Funktionale Analyse nicht-konservierter Domänen der Lef/Tcf Transkriptionsfaktoren aus Xenopus laevis

Die Lef/Tcf Transkriptionsfaktoren sind die nuklearen Mediatoren des kanonischen Wnt Signalwegs. Diese Arbeit zeigt auf, dass während der Vertebraten Entwicklung entweder jedes Lef/Tcf ursprüngliche Eigenschaften verloren oder Pangolin (Drosophila dTcf) seit dem Abspalten vom gemeinsamen Vorläufer neue Eigenschaften angenommen hat

Regulation of distinct branches of the non-canonical Wnt-signaling network in Xenopus dorsal marginal zone explants

Background: A tight regulation of the Wnt-signaling network, activated by 19 Wnt molecules and numerous receptors and co-receptors, is required for the establishment of a complex organism. Different branches of this Wnt-signaling network, including the canonical Wnt/β-catenin and the non-canonical Wnt/PCP, Wnt/Ror2 and Wnt/Ca$^{2+}$ pathways, are assigned to distinct developmental processes and are triggered by certain ligand/receptor complexes. The Wnt-signaling molecules are closely related and it is still on debate whether the information for activating a specific branch is encoded by specific sequence motifs within a particular Wnt protein. The model organism Xenopus offers tools to distinguish between Wnt-signaling molecules activating distinct branches of the network. Results: We created chimeric Wnt8a/Wnt11 molecules and could demonstrate that the C-terminal part (containing the BS2) of Wnt8a is responsible for secondary axis formation. Chimeric Wnt11/Wnt5a molecules revealed that the N-terminus with the elements PS3-1 and PS3-2 defines Wnt11 specificity, while elements PS3-1, PS3-2 and PS3-3 are required for Wnt5a specificity. Furthermore, we used Xenopus dorsal marginal zone explants to identify non-canonical Wnt target genes regulated by the Wnt5a branch and the Wnt11 branch. We found that pbk was specifically regulated by Wnt5a and rab11fip5 by Wnt11. Overexpression of these target genes phenocopied the overexpression of their regulators, confirming the distinct roles of Wnt11 and Wnt5a triggered signaling pathways. Furthermore, knock-down of pbk was able to restore convergent extension movements in Wnt5a morphants. Conclusions: The N-terminal part of non-canonical Wnt proteins decides whether the Wnt5a or the Wnt11 branch of the Wnt-signaling network gets activated. The different non-canonical Wnt branches not only regulate cellular behavior, but, surprisingly, also regulate the expression of different target genes. One of these target genes, pbk, seems to be the relevant target gene executing Wnt5a-mediated regulation of convergent extension movements

Anemonopsis macrophylla Sieb. et Zucc.

原著和名: レンゲショウマ科名: キンポウゲ科 = Ranunculaceae採集地: 神奈川県 丹沢山 (相模 丹沢山)採集日: 1962/8/31採集者: 萩庭丈壽整理番号: JH041942国立科学博物館整理番号: TNS-VS-99194

Additional file 4: Figure S3. of Regulation of distinct branches of the non-canonical Wnt-signaling network in Xenopus dorsal marginal zone explants

All chimeras are translated into a protein of the expected size biologically active. In vitro transcribed and translated biotinylated proteins of the chimeric constructs were detected on a western blot via an AP conjugated Streptavidin antibody and visualized with NBT/BCIP. (A) The chimeras between the canonical xWnt8a and the non-canonical xWnt11 are translated in a protein of the expected size. (B) All non-canonical Wnt chimeras are translated in a protein of the expected size. (C) ATF2-luciferase reporter assay of HEK293 cells. All non-canonical chimera pairs are biologically active. Shown is the fold activation of the non-canonical ATF2-luciferase reporter of two independent sets of experiments. The differences in activation between the two sets of experiments are due to different batches of HEK293 cells. In both sets of experiments the chimeric constructs activate the ATF-luciferase reporter in a similar manner as wild-type Wnts. Thus, the chimeras are biologically active non-canonical Wnts. N: number of biological replicates, n: number of independent transfections; * P < 0.05, ** P < 0.01, *** P < 0.001 according to Student’s t test. (TIF 676 kb

Additional file 8: Figure S6. of Regulation of distinct branches of the non-canonical Wnt-signaling network in Xenopus dorsal marginal zone explants

Dynamic expression of pbk and rab11fip5. Maternally expressed pbk mRNA (A) is localized at the animal half of the embryo. At gastrula stages no enrichment of pbk transcripts at distinct regions is visible. However, half-dissected embryos revealed pbk expression mainly in the mesoderm. During the neurula stage, pbk mRNA is localized mainly in the developing CNS including the eyes. This localization persists in the tailbud stage. Transversal sections (1 and 2) indicate enriched pbk mRNA in the dorsal part of the neural tube (arrows). Until gastrula stages rab11fip5 expression is similar to pbk expression: enriched in the animal half and later concentrated in the mesoderm. From the early neurula stage onward rab11fip5 is enriched the anterior neuroectoderm (sections 1 and 2). From the late neurula stage onward, an additional ring shaped expression domain is found around the cement gland (section 3, arrow). NT: neural tube, NC: notochord, CG: cement gland. (TIF 10234 kb

Protocol of a population-based prospective COVID-19 cohort study Munich, Germany (KoCo19)

Radon K, Saathoff E, Pritsch M, et al. Protocol of a population-based prospective COVID-19 cohort study Munich, Germany (KoCo19). BMC Public Health. 2020;20(1): 1036

Socioeconomic and risk-related drivers of compliance with measures to prevent SARS-CoV-2 infection: evidence from the Munich-based KoCo19 study

Pedron S, Laxy M, Radon K, et al. Socioeconomic and risk-related drivers of compliance with measures to prevent SARS-CoV-2 infection: evidence from the Munich-based KoCo19 study. BMC Public Health. 2023;23(1): 860.**Objectives** Although a growing share of the population in many countries has been vaccinated against the SARS-CoV-2 virus to different degrees, social distancing and hygienic non-pharmaceutical interventions still play a substantial role in containing the pandemic. The goal of this study was to investigate which factors are correlated with a higher compliance with these regulations in the context of a cohort study in the city of Munich, southern Germany, during the summer of 2020, i.e. after the first lockdown phase. **Methods** Using self-reported compliance with six regulations and personal hygiene rules (washing hands, avoiding touching face, wearing a mask, keeping distance, avoiding social gatherings, avoiding public spaces) we extracted two compliance factor scores, namelycompliance with personal hygiene measuresandcompliance with social distancing regulations. Using linear and logistic regressions, we estimated the correlation of several socio-demographic and risk perception variables with both compliance scores. **Results** Risk aversion proved to be a consistent and significant driver of compliance across all compliance behaviors. Furthermore, being female, being retired and having a migration background were positively associated with compliance with personal hygiene regulations, whereas older age was related with a higher compliance with social distancing regulations. Generally, socioeconomic characteristics were not related with compliance, except for education, which was negatively related with compliance with personal hygiene measures. **Conclusions** Our results suggest that for a targeted approach to improve compliance with measures to prevent SARS-CoV-2 infection, special attention should be given to younger, male and risk-prone individuals

Regulation of distinct branches of the non-canonical Wnt-signaling network in Xenopus dorsal marginal zone explants

Background: A tight regulation of the Wnt-signaling network, activated by 19 Wnt molecules and numerous receptors and co-receptors, is required for the establishment of a complex organism. Different branches of this Wnt-signaling network, including the canonical Wnt/β-catenin and the non-canonical Wnt/PCP, Wnt/Ror2 and Wnt/Ca$^{2+}$ pathways, are assigned to distinct developmental processes and are triggered by certain ligand/receptor complexes. The Wnt-signaling molecules are closely related and it is still on debate whether the information for activating a specific branch is encoded by specific sequence motifs within a particular Wnt protein. The model organism Xenopus offers tools to distinguish between Wnt-signaling molecules activating distinct branches of the network. Results: We created chimeric Wnt8a/Wnt11 molecules and could demonstrate that the C-terminal part (containing the BS2) of Wnt8a is responsible for secondary axis formation. Chimeric Wnt11/Wnt5a molecules revealed that the N-terminus with the elements PS3-1 and PS3-2 defines Wnt11 specificity, while elements PS3-1, PS3-2 and PS3-3 are required for Wnt5a specificity. Furthermore, we used Xenopus dorsal marginal zone explants to identify non-canonical Wnt target genes regulated by the Wnt5a branch and the Wnt11 branch. We found that pbk was specifically regulated by Wnt5a and rab11fip5 by Wnt11. Overexpression of these target genes phenocopied the overexpression of their regulators, confirming the distinct roles of Wnt11 and Wnt5a triggered signaling pathways. Furthermore, knock-down of pbk was able to restore convergent extension movements in Wnt5a morphants. Conclusions: The N-terminal part of non-canonical Wnt proteins decides whether the Wnt5a or the Wnt11 branch of the Wnt-signaling network gets activated. The different non-canonical Wnt branches not only regulate cellular behavior, but, surprisingly, also regulate the expression of different target genes. One of these target genes, pbk, seems to be the relevant target gene executing Wnt5a-mediated regulation of convergent extension movements