessment of laboratory practices able to integrate multiscale methods and give back a complete overview on interaction between new conservative formulates and natural stones is of high interest. In this paper, we propose a methodological routine for efficacy assessment of conservative products, incorporating classical and innovative nondestructive techniques. Validation of the workflow has been verified on a high porous natural stone treated with new hybrid formulates appropriately customized for conservation issues. • The study intends to add new insights on problems related to consolidation of high porous carbonate stone, application methods in consolidating natural stones and methods to evaluate efficacy of new products.• A multi-scale laboratory investigation procedure is proposed by integrating standard and innovative nondestructive methods. Merits and limits of each applied method are discussed during validation.• The possibility to incorporate standard routines and/or substitute destructive testing with non-destructive ones seem to be a valid alternative to evaluate efficiency and monitor behavior of stones treated with consolidating products

Region-specific sex modulation of central oxytocin receptor by gut microbiota: An ontogenic study.

Oxytocin (OT) is a developmentally important neuropeptide recognized to play a dominant role in social functioning and stress-related behaviors, in a sex-dependent manner. Nonetheless, the underlining factors driving OT and OT receptor (OTR) early brain development remain unclear. Recent evidence highlight the critical influence of gut microbiota and its bidirectional interaction with the brain on neurodevelopment via the gut microbiota-brain axis. Therefore, we aimed to determine the impact of gut microbiota on the OTR system of the rat brain at different developmental stages in a pilot study. Quantitative OTR [125 I]-OVTA autoradiographic binding was carried out in the forebrain of male and female conventional (CON) and germ-free (GF) rats at postnatal days (PND) 8, 22, and 116-150. OTR binding was also assessed in the eyes of PND 1 and PND 4 GF female rats. Significant "microbiota × sex × region" interaction and age-dependent effects on OTR binding were demonstrated. Microbiota status influenced OTR levels in males but not females with higher levels of OTR observed in GF versus CON rats in the cingulate, prelimbic, and lateral/medial/ventral orbital cortex, and septum across all age groups, while sex differences were observed in GF, but not in CON rats. Interestingly, OTRs present in the eyes of CON rats were abolished in GF rats. This is the first study to uncover a sex-specific role of gut microbiota on the central OTR system, which may have implications in understanding the developmental neuroadaptations critical for behavioral regulation and the etiology of certain neurodevelopmental disorders

Impacts of contrasting light on bud burst and on RwMAX1 and RwMAX2 expression in rose

Bud burst is a crucial factor in plant architecture and is strongly induced by light. In Rosa sp., this light effect was correlated with the growth of axillary buds and RwMAX1 and RwMAX2 expression within buds. In this paper, we investigated whether strigolactone pathway is involved in the regulation of axillary bud in response to light intensity. Hence, young roses were subjected to two contrasting light intensity regimes: high/high and high/low. The phenotype was characterized in both conditions and the expression of RwMAX1 and RwMAX2 genes was measured in the basal, middle and apical parts of rose primary branch. Light treatments showed a strong impact on axillary bud. The percentage of bud burst was severely reduced in the treatment high/low compared to the treatment high/high in all branch parts. In addition, the expression of RwMAX1 and RwMAX2 was strongly inhibited by high/high light regime and was conversely correlated with the rate of bud burst. In in vitro-grown axillary buds supplied with sucrose, glucose and fructose, RwMAX1 expression was significantly stimulated whereas that of RwMAX2 was significantly inhibited. Our results suggest that although RwMAX1 and RwMAX2 expression can be regulated by light, this expression does not explain the ability of bud burst

A Conserved Mito-Cytosolic Translational Balance Links Two Longevity Pathways.

Slowing down translation in either the cytosol or the mitochondria is a conserved longevity mechanism. Here, we found a non-interventional natural correlation of mitochondrial and cytosolic ribosomal proteins (RPs) in mouse population genetics, suggesting a translational balance. Inhibiting mitochondrial translation in C. elegans through mrps-5 RNAi repressed cytosolic translation. Transcriptomics integrated with proteomics revealed that this inhibition specifically reduced translational efficiency of mRNAs required in growth pathways while increasing stress response mRNAs. The repression of cytosolic translation and extension of lifespan from mrps-5 RNAi were dependent on atf-5/ATF4 and independent from metabolic phenotypes. We found the translational balance to be conserved in mammalian cells upon inhibiting mitochondrial translation pharmacologically with doxycycline. Lastly, extending this in vivo, doxycycline repressed cytosolic translation in the livers of germ-free mice. These data demonstrate that inhibiting mitochondrial translation initiates an atf-5/ATF4-dependent cascade leading to coordinated repression of cytosolic translation, which could be targeted to promote longevity

SEX-DEPENDENT IMPACT OF MICROBIOTA STATUS ON CEREBRAL μ -OPIOID RECEPTOR DENSITY IN FISCHER RATS.

μ-opioid receptors (MOPr) play a critical role in social play, reward, and pain, in a sex and age-dependent manner. There is evidence to suggest that sex and age differences in brain MOPr density may be responsible for this variability, however, little is known about the factors driving these differences in cerebral MOPr density. Emerging evidence highlights gut microbiota's critical influence and its bidirectional interaction with the brain on neurodevelopment. Therefore, we aimed to determine the impact of gut microbiota on MOPr density in male and female brains at different developmental stages. Quantitative [3 H]DAMGO autoradiographic binding was carried out in the forebrain of male and female conventional (CON), and germ-free (GF) rats at postnatal days (PND) 8, 22, and 116-150. Significant 'microbiota status x sex,' 'age x brain region' interactions, and microbiota status- and age-dependent effects on MOPr binding were uncovered. Microbiota status influenced MOPr levels in males but not females, with higher MOPr levels observed in GF vs. CON rats overall regions and age groups. In contrast, no overall sex differences were observed in GF or CON rats. Interestingly, within-age planned comparison analysis conducted in frontal cortical and brain regions associated with reward revealed that this microbiota effect was restricted only to PND22 rats. Thus, this pilot study uncovers the critical sex-dependent role of gut microbiota in regulating cerebral MOPr density, which is restricted to the sensitive developmental period of weaning. This may have implications in understanding the importance of microbiota during early development on opioid signalling and associated behaviours

Insight into the Role of Sugars in Bud Burst Under Light in the Rose

Bud burst is a decisive process in plant architecture that requires light in Rosa sp. This light effect was correlated with stimulation of sugar transport and metabolism in favor of bud outgrowth. We investigated whether sugars could act as signaling entities in the light-mediated regulation of vacuolar invertases and bud burst. Full-length cDNAs encoding two vacuolar invertases (RhVI1 and RhVI2) were isolated from buds. Unlike RhVI2, RhVI1 was preferentially expressed in bursting buds, and was up-regulated in buds of beheaded plants exposed to light. To assess the importance of sugars in this process, the expression of RhVI1 and RhVI2 and the total vacuolar invertase activity were further characterized in buds cultured in vitro on 100 mM sucrose or mannitol under light or in darkness for 48 h. Unlike mannitol, sucrose promoted the stimulatory effect of light on both RhVI1 expression and vacuolar invertase activity. This up-regulation of RhVI1 was rapid (after 6 h incubation) and was induced by as little as 10 mM sucrose or fructose. No effect of glucose was found. Interestingly, both 30 mM palatinose (a non-metabolizable sucrose analog) and 5 mM psicose (a non-metabolizable fructose analog) promoted the light-induced expression of RhVI1 and total vacuolar invertase activity. Sucrose, fructose, palatinose and psicose all promoted bursting of in vitro cultured buds under light. These findings indicate that soluble sugars contribute to the light effect on bud burst and vacuolar invertases, and can function as signaling entities