352 research outputs found
BPA disrupts meiosis I in oogonia by acting on pathways including cell cycle regulation, meiosis initiation and spindle assembly
This work was supported by funding to PAF and CC from the Wellcome Trust (080388) and the European Community's Seventh Framework Programme under grant agreement no. 212885. (http://www.abdn.ac.uk/reef/). The authors would like to thank INRAE, SAAJ, experimental animal facility (Sciences de l'Animal et de l'Aliment de Jouy), especially Jean-Pierre Albert, Didier Mauchand, and Jean-François Alkombre.Peer reviewedPublisher PD
Dynamic CpG methylation delineates subregions within super-enhancers selectively decommissioned at the exit from naive pluripotency.
Clusters of enhancers, referred as to super-enhancers (SEs), control the expression of cell identity genes. The organisation of these clusters, and how they are remodelled upon developmental transitions remain poorly understood. Here, we report the existence of two types of enhancer units within SEs typified by distinctive CpG methylation dynamics in embryonic stem cells (ESCs). We find that these units are either prone for decommissioning or remain constitutively active in epiblast stem cells (EpiSCs), as further established in the peri-implantation epiblast in vivo. Mechanistically, we show a pivotal role for ESRRB in regulating the activity of ESC-specific enhancer units and propose that the developmentally regulated silencing of ESRRB triggers the selective inactivation of these units within SEs. Our study provides insights into the molecular events that follow the loss of ESRRB binding, and offers a mechanism by which the naive pluripotency transcriptional programme can be partially reset upon embryo implantation
Ovine fetal testis stage-specific sensitivity to environmental chemical mixtures
Acknowledgements We thank George Corsar and Jim MacDonald for the management of experimental animals Funding This work was supported by the European Commission Framework 7 Programme (Contract No 212885)Peer reviewedPublisher PD
Evolution of T cell receptor beta loci in salmonids
T-cell mediated immunity relies on a vast array of antigen specific T cell receptors (TR). Characterizing the structure of TR loci is essential to study the diversity and composition of T cell responses in vertebrate species. The lack of good-quality genome assemblies, and the difficulty to perform a reliably mapping of multiple highly similar TR sequences, have hindered the study of these loci in non-model organisms. High-quality genome assemblies are now available for the two main genera of Salmonids, Salmo and Oncorhynchus. We present here a full description and annotation of the TRB loci located on chromosomes 19 and 25 of rainbow trout (Oncorhynchus mykiss). To get insight about variations of the structure and composition of TRB locus across salmonids, we compared rainbow trout TRB loci with other salmonid species and confirmed that the basic structure of salmonid TRB locus is a double set of two TRBV-D-J-C loci in opposite orientation on two different chromosomes. Our data shed light on the evolution of TRB loci in Salmonids after their whole genome duplication (WGD). We established a coherent nomenclature of salmonid TRB loci based on comprehensive annotation. Our work provides a fundamental basis for monitoring salmonid T cell responses by TRB repertoire sequencing
Viral Resistance and IFN Signaling in STAT2 Knockout Fish Cells
Copyright © 2019 The Authors.Peer reviewedPublisher PD
Porcine Reproductive and Respiratory Syndrome Virus Type 1.3 Lena Triggers Conventional Dendritic Cells 1 Activation and T Helper 1 Immune Response Without Infecting Dendritic Cells
Porcine Reproductive and Respiratory Syndrome virus (PRRSV) is an arterivirus responsible for highly contagious infection and huge economic losses in pig industry. Two species, PRRSV-1 and PRRSV-2 are distinguished, PRRSV-1 being more prevalent in Europe. PRRSV-1 can further be divided in subtypes. PRRSV-1.3 such as Lena are more pathogenic than PRRSV-1.1 such as Lelystad or Flanders13. PRRSV-1.3 viruses trigger a higher Th1 response than PRRSV-1.1, although the role of the cellular immune response in PRRSV clearance remains ill defined. The pathogenicity as well as the T cell response inductions may be differentially impacted according to the capacity of the virus strain to infect and/or activate DCs. However, the interactions of PRRSV with in vivo-differentiated-DC subtypes such as conventional DC1 (cDC1), cDC2, and monocyte-derived DCs (moDC) have not been thoroughly investigated. Here, DC subpopulations from Lena in vivo infected pigs were analyzed for viral genome detection. This experiment demonstrates that cDC1, cDC2, and moDC are not infected in vivo by Lena. Analysis of DC cytokines production revealed that cDC1 are clearly activated in vivo by Lena. In vitro comparison of 3 Europeans strains revealed no infection of the cDC1 and cDC2 and no or little infection of moDC with Lena, whereas the two PRRSV-1.1 strains infect none of the 3 DC subtypes. In vitro investigation of T helper polarization and cytokines production demonstrate that Lena induces a higher Th1 polarization and IFNγ secretion than FL13 and LV. Altogether, this work suggests an activation of cDC1 by Lena associated with a Th1 immune response polarization
Separated children seeking asylum in Ireland.
This report updates the first report of the Irish Refugee Council published in 1999, entitled Separated children seeking asylum in Ireland: A report on legal and social conditions. At the time of the publication of that report, there were 32 separated children seeking asylum in Ireland. The number of separated children seeking asylum in Ireland has increased markedly. By March 2003, the number of separated children, entering Ireland and referred to the North Eastern Area Health Board was 2,7172. Nearly half, or 1,113 children, were reunited with family members already in Ireland. 1,316 separated children, under the care of the Health Boards, have made applications for asylum under the 1951 Geneva Convention on the Status of Refugees. Neither the Government nor non-statutory agencies anticipated this increase in the numbers of separated minors arriving in Ireland. Therefore administrative procedures and care services have had to be responsive to emergent needs rather than having developed through advance planning. This report aims to examine policy and practice with respect to the legal and social conditions of separated children in Ireland, in light of the Separated Children in Europe Programme’s (SCEP)3 ‘Statement of Good Practice’ (SGP). The Irish Refugee Council, a member of the Separated Children in Europe Programme, commissioned the report
Prolonged dialysis during ex vivo lung perfusion promotes inflammatory responses
Ex-vivo lung perfusion (EVLP) has extended the number of transplantable lungs by reconditioning marginal organs. However, EVLP is performed at 37°C without homeostatic regulation leading to metabolic wastes’ accumulation in the perfusate and, as a corrective measure, the costly perfusate is repeatedly replaced during the standard of care procedure. As an interesting alternative, a hemodialyzer could be placed on the EVLP circuit, which was previously shown to rebalance the perfusate composition and to maintain lung function and viability without appearing to impact the global gene expression in the lung. Here, we assessed the biological effects of a hemodialyzer during EVLP by performing biochemical and refined functional genomic analyses over a 12h procedure in a pig model. We found that dialysis stabilized electrolytic and metabolic parameters of the perfusate but enhanced the gene expression and protein accumulation of several inflammatory cytokines and promoted a genomic profile predicting higher endothelial activation already at 6h and higher immune cytokine signaling at 12h. Therefore, epuration of EVLP with a dialyzer, while correcting features of the perfusate composition and maintaining the respiratory function, promotes inflammatory responses in the tissue. This finding suggests that modifying the metabolite composition of the perfusate by dialysis during EVLP can have detrimental effects on the tissue response and that this strategy should not be transferred as such to the clinic
Transcriptional Responses of Resistant and Susceptible Fish Clones to the Bacterial Pathogen Flavobacterium psychrophilum
Flavobacterium psychrophilum is a bacterial species that represents one of the most important pathogens for aquaculture worldwide, especially for salmonids. To gain insights into the genetic basis of the natural resistance to F. psychrophilum, we selected homozygous clones of rainbow trout with contrasted susceptibility to the infection. We compared the transcriptional response to the bacteria in the pronephros of a susceptible and a resistant line by micro-array analysis five days after infection. While the basal transcriptome of healthy fish was significantly different in the resistant and susceptible lines, the transcriptome modifications induced by the bacteria involved essentially the same genes and pathways. The response to F. psychrophilum involved antimicrobial peptides, complement, and a number of enzymes and chemokines. The matrix metalloproteases mmp9 and mmp13 were among the most highly induced genes in both genetic backgrounds. Key genes of both pro- and anti-inflammatory response such as IL1 and IL10, were up-regulated with a greater magnitude in susceptible animals where the bacterial load was also much higher. While higher resistance to F. psychrophilum does not seem to be based on extensive differences in the orientation of the immune response, several genes including complement C3 showed stronger induction in the resistant fish. They may be important for the variation of susceptibility to the infection
A multi-scale analysis of bull sperm methylome revealed both species peculiarities and conserved tissue-specific
peer-reviewedBackground: Spermatozoa have a remarkable epigenome in line with their degree of specialization, their unique
nature and different requirements for successful fertilization. Accordingly, perturbations in the establishment of DNA
methylation patterns during male germ cell differentiation have been associated with infertility in several species.Background: Spermatozoa have a remarkable epigenResults: The quantification of DNA methylation at CCGG sites using luminometric methylation assay (LUMA)
highlighted the undermethylation of bull sperm compared to the sperm of rams, stallions, mice, goats and men.
Total blood cells displayed a similarly high level of methylation in bulls and rams, suggesting that undermethylation
of the bovine genome was specific to sperm. Annotation of CCGG sites in different species revealed no striking bias
in the distribution of genome features targeted by LUMA that could explain undermethylation of bull sperm. To
map DNA methylation at a genome-wide scale, bull sperm was compared with bovine liver, fibroblasts and
monocytes using reduced representation bisulfite sequencing (RRBS) and immunoprecipitation of methylated DNA
followed by microarray hybridization (MeDIP-chip). These two methods exhibited differences in terms of genome
coverage, and consistently, two independent sets of sequences differentially methylated in sperm and somatic cells
were identified for RRBS and MeDIP-chip. Remarkably, in the two sets most of the differentially methylated
sequences were hypomethylated in sperm. In agreement with previous studies in other species, the sequences that
were specifically hypomethylated in bull sperm targeted processes relevant to the germline differentiation program
(piRNA metabolism, meiosis, spermatogenesis) and sperm functions (cell adhesion, fertilization), as well as satellites
and rDNA repeats.
Conclusions: These results highlight the undermethylation of bull spermatozoa when compared with both bovine
somatic cells and the sperm of other mammals, and raise questions regarding the dynamics of DNA methylation in
bovine male germline. Whether sperm undermethylation has potential interactions with structural variation in the
cattle genome may deserve further attention.
While bull semen is widely used in artificial insemination, the literature describing DNA methylation in bull
spermatozoa is still scarce. The purpose of this study was therefore to characterize the bull sperm methylome
relative to both bovine somatic cells and the sperm of other mammals through a multiscale analysis
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