Name: Standardized Rat Coronary Ring Preparation and Real-Time Recording of Dynamic Tension Changes Along Vessel Diameter (Video) | JoVE Journal
Uploaded: 2022-06-16T14:50:00.000+00:00
Duration: 7 min 53 s
Description: Learn the wire myograph technique for measuring vascular reactivity in rat coronary arteries. This standardized method enables stable recording of dynamic tension changes in vessels from 60 micrometers to 10 millimeters.

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5 minutes

This work was supported by the Key R&#38;D project of Sichuan Provincial Science and Technology Plan (2022YFS0438), the National Natural Science Foundation of China (82104533), the China Postdoctoral Science Foundation (2020M683273), and the Science &#38; Technology Department of Sichuan Province (2021YJ0175).

The animal protocol was reviewed and approved by the Management Committee from Chengdu University of Traditional Chinese Medicine (Record No. 2021-11). Male Sprague Dawley (SD) rats (260-300 g, 8-10 weeks old) were used for the present study. The rats were kept in an animal chamber and were free to drink and eat during the experiment.
1. Solution preparation
<ol>
	<li>Prepare physiological salt solution (PSS) by dissolving 118 mM of NaCl, 4.7 mM of K+, 2.5 mM of CaCl2, 1.2 mM of KH2PO4, 1.2 mM of MgCl2&#8729;6H2O, 25 mM of NaHCO3, 11 mM of D-glucose, and 5 mM of HEPES (see Table of Materials).</li>
	<li>Prepare high K+ salt solution by dissolving 58 mM of NaCl, 60 mM of K+, 2.5 mM of CaCl2, 1.2 mM of KH2PO4, 1.2 mM of MgCl2&#8729;6H2O, 25 mM of NaHCO3, 11 mM of D-glucose, and 5 mM of HEPES.</li>
	<li>Saturate the above two solutions and bubble with a mixed gas of 95% O2 and 5% CO2. Meanwhile, maintain the pH values of the solution between 7.38 and 7.42 with 2 mM NaOH. 
	NOTE: For details information on solution preparation, please see reference15.</li>
</ol>
2. Rat coronary artery dissection
<ol>
	<li>Anesthetize the rat by inhalation of 2% isoflurane. Confirm deep anesthesia by toe pinch and, if needed, administer additional anesthetics. Then immediately open the thoracic cavity to expose the heart on the portable operating table following a previously published report12.</li>
	<li>After dissociating and removing the heart, drain the residual blood from all the heart chambers by mildly squeezing with medical plastic forceps. Quickly place the pre-processed heart in a Petri dish containing 95% O2 + 5% CO2 saturated PSS at 4 &#176;C, having a pH value of 7.40.</li>
	<li>To accurately identify the anatomical position of the coronary arteries, adjust the posture of the isolated heart under the light microscope according to the schematic diagram (Figure 2A). 
	NOTE: On the frontal view, the right auricle and the pulmonary artery were on the upper left and the upper right, respectively.
	<ol>
		<li>Cut the left and right ventricular cavities along the interventricular septum from the root of the pulmonary artery with surgical scissors and tweezers (Figure 2B).</li>
	</ol>
	</li>
	<li>To dissociate the left and right coronary arteries from the myocardial tissue, dissect the right ventricle under an optical anatomic microscope to thoroughly expose the right coronary artery branch. Then identify the position of the left coronary artery by rotating the heart tissue 45&#176; clockwise (Figure 2D).</li>
	<li>After removing the surrounding sticky myocardial tissue, explicitly discern the pulsing left (about 5 mm) and right (about 5 mm) coronary arteries. Separate the coronary arteries in the middle immediately and completely immerse in PSS at 4 &#176;C. Acquire an arterial ring of about 2 mm by vertically cutting the detached artery with anatomical scissors to record the vascular tension under different stimuli (Figure 2E).</li>
</ol>
3. Suspension and fixation of arterial ring
NOTE: For details on this step, please see reference14.
<ol>
	<li>Prepare two 2 cm stainless steel wires (see Table of Materials) and pre-soak in 4 &#176;C PSS solution saturated with 95% O2 + 5% CO2. Pass both the wires parallelly through the arterial ring along with the vessel&#39;s direction under an optical anatomical microscope and with wires of equal length exposed at both ends of the vascular cavity.</li>
	<li>Fix the arterial ring with the steel wire front and back in the bath of the wire micrograph filled with bubbling PSS with 95% O2 + 5% CO2. Rotate the horizontal screw knob for an appropriate front and rear spacing so that the two wires are horizontal and the arterial ring is in a natural state of relaxation.</li>
	<li>After installing the DMT bath on the thermostatic apparatus, open the data acquisition software (see Table of Materials) to ensure that the corresponding path signal was recorded. Set the following parameters: eyepiece calibration (mm/div): 0.36; target pressure (kPa): 13.3; IC1/IC100: 0.9; online averaging time: 2 s; delay time: 60 s. The steps of arterial ring fixation are shown in Figure 3.</li>
</ol>
4. Standardization of vascular tension in rat arterial ring
NOTE: For different cavity samples, optimal initial tension was necessary for vessels to maintain exceptional activity in vitro. For details, please see reference15.
<ol>
	<li>Achieve the optimal initial tension of the arterial ring by applying a reasonable tension along the diameter of the vessel. 
	NOTE: Based on the previous study16, the maximal agonist-induced tension was accomplished at the factor k value of 0.90 with the initial stretch tension of 1.16 &#177; 0.04 mN/mm (reference values for different vessel samples: k value, 0.90-0.95; initial tension, 1.16-1.52 mN/mm).</li>
	<li>At this point, set the displayed vascular tension value to zero. Afterward, apply a 3 mN pull stimulus to the arterial ring by rotating the spiral axis of the bath.</li>
	<li>After incubation for 1 h in oxygen-saturated PSS buffer at 37 &#176;C, pH 7.40, set the tension value to 0 mN again on the tension control panel of the wire micrograph. The setting process of the initial tension of the arterial ring is shown in Figure 4.</li>
</ol>
5. Reactivity detection of coronary artery ring
<ol>
	<li>Perform the contractile activity of the coronary artery ring with the wire myograph technique14, and validate in three separate operations by stimulating with 60 mM of K+ solution for 10 min each.</li>
	<li>After each stimulation, flush the bath with oxygen-saturated PSS until vascular tone returns to its initial state. 
	NOTE: Only when the tension fluctuation of the three parallel measurements was less than 10%, and the amplitude of each contraction was greater than 1 mN/mm, qualified and highly active arterial rings could be used for further experiments. The activity verification of the rat coronary ring is shown in Figure 5.</li>
</ol>
6. Post-surgical treatment
<ol>
	<li>After surgery, euthanize the animals following institutionally approved protocols. 
	NOTE: For the present study, the animals were euthanized by inhaling excess isoflurane.</li>
</ol>

<ol><li>Malakar, A. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+review+on+coronary+artery+disease%2C+its+risk+factors%2C+and+therapeutics%5BTitle%5D)+AND+%22Journal+of+Cellular+Physiology%22%5BJournal%5D)">A review on coronary artery disease, its risk factors, and therapeutics.</a> Journal of Cellular Physiology. 234 (10), 16812-16823 (2019).</li>
<li>Murray, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((national+age-sex+specific+all-cause+and+cause-specific+mortality+for+240+causes+of+death%2C+1990-2013%3A+A+systematic+analysis+for+the+global+burden+of+disease+Study+2013%5BTitle%5D)+AND+%22The+Lancet%22%5BJournal%5D)">national age-sex specific all-cause and cause-specific mortality for 240 causes of death, 1990-2013: A systematic analysis for the global burden of disease Study 2013.</a> The Lancet. 385 (9963), 117-171 (2015).</li>
<li>Zhang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adenosine+and+adenosine+receptor-mediated+action+in+coronary+microcirculation%5BTitle%5D)+AND+%22Basic+Research+in+Cardiology%22%5BJournal%5D)">Adenosine and adenosine receptor-mediated action in coronary microcirculation.</a> Basic Research in Cardiology. 116 (1), 22(2021).</li>
<li>Allaqaband, H., Gutterman, D. D., Kadlec, A. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Physiological+consequences+of+coronary+arteriolar+dysfunction+and+its+influence+on+cardiovascular+disease%5BTitle%5D)+AND+%22Physiology%22%5BJournal%5D)">Physiological consequences of coronary arteriolar dysfunction and its influence on cardiovascular disease.</a> Physiology. 33 (5), 338-347 (2018).</li>
<li>Minelli, S., Minelli, P., Montinari, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Reflections+on+atherosclerosis%3A+Lesson+from+the+past+and+future+research+directions%5BTitle%5D)+AND+%22Journal+of+Multidisciplinary+Healthcare%22%5BJournal%5D)">Reflections on atherosclerosis: Lesson from the past and future research directions.</a> Journal of Multidisciplinary Healthcare. 13, 621-633 (2020).</li>
<li>Alvarez-Alvarez, M. M., Zanetti, D., Carreras-Torres, R., Moral, P., Athanasiadis, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+survey+of+sub-saharan+gene+flow+into+the+mediterranean+at+risk+loci+for+coronary+artery+disease%5BTitle%5D)+AND+%22European+Journal+of+Human+Genetics%22%5BJournal%5D)">A survey of sub-saharan gene flow into the mediterranean at risk loci for coronary artery disease.</a> European Journal of Human Genetics. 25 (4), 472-476 (2017).</li>
<li>LaCombe, P., Tariq, M. A., Lappin, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Physiology%2C+Afterload+Reduction%5BTitle%5D)+AND+%22StatPearls+%5BInternet%5D%22%5BJournal%5D)">Physiology, Afterload Reduction.</a> StatPearls [Internet]. , StatPearls Publishing. Treasure Island (FL). (2022).</li>
<li>Gutterman, D. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+human+microcirculation%3A+regulation+of+flow+and+beyond%5BTitle%5D)+AND+%22Circulation+Research%22%5BJournal%5D)">The human microcirculation: regulation of flow and beyond.</a> Circulation Research. 118 (1), 157-172 (2016).</li>
<li>Wang, G., Li, F., Hou, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Complementary+and+alternative+therapies+for+stable+angina+pectoris+of+coronary+heart+disease%3A+A+protocol+for+systematic+review+and+network+meta-analysis%5BTitle%5D)+AND+%22Medicine%22%5BJournal%5D)">Complementary and alternative therapies for stable angina pectoris of coronary heart disease: A protocol for systematic review and network meta-analysis.</a> Medicine. 101 (7), 28850(2022).</li>
<li>Markousis-Mavrogenis, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Coronary+microvascular+disease%3A+the+%26%2334%3Bmeeting+point%26%2334%3B+of+cardiology%5BTitle%5D)+AND+%22European+Journal+of+Clinical+Investigation%22%5BJournal%5D)">Coronary microvascular disease: the &#34;meeting point&#34; of cardiology.</a> European Journal of Clinical Investigation. 52 (5), 13737(2021).</li>
<li>Allison, B. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Fetal+in+vivo+continuous+cardiovascular+function+during+chronic+hypoxia%5BTitle%5D)+AND+%22The+Journal+of+Physiology%22%5BJournal%5D)">Fetal in vivo continuous cardiovascular function during chronic hypoxia.</a> The Journal of Physiology. 594 (5), 1247-1264 (2016).</li>
<li>Wenceslau, C. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Guidelines+for+the+measurement+of+vascular+function+and+structure+in+isolated+arteries+and+veins%5BTitle%5D)+AND+%22American+Journal+of+Physiology-Heart+and+Circulatory+Physiology%22%5BJournal%5D)">Guidelines for the measurement of vascular function and structure in isolated arteries and veins.</a> American Journal of Physiology-Heart and Circulatory Physiology. 321 (1), 77-111 (2021).</li>
<li>Liu, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Comparison+of+Ca2%2B+handling+for+the+regulation+of+vasoconstriction+between+rat+coronary+and+renal+arteries%5BTitle%5D)+AND+%22Journal+of+Vascular+Research%22%5BJournal%5D)">Comparison of Ca2+ handling for the regulation of vasoconstriction between rat coronary and renal arteries.</a> Journal of Vascular Research. 56 (4), 191-203 (2019).</li>
<li>Sun, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Isometric+contractility+measurement+of+the+mouse+mesenteric+artery+using+wire+myography%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Isometric contractility measurement of the mouse mesenteric artery using wire myography.</a> Journal of Visualized Experiments. (138), e58064(2018).</li>
<li>Guo, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Coronary+hypercontractility+to+acidosis+owes+to+the+greater+activity+of+TMEM16A%2FANO1+in+the+arterial+smooth+muscle+cells%5BTitle%5D)+AND+%22Biomedicine+%26+Pharmacotherapy%22%5BJournal%5D)">Coronary hypercontractility to acidosis owes to the greater activity of TMEM16A/ANO1 in the arterial smooth muscle cells.</a> Biomedicine & Pharmacotherapy. 139, 111615(2021).</li>
<li>Ping, N. N., Cao, L., Xiao, X., Li, S., Cao, Y. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+determination+of+optimal+initial+tension+in+rat+coronary+artery+using+wire+myography%5BTitle%5D)+AND+%22Physiological+Research%22%5BJournal%5D)">The determination of optimal initial tension in rat coronary artery using wire myography.</a> Physiological Research. 63 (1), 143-146 (2014).</li>
<li>Niccoli, G., Scalone, G., Lerman, A., Crea, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Coronary+microvascular+obstruction+in+acute+myocardial+infarction%5BTitle%5D)+AND+%22European+Heart+Journal%22%5BJournal%5D)">Coronary microvascular obstruction in acute myocardial infarction.</a> European Heart Journal. 37 (13), 1024-1033 (2016).</li>
<li>Mumma, B., Flacke, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Current+diagnostic+and+therapeutic+strategies+in+microvascular+angina%5BTitle%5D)+AND+%22Current+Emergency+and+Hospital+Medicine+Reports%22%5BJournal%5D)">Current diagnostic and therapeutic strategies in microvascular angina.</a> Current Emergency and Hospital Medicine Reports. 3 (1), 30-37 (2015).</li>
<li>Lanza, G. A., Parrinello, R., Figliozzi, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Management+of+microvascular+angina+pectoris%5BTitle%5D)+AND+%22American+Journal+of+Cardiovascular+Drugs%22%5BJournal%5D)">Management of microvascular angina pectoris.</a> American Journal of Cardiovascular Drugs. 14 (1), 31-40 (2014).</li>
<li>Zhu, T. Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Beneficial+effects+of+intracoronary+tirofiban+bolus+administration+following+upstream+intravenous+treatment+in+patients+with+ST-elevation+myocardial+infarction+undergoing+primary+percutaneous+coronary+intervention%3A+The+ICT-AMI+study%5BTitle%5D)+AND+%22International+Journal+of+Cardiology%22%5BJournal%5D)">Beneficial effects of intracoronary tirofiban bolus administration following upstream intravenous treatment in patients with ST-elevation myocardial infarction undergoing primary percutaneous coronary intervention: The ICT-AMI study.</a> International Journal of Cardiology. 165 (3), 437-443 (2013).</li>
<li>Huang, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Restoration+of+coronary+flow+in+patients+with+no-reflow+after+primary+coronary+intervention+of+acute+myocardial+infarction+%28RECOVER%29%5BTitle%5D)+AND+%22American+Heart+Journal%22%5BJournal%5D)">Restoration of coronary flow in patients with no-reflow after primary coronary intervention of acute myocardial infarction (RECOVER).</a> American Heart Journal. 164 (3), 394-401 (2012).</li>
<li>Fu, W. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Anti-atherosclerosis+and+cardio-protective+effects+of+the+Angong+Niuhuang+Pill+on+a+high+fat+and+vitamin+D3+induced+rodent+model+of+atherosclerosis%5BTitle%5D)+AND+%22Journal+of+Ethnopharmacology%22%5BJournal%5D)">Anti-atherosclerosis and cardio-protective effects of the Angong Niuhuang Pill on a high fat and vitamin D3 induced rodent model of atherosclerosis.</a> Journal of Ethnopharmacology. 195, 118-126 (2017).</li>
<li>Li, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Chinese+medicine+She-Xiang-Xin-Tong-Ning%2C+containing+moschus%2C+corydalis+and+ginseng%2C+protects+from+myocardial+ischemia+injury+via+angiogenesis%5BTitle%5D)+AND+%22The+American+Journal+of+Chinese+Medicine%22%5BJournal%5D)">Chinese medicine She-Xiang-Xin-Tong-Ning, containing moschus, corydalis and ginseng, protects from myocardial ischemia injury via angiogenesis.</a> The American Journal of Chinese Medicine. 48 (1), 107-126 (2020).</li>
<li>Wu, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Three+dimensional+reconstruction+of+coronary+artery+stents+from+optical+coherence+tomography%3A+Experimental+validation+and+clinical+feasibility%5BTitle%5D)+AND+%22Scientific+Reports%22%5BJournal%5D)">Three dimensional reconstruction of coronary artery stents from optical coherence tomography: Experimental validation and clinical feasibility.</a> Scientific Reports. 11 (1), 1-15 (2021).</li>
<li>Liu, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Janus-like+role+of+fibroblast+growth+factor+2+in+arteriosclerotic+coronary+artery+disease%3A+Atherogenesis+and+angiogenesis%5BTitle%5D)+AND+%22Atherosclerosis%22%5BJournal%5D)">Janus-like role of fibroblast growth factor 2 in arteriosclerotic coronary artery disease: Atherogenesis and angiogenesis.</a> Atherosclerosis. 229 (1), 10-17 (2013).</li>
<li>Hu, G., Li, X., Zhang, S., Wang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Association+of+rat+thoracic+aorta+dilatation+by+astragaloside+IV+with+the+generation+of+endothelium-derived+hyperpolarizing+factors+and+nitric+oxide%2C+and+the+blockade+of+Ca2%2B+channels%5BTitle%5D)+AND+%22Biomedical+reports%22%5BJournal%5D)">Association of rat thoracic aorta dilatation by astragaloside IV with the generation of endothelium-derived hyperpolarizing factors and nitric oxide, and the blockade of Ca2+ channels.</a> Biomedical reports. 5 (1), 27-34 (2016).</li>
<li>Guo, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Anticonstriction+effect+of+MCA+in+rats+by+danggui+buxue+decoction%5BTitle%5D)+AND+%22Frontiers+in+Pharmacology%22%5BJournal%5D)">Anticonstriction effect of MCA in rats by danggui buxue decoction.</a> Frontiers in Pharmacology. 12, 749915(2021).</li>
<li>Jing, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Apigenin+relaxes+rat+intrarenal+arteries%2C+depresses+Ca2%2B-activated+Cl%E2%88%92+currents+and+augments+voltage-dependent+K%2B+currents+of+the+arterial+smooth+muscle+cells%5BTitle%5D)+AND+%22Biomedicine+%26+Pharmacotherapy%22%5BJournal%5D)">Apigenin relaxes rat intrarenal arteries, depresses Ca2+-activated Cl− currents and augments voltage-dependent K+ currents of the arterial smooth muscle cells.</a> Biomedicine & Pharmacotherapy. 115, 108926(2019).</li>
</ol>

The authors have nothing to disclose.

The disturbance of coronary microcirculation, which involves a wide range of patients with CAD, has been gradually recognized and concerned the basis for adequate myocardium perfusion. Considering the serious complications of sudden coronary heart disease and cardiovascular disease, timely drug prevention and treatment are extremely important for a clinical individual with CAD17. Inevitably, the secrecy of coronary artery anatomy and the complexity of its physiological structure have severely rest...

Coronary artery disease (CAD) has been widely recognized and concerned as a typical and representative cardiovascular disease, being the leading cause of death in both developed and developing countries1,2. As a blood and oxygen supply route for normal cardiac physiological function, circulating blood enters and nourishes the heart through two main coronary arteries and a blood vascular network on the surface of the myocardium3,4. Cholesterol and fat deposits in the coronary arteries cut off the heart&#39;s blood supply and the violent inflammatory response of the vascular system, causing atherosclerosis, stable angina, unstable angina, myocardial infarction, or sudden cardiac death5,6. In response to pathological stenosis of the coronary arteries, compensatory accelerated physiological heartbeat satisfies the blood supply of the heart itself or vital organs of the body by increasing the output of the left ventricle7. If prolonged coronary stenosis is not relieved in time, extensive new blood vessels may develop in certain areas of the heart8. At present, the clinical treatment of CAD often adopts drug thrombolysis or surgical mechanical thrombolysis and an exogenous bionic vascular bypass with frequent medication and great surgical disability9. Therefore, the functional investigation of coronary artery physiological activity is still an urgent breakthrough for cardiovascular diseases10.
There are no available technical means to detect coronary physiological activity, except for wireless telemetry systems, which can dynamically record in vivo coronary pressure, vascular tension, blood oxygen saturation, and pH values11. Therefore, considering coronary arteries&#39; textural secrecy and complexity, accurate identification and isolation of coronary arteries are undoubtedly the best choices for exploring multiple mechanisms of CAD in vitro4.
A series multi myograph system, in particular a wire micrograph microvascular tension detector (see Table of Materials), is a very mature marketable device for recording in vitro tissue tension changes of small vascular, lymphatic, and bronchial tubes with the characteristics of high precision and continuous dynamic recording12. The said system has been extensively employed to record in vitro tissue tension characteristics of cavity structures with diameters of 60 &#181;m to 10 mm. The continuous heating features of the platform of the wire micrograph largely offset the stimulation of the adverse external environment. Meanwhile, the constant inputs of the gas mixture and the pH values allow us to obtain more accurate vascular tension data in a similar physiological state13. However, considering the complexity of anatomical localization of rat coronary arteries (Figure 1), its isolation has been perplexing and limiting the mechanism&#39;s exploration of diversified cardiovascular disease and drug development. Therefore, the present protocol introduces the anatomical location and separation process of the rat coronary artery in detail, followed by tension measurement on the platform of the wire micrograph14.

<table><tbody><tr><td>Apigenin</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>150731</td><td></td></tr><tr><td>CaCl&lt;sub&gt;2&lt;/sub&gt;</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>A501330</td><td></td></tr><tr><td>D-glucose</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>A610219</td><td></td></tr><tr><td>HEPES</td><td>Xiya Reagent Co., Ltd., Shandong, China</td><td>S3872</td><td></td></tr><tr><td>KCl</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>A100395</td><td></td></tr><tr><td>KH&lt;sub&gt;2&lt;/sub&gt;PO&lt;sub&gt;4&lt;/sub&gt;</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>A100781</td><td></td></tr><tr><td>LabChart Professional version 8.3&amp;nbsp;</td><td>ADInstruments, Australia</td><td>&amp;mdash;</td><td></td></tr><tr><td>MgCl&lt;sub&gt;2&lt;/sub&gt;&amp;middot;6H&lt;sub&gt;2&lt;/sub&gt;O</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>A100288</td><td></td></tr><tr><td>Multi myograph system&amp;nbsp;</td><td>Danish Myo Technology, Aarhus, Denmark</td><td>620M</td><td></td></tr><tr><td>NaCl</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>A100241</td><td></td></tr><tr><td>NaHCO&lt;sub&gt;3&lt;/sub&gt;</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>A100865</td><td></td></tr><tr><td>Steel wires</td><td>Danish Myo Technology, Aarhus, Denmark</td><td>400447</td><td></td></tr><tr><td>U46619</td><td>Sigma, USA</td><td>D8174</td><td></td></tr></tbody></table>

standardized rat coronary ring preparation and real-time recording of dynamic tension changes along vessel diameter

Standardized Rat Coronary Ring Preparation and Real-Time Recording of Dynamic Tension Changes Along Vessel Diameter

As a key event of cardiovascular system diseases, coronary artery disease (CAD) has been widely regarded as the main culprit of atherosclerosis, myocardial infarction, and angina pectoris, which seriously threaten the life and health of people all over the world. However, how to record the dynamic biomechanical characteristics of isolated blood vessels has long puzzled people. Meanwhile, precise positioning and isolation of coronary arteries to measure in vitro dynamic vascular tension changes have become a trend in CAD drug development. The present protocol describes the macroscopic identification and microscopic separation of rat coronary arteries. The contraction and dilation function of the coronary artery ring along the vessel diameter was monitored using the established multi myograph system. The standardized and programmed protocols of coronary ring tension measurement, from sampling to data acquisition, tremendously improve the repeatability of the experimental data, which ensures the authenticity of vascular tension records after physiological, pathological, and drug intervention.

Anatomically positioned, rat coronary arteries distributed and hidden deep in myocardial tissue were not easily recognized. By comparing the coronary arteries of humans (Figure 1A) and rats (Figure 1B), rapid and accurate separation of rat coronary arteries was conducted according to the sampling process in Figure 2. After precisely locating the right auricle, pulmonary artery, and apex from the front under an optical microscope, th...

The present protocol describes the wire myograph technique for measuring vascular reactivity of the rat coronary artery.

As a key event of cardiovascular system diseases, coronary artery disease (CAD) has been widely regarded as the main culprit of atherosclerosis, ...

standardized-rat-coronary-ring-preparation-real-time-recording

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Evaluation of Vascular Control Mechanisms Utilizing Video Microscopy of Isolated Resistance Arteries of Rats

The protocol describes the use of wire myography to evaluate the transmural isometric tension of mesenteric arteries isolated from mice, with special consideration of the modulation by factors released from endothelial cells and perivascular adipose tissues.

Assessment of Vascular Tone Responsiveness using Isolated Mesenteric Arteries with a Focus on Modulation by Perivascular Adipose Tissues

Presented here is a protocol to study the coronary microcirculation in living murine heart tissue by ex vivo monitoring of the arterial perfusion pressure and flow that maintains the pressure, as well as vascular tree components including the capillary beds and pericytes, as the septal artery is cannulated and pressurized.

Dynamic Measurement and Imaging of Capillaries, Arterioles, and Pericytes in Mouse Heart

The present protocol describes the concepts and technical application of the tensometric myograph technique using a multi-chamber myograph system in the experimental ex vivo assessment of mouse aortic endothelial function.

Measurement of Endothelium-Dependent Vasorelaxation in the Mouse Thoracic Aorta Using Tensometric Small Volume Chamber Myography

This protocol describes a method to simultaneously measure cytosolic, free calcium [Ca2+]i and vessel diameter in contracting lymph vessels in real time and then calculate absolute Ca2+ concentrations as well as contractility/rhythmicity parameters. This protocol can be used to study Ca2+ and contractile dynamics across a variety of experimental conditions.

Real-time Evaluation of Absolute, Cytosolic, Free Ca2+ and Corresponding Contractility in Isolated, Pressurized Lymph Vessels

Right heart failure (RHF) is characterized by right-sided cardiac dilation and hypertrophy, leading to ventricular and atrial malfunction. Cardiopulmonary conditions associated with RHF are accompanied by an increased risk for cardiac arrhythmias. This article describes a standardized model of pulmonary artery banding-induced RHF associated with enhanced ventricular and atrial arrhythmogenesis.

Rat Model of Right-Sided Cardiac Remodeling and Arrhythmia Using Pulmonary Artery Banding

This article presents a step-by-step protocol for the isolation and functional evaluation of human renal arterial branches, facilitating preclinical studies for pharmaceutical development.

Multilevel Microdissection and Functional-Structural Profiling of Human Renal Arterial Branches

We are developing a dynamic adaptive exposure technique using our scanning beam digital X-ray system. Rather than exposing an object uniformly, the exposure is adapted depending on the opacity of the object. Here we show an experiment on an anthropomorphic phantom that resulted in a dose saving of 30%.

X-ray Dose Reduction through Adaptive Exposure in Fluoroscopic Imaging

Visualization of the coronary vessels is critical to advancing our understanding of cardiovascular diseases. Here we describe a method for perfusing murine coronary vasculature with a radiopaque silicone rubber (Microfil), in preparation for micro-Computed Tomography (μCT) imaging.

Retrograde Perfusion and Filling of Mouse Coronary Vasculature as Preparation for Micro Computed Tomography Imaging

Learn the wire myograph technique for measuring vascular reactivity in rat coronary arteries. This standardized method enables stable recording of dynamic tension changes in vessels from 60 micrometers to 10 millimeters.

Standardized Rat Coronary Ring Preparation and Real-Time Recording of Dynamic Tension Changes Along Vessel Diameter

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