Studying the role of the strawberry Fra protein family in the flavonoid metabolism during fruit ripening

Strawberry fruits are highly appreciated worldwide due to their pleasant flavor and aroma and to the health benefits associated to their consumption. An important part of these properties is due to their content in secondary metabolites, especially phenolic compounds, of which flavonoids are the most abundant in the strawberry fruit. Although the flavonoid biosynthesis pathway is uncovered, little is known about its regulation. The strawberry Fra a (Fra) genes constitute a large family of homologs of the major birch pollen allergen Bet v 1 and for which no equivalents exist in Arabidopsis. Our group has shown that Fra proteins are involved in the formation of colored compounds in strawberries (Muñoz et al., 2010), which mainly depends on the production of certain flavonoids; that they are structurally homologs to the PYR/PYL/RCAR Arabidopsis ABA receptor, and that they are able to bind flavonoids (Casañal et al., 2013). With these previous results, our working hypothesis is that the Fra proteins are involved in the regulation of the flavonoids pathway. They would mechanistically act as the ABA receptor, binding a protein interactor and a ligand to regulate a signaling cascade and/or act as molecular carriers. The main objective of this research is to characterize the Fra family in strawberry and gain insight into their role in the flavonoid metabolism. By RNAseq expression analysis in ripening fruits we have identified transcripts for 10 members of the Fra family. Although expressed in all tissues analyzed, each family member presents a unique pattern of expression, which suggests functional specialization for each Fra protein. Then, our next approach was to identify the proteins that interact with Fras and their ligands to gain knowledge on the role that these proteins play in the flavonoids pathway. To identify the interacting partners of Fras we have performed a yeast two hybrid (Y2H) screening against cDNA libraries of strawberry fruits at the green and red stages. A protein that shares a 95% homology to the Heat stress transcription factor A-4-C like of Fragaria vesca (HSA4C) interacts specifically with Fra1 and not with other family members, which suggests functional diversification of Fra proteins in specific signaling pathways. The Y2H screening is not yet saturated, so characterization of other interacting proteins with other members of the Fra family will shed light on the functional diversity within this gene family. This research will contribute to gain knowledge on how the flavonoid pathway, and hence, the fruit ripening, is regulated in strawberry; an economically important crop but for which basic research is still very limited. References: Muñoz, C, et al. (2010). The Strawberry Fruit Fra a Allergen Functions in Flavonoid Biosynthesis. Molecular Plant, 3(1): 113–124. Casañal, A, et al (2013). The Strawberry Pathogenesis-related 10 (PR-10) Fra a Proteins Control Flavonoid Biosynthesis by Binding Metabolic Intermediates. Journal of Biological Chemistry, 288(49): 35322–35332.

Potential protective role of reactive astrocytes in the periventricular parenchyma in congenital hydrocephalus

Background Cerebrospinal fluid accumulation in hydrocephalus produces an elevation of intraventricular pressure with pathological consequences on the periventricular brain parenchyma including ischemia, oedema, oxidative stress, and accumulation of metabolic waste products. Here we studied in the hyh mouse, an animal model of congenital hydrocephalus, the role of reactive astrocytes in this clinical degenerative condition. Materials and Methods Wild type and hydrocephalic hyh mice at 30 days of postnatal age were used. Three metabolites related to the oxidative and neurotoxic conditions were analysed in ex vivo samples (glutathione, glutamine and taurine) using High Resolution Magic Angle Spinning (HR-MAS). Glutathione synthetase and peroxidase, glutamine synthetase, kidney-type glutaminase (KGA), and taurine/taurine transporter were immunolocated in brain sections. Results Levels of the metabolites were remarkably higher in hydrocephalic conditions. Glutathione peroxidase and synthetase were both detected in the periventricular reactive astrocytes and neurons. Taurine was mostly found free in the periventricular parenchyma and in the reactive astrocytes, and the taurine transporter was mainly present in the neurons located in such regions. Glutamine synthetase was found in reactive astrocytes. Glutaminase was also detected in the reactive astrocytes and in periventricular neurons. These results suggest a possible protective response of reactive astrocytes against oxidative stress and neurotoxic conditions. Conclusions Astrocyte reaction seems to trigger an anti-oxidative and anti-neurotoxic response in order to ameliorate pathological damage in periventricular areas of the hydrocephalic mice.

Moderating effects of gender in the relationships between daily stress, coping styles and hopelessness depression in adolescents

The hopelessness depression (HD) theory assumes a number of contributing factors to the development of hopelessness expectations and HD symptoms. Studies carried out to test HD theory have generally supported the prediction that a negative attributional style contributes to the development of hopelessness expectations and HD symptoms, although the evidence is less consistent for adolescents. Due to these inconsistent results, as well as the assumption advanced by HD theory that other factors aside from attributional style may contribute to the development of HD, in this study we examined the potential role of adolescents’ daily stress and coping styles as contributing factors to HD at these ages, and the effects of gender on these relationships. Seiffge-Krenke (1995) showed that stress in daily life plays a particularly relevant role during adolescence and described three coping styles used by adolescents to face daily stress: active and internal approach-oriented styles, which are considered functional and complementary, and avoidant style, considered dysfunctional. In this study, secondary students (N = 480; aged 13–17) completed the Hopelessness Scale (Beck, Weissman, Lester, & Trexler, 1974), the Hopelessness Depression Symptoms Questionnaire (Metalsky & Joiner, 1997), the Problem Questionnaire ((Seiffge-Krenke, 1995) and the Coping Across Situations Questionnaire (Seiffge-Krenke, 1995). To test the role of daily stress and coping styles in the prediction of hopelessness expectations and HD symptoms, two four-step hierarchical multiple regression analyses were conducted. In these analyses, gender was introduced first, daily stress was introduced second, the three coping styles were entered in a third step, and the moderating effects of gender on the associations of the predictor variables (daily stress and each coping style) with the criterion variables (hopelessness expectations or HD symptoms) were introduced in a fourth step. Results showed a moderating effect of gender on the relationship between daily stress and hopelessness expectations, which revealed a significant effect for boys despite the fact that girls experienced more daily stress than boys, thus suggesting an inoculation effect in girls. It was also found a gender-dependent role of coping styles in the prediction of HD symptoms, revealing a protective effect for the active style in girls and for the internal style in boys. This suggests that girls would benefit from being more action-prone and boys more reflection-prone in order to prevent HD. These findings indicate that programmes aimed to promote coping skills in adolescents would benefit from being gender-adapted.