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: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
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: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:We decellularized rat diaphragms with the main detergents 1% or 0.1% sodium dodecyl sulfate (SDS) and 4% sodium deoxycholate (SDC) by orbital shaking (OS) or retrograde perfusion (RP) through the vena cava. Our systematic comparison entails evaluation of decellularized diaphragmatic samples by (1) quantitative analysis including DNA quantification and biomechanical testing, (2) qualitative and semi-quantitative analysis by proteomics, as well as (3) qualitative assessment with macroscopic and microscopic evaluation by histological staining, immunohistochemistry and scanning electron microscopy.
Project description:We created a rat renal congestion model and investigated the effect of renal congestion on hemodynamics and molecular mechanisms. The inferior vena cava (IVC) between the renal veins was ligated by suture in male Sprague-Dawley rats to increase upstream IVC pressure and induce congestion in the left kidney only. Left kidney congestion reduced renal blood flow, glomerular filtration rate, and increased renal interstitial hydrostatic pressure. Tubulointerstitial and glomerular injury and medullary thick ascending limb hypoxia were observed only in the congestive kidneys. Molecules related to extracellular matrix expansion, tubular injury, and focal adhesion were upregulated in microarray analysis. Renal decapsulation ameliorated the tubulointerstitial injury. Electron microscopy captured pericyte detachment in the congestive kidneys. Transgelin and platelet-derived growth factor receptors, as indicators of pericyte-myofibroblast transition, were upregulated in the pericytes and the adjacent interstitium. With the compression of the peritubular capillaries and tubules, hypoxia and physical stress induce pericyte detachment, which could result in extracellular matrix expansion and tubular injury in renal congestion.
Project description:Vena cava and thoracic aorta fragments were dissected out from 12 to 20 Tie2-GFP donor mice and GFP+ endothelial cells were isolatred by FACsorting and collected in RNA lysis buffer for subsequent total RNA isolation. Total RNA was loaded on a chip for microarray.
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.