The bacterial artificial chromosome (BAC) system is widely used in isolation of large genomic fragments of interest. Construction of a routine BAC library requires several months for picking clones and arraying BACs into superpools in order to employ 4D-PCR to screen positive BACs, which might be time-consuming and laborious. The major histocompatibility complex (MHC) is a cluster of genes involved in the vertebrate immune system, and the classical avian MHC-B locus is a minimal essential one, occupying a 100-kb genomic region. In this study, we constructed a more effective reverse-4D BAC library for the golden pheasant, which first creates sub-libraries and then only picks clones of positive sub-libraries, and identified several MHC clones within thirty days. The full sequencing of a 97-kb reverse-4D BAC demonstrated that the golden pheasant MHC-B locus contained 20 genes and showed good synteny with that of the chicken. The notable differences between these two species were the numbers of class II B loci and NK genes and the inversions of the TAPBP gene and the TAP1-TAP2 region. Furthermore, the inverse TAP2-TAP1 was unique in the golden pheasant in comparison with that of chicken, turkey, and quail. The newly defined genomic structure of the golden pheasant MHC will give an insight into the evolutionary history of the avian MHC.
Our knowledge of the Major Histocompatibility Complex (MHC) in birds is limited because it often consists of numerous duplicated genes within individuals that are difficult to assemble with short read sequencing technology. Long-read sequencing provides an opportunity to overcome this limitation because it allows the assembly of long regions with repetitive elements. In this study we used genomes based on long-read sequencing to predict the number and location of MHC loci in a broad range of bird taxa. From the long-read based genomes of 34 species, we found that there was extremely large variation in the number of MHC loci between species. Overall, there were greater numbers of both class I and II loci in passerines than non-passerines. The highest numbers of loci (up to 193 class II loci) were found in manakins (Pipridae), which had previously not been studied at the MHC. Our results provide the first direct evidence from passerine genomes of this high level of duplication. We also found different duplication patterns between species. In some species both MHC class I and II genes were duplicated together, while in most species they were duplicated independently. Our study shows that the analysis of long-read based genomes can dramatically improve in our knowledge of MHC structure, although further improvements in chromosome level assembly are needed to understand the evolutionary mechanisms producing the extraordinary inter-specific variation in the architecture of the MHC region.
Background: STK40 is a putative serine/threonine kinase. Little is known about its physiological function. Results: STK40 deletion causes fetal lung immaturity and neonatal death with altered expression of genes important for lung development. Conclusion: STK40 is required for alveolar epithelium maturation and function. Significance: Discovery of the role of STK40 in lung maturation helps to understand the molecular regulation in lung development.
Broodiness is observed in most domestic fowls and influences egg production. The goose is one of the most important waterfowls, having strong broody behavior. However, whether autophagy and follicular internal environment play a role in the broodiness behavior of goose is unknown. In this report, we analyzed the follicular internal environment and granulosa cell autophagy of goose follicles. The results show that the contents of hormones, including prolactin (PRL), progesterone (P4), and estradiol (E2), increased in broody goose follicles. Most importantly, the level of granulosa cell autophagy in broody goose follicles was elevated, detected by electron microscopy and western blotting. Also, the expressions of positive regulators of autophagy, including miR-7, miR-29, miR-100, miR-181, PRLR, LC3, p53,Beclin1, Atg9, and Atg12, were up-regulated and the expressions of negative regulators of autophagy, including miR-34b and miR-34c, were down-regulated in broody goose follicles. Our results suggest that goose broodiness is involved in increased granulosa cell autophagy and homeostasis imbalance of internal environment in the follicles. This work contributes to our knowledge of goose broodiness and may influence egg production.
Habitat fragmentation can reduce population viability, especially for area-sensitive species. The Minimum Area Requirements (MAR) of a population is the area required for the population’s long-term persistence. In this study, the response of occupancy probability of giant pandas against habitat patch size was studied in five of the six mountain ranges inhabited by giant panda, which cover over 78% of the global distribution of giant panda habitat. The probability of giant panda occurrence was positively associated with habitat patch area, and the observed increase in occupancy probability with patch size was higher than that due to passive sampling alone. These results suggest that the giant panda is an area-sensitive species. The MAR for giant panda was estimated to be 114.7 km2 based on analysis of its occupancy probability. Giant panda habitats appear more fragmented in the three southern mountain ranges, while they are large and more continuous in the other two. Establishing corridors among habitat patches can mitigate habitat fragmentation, but expanding habitat patch sizes is necessary in mountain ranges where fragmentation is most intensive.
It is important to simulate the entire-process collapse of earthquake excited cable-stayed bridge in order to assure the damage extent under strong earthquakes and to optimize the anti-collapse seismic measures. The newly developed vector form intrinsic finite element (VFIFE) method is capable of computing large deformation, large displacement, collision, and fractures that would happen in the structural collapse. Therefore, it has the potential to simulate the entire-process collapse of earthquake excited structures. For the cable-stayed bridges, the axial forces change significantly during the earthquake excitation. The interaction between the axial forces and bending moments should be taken into account. In order to address this issue, this paper first presents the formulation for integration of VFIFE method and fiber beam-column element model. A bilinear elastic-plastic constitutive damage model is proposed for the mechanical behavior of the fiber and element. The collapse analysis of a 2D mockup bridge scaled from an actual cable-stayed bridge is then conducted. The entire process of the structural damage and collapse are successfully simulated by the proposed method VFIFE-Fiber. This study provides a foundation for seismic damage prediction and anti-collapse seismic design for cable-stayed bridges.
A better understanding of molecular regulation in adipogenesis might help the development of efficient strategies to cope with obesity-related diseases. Here, we report that CCAAT/enhancer-binding protein (C/EBP) β and C/EBPδ, two crucial pro-adipogenic transcription factors, are controlled at a translational level by serine/threonine kinase 40 (Stk40). Genetic knockout (KO) or knockdown (KD) of Stk40 leads to increased protein levels of C/EBP proteins and adipocyte differentiation in mouse embryonic fibroblasts (MEFs), fetal liver stromal cells, and mesenchymal stem cells (MSCs). In contrast, overexpression of Stk40 abolishes the enhanced C/EBP protein translation and adipogenesis observed in Stk40-KO and -KD cells. Functionally, knockdown of C/EBPβ eliminates the enhanced adipogenic differentiation in Stk40-KO and -KD cells substantially. Mechanistically, deletion of Stk40 enhances phosphorylation of eIF4E-binding protein 1, leading to increased eIF4E-dependent translation of C/EBPβ and C/EBPδ. Knockdown of eIF4E in MSCs decreases translation of C/EBP proteins. Moreover, Stk40-KO fetal livers display an increased adipogenic program and aberrant lipid and steroid metabolism. Collectively, our study uncovers a new repressor of C/EBP protein translation as well as adipogenesis and provides new insights into the molecular mechanism underpinning the adipogenic program.
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