ABSTRACT: Dlx1 and Rgs5 in the Ductus Arteriosus: Vessel-specific Genes Identified by Transcriptional Profiling of Laser-capture Microdissected Endothelial and Smooth Muscle Cells
Project description:Closure or patency of the ductus arteriosus is a critical event in neonatal life. We aimed to identify genes that are specifically expressed in the ductus arteriosus versus (the non-closing) aorta Gene expression profiling of laser-captured microdissected cells offers the opportunity to study gene expression profiles in cells of different embryonic origin Comparative microarray analysis of laser-capture dissected endothelial and smooth muscle cells of the rat ductus arteriosus and aorta at embryonic age E18 and E21.
Project description:Failure to close the ductus arteriosus immediately post-birth, patent ductus arteriosus (PDA), accounts for up to 10% of all congenital heart defects. Despite significant advances in PDA management options, including pharmacological treatment targeting the prostaglandin pathway, a proportion of patients fail to respond and must undergo surgical intervention. Thus, further refinement of the cellular and molecular mechanisms that govern vascular remodeling of this vessel is required. As anticipated, single-cell RNA sequencing on the ductus arteriosus in mouse embryos at E18.5, P0.5, and P5, revealed broad transcriptional alterations in the endothelial, smooth muscle, and fibroblast cell compartments. After close evaluation of the transcriptomic dataset, we predicted that vimentin might be required for complete closure of the vessel. Subsequent studies demonstrated that, in fact, mice with genetic deletion of vimentin fail to fully remodel the ductus arteriosus. Through single-cell RNA-sequencing and by tracking closure of the ductus arteriosus postnatally in mice, we uncovered the unexpected contribution of vimentin in driving complete closure of the ductus arteriosus potentially through regulation of the Notch signaling pathway.
Project description:Closure or patency of the ductus arteriosus is a critical event in neonatal life. We aimed to identify genes that are specifically expressed in the ductus arteriosus versus (the non-closing) aorta Gene expression profiling of laser-captured microdissected cells offers the opportunity to study gene expression profiles in cells of different embryonic origin
Project description:To investigate the function of PRDM6 in smooth muscle cells during outflow tract development, we have isolated the ascending aorta and the ductus arteriosus (DA) separatly from the E18.5 Wnt1Cre;PRDM6f/f mouse (PRDM6 KO) and PRDM6f/f mouse (Control) and compared the gene expression profile of the four groups of samples
Project description:The ductus arteriosus constricts after birth or hatching and eventually closes to terminate embryonic circulation. Chicken embryos have two long ductus arteriosus. Then the pulmonary artery-sided and descending aorta-sided ductus arteriosus have distinct functional characteristics, such as oxygen responsiveness. We used microarrays to elucidate the difference between the pulmonary artery-sided and descending aorta-sided ductus arteriosus in their transcriptional profiles.
Project description:Patent ductus arteriosus (PDA) is the third most common congenital heart disease and resulted from the persistence of ductal patency after birth. Ductus arteriosus closure involves functional and structural remodeling, controlled by many factors. The closure-related human plasma proteins are unknown. Here we for the first time demonstrate six key differential plasma proteins in the human PDA patients using proteomic technology and present a model to illustrate the constriction and closure of ductus arteriosus. We found that permanent closure might be regulated by the proteins related to platelet activation and coagulation cascades, complement mannan-binding-lectin, and other systemic signaling pathways. Our findings indicate that the differential proteins involved in these pathways may play key roles in the non-closure of the ductus arteriosus and may be developed as biomarkers for diagnosis. All those findings may be served as the basis of understanding the etiology and pathogenesis of PDA.
Project description:This study aims to identify and characterize miRNA expression in aneurysmal smooth muscle cells (SMCs) and M1 and M2 macrophages isolated by laser microdissection from human AAA biopsy samples. The aim of this study was to profile (with microarray technology) miRNAs in M1 and M2 macrophages and Smooth muscle cells (SMCs), isolated by laser capture microdissection (LCM). SMCs isolated from control non-aneurysmal aortas were the control group according to which data were normalized. Areas enriched in M1 macrophages, M2 macrophages and smooth muscle cells were microdissected (9.8 [4.7-16.2] mm2) from two different biopsy samples of human abdominal aortic aneurysm. An area enriched in smooth muscle cells was microdissected from two biopsy samples of non aneurysmal abdominal aortas. Each microdissected samples were analyzed independently on microarray.
Project description:This study aims to identify and characterize miRNA expression inATLOs isolated by laser microdissection from human AAA biopsy samples. The aim of this study was to profile (with microarray technology) miRNAs in ATLOs (Adventitial tertiary lymphoid organs), isolated by laser capture microdissection (LCM). Smooth muscle cells (SMCs) isolated from control non-aneurysmal aortas were the control group according to which data were normalized. ATLOs were microdissected (mean 13.5mm2) from two different biopsy samples of human abdominal aortic aneurysm. An area enriched in smooth muscle cells was microdissected from two biopsy samples of non aneurysmal abdominal aortas. Each microdissected samples were analyzed independently on microarray.
Project description:This study aims to identify and characterize miRNA expression in aneurysmal smooth muscle cells (SMCs) and M1 and M2 macrophages isolated by laser microdissection from human AAA biopsy samples. The aim of this study was to profile (with microarray technology) miRNAs in M1 and M2 macrophages and Smooth muscle cells (SMCs), isolated by laser capture microdissection (LCM). SMCs isolated from control non-aneurysmal aortas were the control group according to which data were normalized. Areas enriched in M1 macrophages, M2 macrophages and smooth muscle cells were microdissected (9.8 [4.7-16.2] mm2) from two different biopsy samples of human abdominal aortic aneurysm. An area enriched in smooth muscle cells was microdissected from two biopsy samples of non aneurysmal abdominal aortas. Each microdissected samples were analyzed independently on microarray. M1 and M2 macrophages and smooth muscle cells expressed in human abdominal aneurysmal aortas were each analyzed in duplicate (each isolated from different human donors of abdominal aortic aneurysm). Analysis of replicated samples of cells were performed on a second microarray. Data were normalized with smooth muscle cells isolated from human abdominal non aneurysmal aortas.
Project description:How two-chambered hearts in basal vertebrates have evolved from single-chamber hearts found in ancestral chordates remains unclear. Here, we show that the teleost sinus venosus (SV) is a chamber-like vessel comprised of an outer layer of smooth muscle cells. We find that in adult zebrafish nr2f1a mutants, which lack atria, the SV comes to physically resemble the thicker bulbus arteriosus (BA) at the arterial pole of the heart through an adaptive, hypertensive response involving smooth muscle proliferation due to aberrant hemodynamic flow. Single cell transcriptomics show that smooth muscle and endothelial cell populations within the adapting SV also take on arterial signatures. Bulk transcriptomics of the blood sinuses flanking the tunicate heart reinforce a model of greater equivalency in ancestral chordate BA and SV precursors. Our data simultaneously reveal that secondary complications from congenital heart defects can develop in adult zebrafish similar to those in humans and that the foundation of equivalency between flanking auxiliary vessels may remain latent within basal vertebrate hearts.
