Arterial vs venous expression of genes: differences in inflammatory genes
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ABSTRACT: Arteries and veins modulate cardiovascular homeostasis and contribute to the pathogenesis of hypertension. Functional differences between normal arteries and veins are based upon differences in gene expression. To better characterize these expression patterns, and to identify candidate genes that could be manipulated selectively in the venous system, we performed whole genome expression profiling of rat arteries and veins using the CodeLink platform. We used the major artery and vein of the rat, the thoracic aorta and caudal vena cava, respectively. Expression of mRNA for thrombospondins (TSP-1, 2, 4) was greater than 5-fold higher in veins vs arteries. The most prominent gene expression difference between the normal aorta and vena cava was pancreatitis associated protein (PAP1), a protein with anti-inflammatory functions that was 64-fold higher in vena cava vs aorta. Higher mRNA expression of TSP-1, TSP-2, TSP-4 and PAP1 in vena cava vs aorta was confirmed with real time RT-PCR. Importantly, immunohistochemical analysis of blood vessels sections qualitatively confirmed a higher expression of proteins in vena cava vs aorta. These studies report a difference in inflammatory genes in arteries vs veins. A particularly notable finding is the discovery of PAP1 mRNA and protein expression in peripheral blood vessels with a substantially higher expression in the veins. Data from these studies may provide novel insights into the genetic basis for functional differences between arteries and veins in health and disease.
Project description:Arteries and veins modulate cardiovascular homeostasis and contribute to the pathogenesis of hypertension. Functional differences between normal arteries and veins are based upon differences in gene expression. To better characterize these expression patterns, and to identify candidate genes that could be manipulated selectively in the venous system, we performed whole genome expression profiling of rat arteries and veins using the CodeLink platform. We used the major artery and vein of the rat, the thoracic aorta and caudal vena cava, respectively. Expression of mRNA for thrombospondins (TSP-1, 2, 4) was greater than 5-fold higher in veins vs arteries. The most prominent gene expression difference between the normal aorta and vena cava was pancreatitis associated protein (PAP1), a protein with anti-inflammatory functions that was 64-fold higher in vena cava vs aorta. Higher mRNA expression of TSP-1, TSP-2, TSP-4 and PAP1 in vena cava vs aorta was confirmed with real time RT-PCR. Importantly, immunohistochemical analysis of blood vessels sections qualitatively confirmed a higher expression of proteins in vena cava vs aorta. These studies report a difference in inflammatory genes in arteries vs veins. A particularly notable finding is the discovery of PAP1 mRNA and protein expression in peripheral blood vessels with a substantially higher expression in the veins. Data from these studies may provide novel insights into the genetic basis for functional differences between arteries and veins in health and disease. Whole genome expression profiling of aorta and vena cava whole tissues from normal male Sprague-Dawley rats (6 biological replicates each) was performed using the CodeLink Rat Whole Genome Bioarrays.
Project description:Tissue-engineered veins were generated by reconditioning decellularized veins from both human and pig with whole-blood from respective species. Decellularized human vena femoralis from three donor were reconditioned with human whole blood from four donors. In addition, decellularized pig vena cava from three donors were reconditioned with pig whole blood from three different donors. The proteomes of the tissue-engineered veins were investigated applying the TMT-based proteomics to explore differences between species, regarding the gain of biological material by the reconditioning process.
Project description:A porcine microarray study of right ventricular failure due to coronary artery ligation of the right ventricular free wall and subsequent treatment of right ventricular failure by volume unloading using a shunt between superior vena cava and the pulmonary artery (Glenn-shunt) 1. Surgical preparation with a 12 mm graft between superior vena cava and pulmonary artery, the graft is then clamped - Baseline sample using a biopsy needle. 2. After surgical preparation the coronary arteries of the right ventricular free wall are ligated, then heart failure develops over 120 minutes - Failure sample using a biopsy needle. 3. The shunt is then opened and the superior vena cava closed between the shunt and right atrium, diverting the blood from superior vena cava through the shunt for a period of 15 minutes partially unloading the right ventricle - Shunt sample using a biopsy needle. A series of six pigs, three samples from each animal: baseline, failure and shunt/treatment.
Project description:90K Combimatrix gene expression microarrays were composed by 17,048 replicated probes and 963 not replicated specific for the Ensembl (Ver. 56) Transcripts (protein coding + pseudogenes + retrotransposed elements), 11,363 replicated probes specific for the UniGene clusters (Ver. 38) of length comprised between 778 nt and 1,348 nt, and 28,790 single probes specific for the remaining UniGene clusters. These microarrays were used to study gene expression in 9 different pig tissues involved in the cardiocirculatory system (Left Atrium; Right Ventricle, Inferior Vena Cava, Superior Vena Cava, Ascending Aorta, Descending Aorta, Pulmonary Artery, Coronary Artery, Leaflet of Pulmonary Artery). Gene expression was correlated with the micro-RNA expression in the same tissues, also deposited at ArrayExpress under accession number E-MTAB-1938 (https://www.ebi.ac.uk/arrayexpress/experiments/E-MTAB-1938).
Project description:Canine primary endothelial cells (ECs) and vascular smooth muscle cells (vSMCs) were isolated from the vena cava, vena porta and aorta. All tissue sources were derived from three donors for accurate comparison and to reduce inter-animal variation. We used a recently established technique for the isolation of primary ECs with a lesser chance of contamination that can also be used on small vessels. This same vessel was also used for the isolation of vSMCs
Project description:To investigate the effect of volume overload (VO) during postnatal right atrium (RA) development, we established the RA VO model by conducting the fistula between abdominal aorta and inferior vena cava (ACF) on postnatal day 7(P7) C57/BL6 mice. We then performed gene expression analysis using data obtained from RNA-seq of RA from VO and sham-operated mice at postnatal day 21 (P21).
Project description:While blood vessels have muscular walls that undergo tonic contractions to alter vascular resistance and, thus, control blood flow, lymphatics at the level of the collecting vessels and higher have muscular walls capable of rapid phasic contractions that generate lymph flow in addition to tonic contractions that regulate lymph flow resistance. While the ability of lymphatics to undergo rapid phasic contractions has been known for several centuries, the biological elements governing this phenomenon remain unknown. In an attempt to gain insight into the structural and regulatory elements that give lymphatic vessels their unique contractile capabilities, we utilized two-color microarray analysis to compare the thoracic duct of the rat to the vena cava of the same donor animal. Total cellular RNA was isolated immediately following vessel isolation and amplified in the presence of amino allyl dUTP. The resulting modified aRNA was conjugated to either Cy3 or Cy5 dye prior to hybridization to a rat 5.7K oligonucleotide array. Analysis and filtering of the data obtained from the microarray image yielded several contractile and regulatory genes with altered expression in the thoracic duct relative to the vena cava. Further evaluation of the data obtained in this study may aid in illustrating the unique properties of the lymphatic vessel and its muscular wall. Keywords: Thoracic duct, lymphatics, microarray Four unique thoracic duct/vena cava sample pairs were individually analyzed via two-color microarray analysis yielding 4 biological replicates. To minimize dye bias, a dye balance design was utilized in which the orientation of dye assignment was alternated between vessel pairs (i.e. two thoracic duct samples were labeled with Cy3 and two were labeled with Cy5). Prior to analysis, the data from 2 of the replicates was transformed to accomodate the dye balance such that all thoracic duct data is interpreted as Cy5 and all vena cava data is interpreted as Cy3.
Project description:We applied a new computational approach to predict specie-specific and conserved miRNAs, than experimentally confirmed by a modified RNA-primed Array-based Klenow Extension (RAKE) method. We identified 489 conserved and 1,178 pig-specific novel miRNAs increasing our tally of confirmed miRNAs to 1,667 novel miRNAs. In addition, RAKE allowed the identification of miRNA isoforms (isomiRs) that we demonstrated to be differentially expressed across tissues suggesting that subtle variability in isomiR expression is regulated and biologically meaningful. 12K microarray was hybridized with a pool of small RNAs from 20 different tissues (superior vena cava, adipose tissue, lung, spleen, stomach, liver, intestine, kidney, descending aorta, left atrium, left ventricle, skeletal muscle, pulmonary aorta, skin, tongue, ascending aorta, arterial white cells blood, venal white cells blood, coronary valve, lymph node). Same experiment was replicated three times to grant the reliability of the identified 5M-bM-^@M-^Y end of miRNAs and microarray slides were scanned three times at low, medium and high PMT to improve microarray dynamic range.
Project description:We applied a new computational approach to predict specie-specific and conserved miRNAs, than experimentally confirmed by a modified RNA-primed Array-based Klenow Extension (RAKE) method. We identified 489 conserved and 1,178 pig-specific novel miRNAs increasing our tally of confirmed miRNAs to 1,667 novel miRNAs. In addition, RAKE allowed the identification of miRNA isoforms (isomiRs) that we demonstrated to be differentially expressed across tissues suggesting that subtle variability in isomiR expression is regulated and biologically meaningful. 90K microarray was hybridized with a pool of small RNAs from 20 different tissues (superior vena cava, adipose tissue, lung, spleen, stomach, liver, intestine, kidney, descending aorta, left atrium, left ventricle, skeletal muscle, pulmonary aorta, skin, tongue, ascending aorta, arterial white cells blood, venal white cells blood, coronary valve, lymph node). Same experiment was replicated four times to grant the reliability of the identified 3M-bM-^@M-^Y end of miRNAs and microarray slides were scanned three times at low, medium and high PMT to improve microarray dynamic range.
Project description:We applied a new computational approach to predict specie-specific and conserved miRNAs, than experimentally confirmed by a modified RNA-primed Array-based Klenow Extension (RAKE) method. We identified 489 conserved and 1,178 pig-specific novel miRNAs increasing our tally of confirmed miRNAs to 1,667 novel miRNAs. In addition, RAKE allowed the identification of miRNA isoforms (isomiRs) that we demonstrated to be differentially expressed across tissues suggesting that subtle variability in isomiR expression is regulated and biologically meaningful. 90K microarray was hybridized with a pool of small RNAs from 20 different tissues (superior vena cava, adipose tissue, lung, spleen, stomach, liver, intestine, kidney, descending aorta, left atrium, left ventricle, skeletal muscle, pulmonary aorta, skin, tongue, ascending aorta, arterial white cells blood, venal white cells blood, coronary valve, lymph node). Same experiment was replicated four times to grant the reliability of the identified 3M-bM-^@M-^Y end of miRNAs and microarray slides were scanned three times at low, medium and high PMT to improve microarray dynamic range.