RESUMEN
How left-right (LR) asymmetry emerges in a patterning field along the anterior-posterior axis remains an unresolved problem in developmental biology. Left-biased Nodal emanating from the LR organizer propagates from posterior to anterior (PA) and establishes the LR pattern of the whole embryo. However, little is known about the regulatory mechanism of the PA spread of Nodal and its asymmetric activation in the forebrain. Here, we identify bilaterally expressed Follistatin (Fst) as a regulator blocking the propagation of the zebrafish Nodal ortholog Southpaw (Spaw) in the right lateral plate mesoderm (LPM), and restricting Spaw transmission in the left LPM to facilitate the establishment of a robust LR asymmetric Nodal patterning. In addition, Fst inhibits the Activin-Nodal signaling pathway in the forebrain thus preventing Nodal activation prior to the arrival, at a later time, of Spaw emanating from the left LPM. This contributes to the orderly propagation of asymmetric Nodal activation along the PA axis. The LR regulation function of Fst is further confirmed in chick and frog embryos. Overall, our results suggest that a robust LR patterning emerges by counteracting a Fst barrier formed along the PA axis.
Asunto(s)
Proteínas de Pez Cebra , Pez Cebra , Animales , Pez Cebra/genética , Pez Cebra/metabolismo , Proteínas de Pez Cebra/genética , Proteínas de Pez Cebra/metabolismo , Folistatina/genética , Folistatina/metabolismo , Tipificación del Cuerpo/genética , Factor de Crecimiento Transformador beta/metabolismo , Regulación del Desarrollo de la Expresión GénicaRESUMEN
Feingold syndrome type 1, caused by loss-of-function of MYCN, is characterized by varied phenotypes including esophageal and duodenal atresia. However, no adequate model exists for studying the syndrome's pathological or molecular mechanisms, nor is there a treatment strategy. Here, we developed a zebrafish Feingold syndrome type 1 model with nonfunctional mycn, which had severe intestinal atresia. Single-cell RNA-seq identified a subcluster of intestinal cells that were highly sensitive to Mycn, and impaired cell proliferation decreased the overall number of intestinal cells in the mycn mutant fish. Bulk RNA-seq and metabolomic analysis showed that expression of ribosomal genes was down-regulated and that amino acid metabolism was abnormal. Northern blot and ribosomal profiling analysis showed abnormal rRNA processing and decreases in free 40S, 60S, and 80S ribosome particles, which led to impaired translation in the mutant. Besides, both Ribo-seq and western blot analysis showed that mTOR pathway was impaired in mycn mutant, and blocking mTOR pathway by rapamycin treatment can mimic the intestinal defect, and both L-leucine and Rheb, which can elevate translation via activating TOR pathway, could rescue the intestinal phenotype of mycn mutant. In summary, by this zebrafish Feingold syndrome type 1 model, we found that disturbance of ribosomal biogenesis and blockage of protein synthesis during development are primary causes of the intestinal defect in Feingold syndrome type 1. Importantly, our work suggests that leucine supplementation may be a feasible and easy treatment option for this disease.
Asunto(s)
Microcefalia , Pez Cebra , Animales , Proteína Proto-Oncogénica N-Myc , Pez Cebra/metabolismo , Microcefalia/genética , Serina-Treonina Quinasas TOR/metabolismo , LeucinaRESUMEN
Powdery mildew (PM) significantly and negatively affects the yield and quality of melon (Cucumis melo) worldwide. Race 2F is the predominant physiological race of the pathogen Podosphaera xanthii in many regions. We used accessions PMR 6 (P1; resistant to PM) and M1-7 (P2; susceptible to PM) to analyze the inheritance of resistance to PM (race 2F). The ratio between resistant and susceptible individuals fits a Mendelian segregation ratio of 13:3 in a total of 256 F2 individuals and 1:1 in BC1P2. The resistance to PM in PMR 6 was governed by two genes: a dominant (AA) gene with an epistatic effect and a recessive gene (bb). Only individuals with aaBB or aaBb genotypes were susceptible to PM. Two PM resistance loci, Pm2.1 and pm12.1, were mapped on chromosomes 2 and 12 by bulked segregant analysis and secondary mapping by quantitative trait loci analysis with 18 markers. A new marker-assisted selection system to identify melon genotypes resistant or susceptible to PM was developed and tested in 93 melon accessions. Nucleotide diversity (π) and fixation index (Fst) for the two PM resistance loci were estimated using resequencing data of 336 melons from three groups: C. melo subsp. agrestis, Cucumis melo subsp. melo, and the intermediate type. The lowest π was observed in C. melo ssp. agrestis, and the highest Fst value was between C. melo ssp. agrestis and C. melo ssp. melo. The findings provide a promising tool that can be used to accelerate breeding for durable resistance to PM.
Asunto(s)
Cucumis melo , Cucurbitaceae , Ascomicetos , Cucumis melo/genética , Nucleótidos , Enfermedades de las PlantasRESUMEN
PURPOSE: Electroencephalography (EEG) remains the gold standard for identifying rhythmic and periodic patterns in critically ill patients. Residents have frequent exposures to EEG and critically ill patients during their training. Our study aimed to assess resident competency in the use of the American Clinical Neurophysiology Society (ACNS) critical care EEG terminology. METHODS: After self-guided reading and a 2-hour session reviewing the ACNS critical care EEG Terminology training slides, 16 adult neurology residents (PGY 2-4) completed the ACNS certification test. Performance scores were reported as average percent agreement (PA%) with a previously established 5-member expert panel. Interrater agreement was calculated to gauge consensus among peers within the resident cohort. Self-reported comfort levels using the terminology were also obtained. RESULTS: The overall pass rate for our cohort was 50% and the median score was 74%. The terms with the highest PA% were: seizures (86.4%), main term 1 (78%), main term 2 (74%). Interrater agreement scores (kappa values) were almost perfect for seizure, and substantial for main terms 1 and 2. CONCLUSIONS: Our data suggests that with minimal investment, adult neurology residents at various stages of training can effectively learn the ACNS critical care EEG Terminology.