Name: assessment of vascular tone responsiveness using isolated mesenteric arteries with a focus on modulation by perivascular adipose tissues
Uploaded: 2019-06-03T15:00:00
Description: Watch this Scientific Journal Video about Assessment of Vascular Tone Responsiveness using Isolated Mesenteric Arteries with a Focus on Modulation by Perivascular Adipose Tissues at JoVE.com

This work was financially support by the grants from Research Grant Council of Hong Kong [17124718 and 17121714], Hong Kong Health and Medical Research Fund [13142651 and 13142641], Collaborative Research Fund of Hong Kong [C7055-14G], and the National Basic Research Program of China [973 Program 2015CB553603].

All animals used for the following study were provided by the Laboratory Animal Unit of the Faculty of Medicine, The University of Hong Kong. Ethical approval was obtained from the departmental Committee on Use of Laboratory Animals for Teaching and Research (CULATR, no.: 4085-16).
1. Preparations
<ol>
	<li>Preparation of drugs
	<ol>
		<li>Store drugs appropriately as stated in the Material Safety Data Sheet (MSDS) immediately after receiving them. Dissolve the drugs in powd...

<ol>
	<li>Furchgott, R. F., Zawadzki, J. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+obligatory+role+of+endothelial+cells+in+the+relaxation+of+arterial+smooth+muscle+by+acetylcholine.">The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine.</a> Nature. 288 (5789), 373-376 (1980).</li><li>Furchgott, R. F., Vanhoutte, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelium-derived+relaxing+and+contracting+factors.">Endothelium-derived relaxing and contracting factors.</a> The FASEB Journal. 3 (9), 2007-2018 (1989).</li><li>Feletou, M., Kohler, R., Vanhoutte, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelium-derived+vasoactive+factors+and+hypertension:+possible+roles+in+pathogenesis+and+as+treatment+targets.">Endothelium-derived vasoactive factors and hypertension: possible roles in pathogenesis and as treatment targets.</a> Current Hypertension Reports. 12 (4), 267-275 (2010).</li><li>Vanhoutte, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+dysfunction:+the+first+step+toward+coronary+arteriosclerosis.">Endothelial dysfunction: the first step toward coronary arteriosclerosis.</a> Circulation Journal. 73 (4), 595-601 (2009).</li><li>Feletou, M., Huang, Y., Vanhoutte, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelium-mediated+control+of+vascular+tone:+COX-1+and+COX-2+products.">Endothelium-mediated control of vascular tone: COX-1 and COX-2 products.</a> British Journal of Pharmacology. 164 (3), 894-912 (2011).</li><li>Harrison, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+and+molecular+mechanisms+of+endothelial+cell+dysfunction.">Cellular and molecular mechanisms of endothelial cell dysfunction.</a> Journal of Clinical Investigation. 100 (9), 2153 (1997).</li><li>Vanhoutte, P. M., Shimokawa, H., Tang, E. H., Feletou, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+dysfunction+and+vascular+disease.">Endothelial dysfunction and vascular disease.</a> Acta physiologica. 196 (2), 193-222 (2009).</li><li>Kl&#246;&#223;, S., Bouloumi&#233;, A., M&#252;lsch, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aging+and+chronic+hypertension+decrease+expression+of+rat+aortic+soluble+guanylyl+cyclase.">Aging and chronic hypertension decrease expression of rat aortic soluble guanylyl cyclase.</a> Hypertension. 35 (1), 43-47 (2000).</li><li>Csiszar, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aging-induced+phenotypic+changes+and+oxidative+stress+impair+coronary+arteriolar+function.">Aging-induced phenotypic changes and oxidative stress impair coronary arteriolar function.</a> Circulation Research. 90 (11), 1159-1166 (2002).</li><li>Guo, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+SIRT1+prevents+age-induced+impairment+of+vasodilator+responses+by+enhancing+the+expression+and+activity+of+soluble+guanylyl+cyclase+in+smooth+muscle+cells.">Endothelial SIRT1 prevents age-induced impairment of vasodilator responses by enhancing the expression and activity of soluble guanylyl cyclase in smooth muscle cells.</a> Cardiovascular Research. , (2018).</li><li>Auch-Schwelk, W., Katusic, Z. S., Vanhoutte, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nitric+oxide+inactivates+endothelium-derived+contracting+factor+in+the+rat+aorta.">Nitric oxide inactivates endothelium-derived contracting factor in the rat aorta.</a> Hypertension. 19 (5), 442-445 (1992).</li><li>Tang, E. H., Feletou, M., Huang, Y., Man, R. Y., Vanhoutte, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acetylcholine+and+sodium+nitroprusside+cause+long-term+inhibition+of+EDCF-mediated+contractions.">Acetylcholine and sodium nitroprusside cause long-term inhibition of EDCF-mediated contractions.</a> American Journal of Physiology - Heart and Circulation Physiology. 289 (6), H2434-H2440 (2005).</li><li>Ghiadoni, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+function+and+common+carotid+artery+wall+thickening+in+patients+with+essential+hypertension.">Endothelial function and common carotid artery wall thickening in patients with essential hypertension.</a> Hypertension. 32 (1), 25-32 (1998).</li><li>Xu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age-related+Impairment+of+Vascular+Structure+and+Functions.">Age-related Impairment of Vascular Structure and Functions.</a> Aging and Disease. 8 (5), 590-610 (2017).</li><li>Tabit, C. E., Chung, W. B., Hamburg, N. M., Vita, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+dysfunction+in+diabetes+mellitus:+Molecular+mechanisms+and+clinical+implications.">Endothelial dysfunction in diabetes mellitus: Molecular mechanisms and clinical implications.</a> Reviews in Endocrine &amp; Metabolic Disorders. 11 (1), 61-74 (2010).</li><li>Tanaka, K., Sata, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Roles+of+perivascular+adipose+tissue+in+the+pathogenesis+of+atherosclerosis.">Roles of perivascular adipose tissue in the pathogenesis of atherosclerosis.</a> Frontiers in Physiology. 9, 3 (2018).</li><li>Brown, N. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perivascular+adipose+tissue+in+vascular+function+and+disease:+a+review+of+current+research+and+animal+models.">Perivascular adipose tissue in vascular function and disease: a review of current research and animal models.</a> Arteriosclerosis Thrombosis and Vascular Biology. 34 (8), 1621-1630 (2014).</li><li>Lohn, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Periadventitial+fat+releases+a+vascular+relaxing+factor.">Periadventitial fat releases a vascular relaxing factor.</a> The FASEB Journal. 16 (9), 1057-1063 (2002).</li><li>G&#225;lvez-Prieto, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+reduction+in+the+amount+and+anti-contractile+effect+of+periadventitial+mesenteric+adipose+tissue+precedes+hypertension+development+in+spontaneously+hypertensive+rats.">A reduction in the amount and anti-contractile effect of periadventitial mesenteric adipose tissue precedes hypertension development in spontaneously hypertensive rats.</a> Hypertension research. 31 (7), 1415 (2008).</li><li>Gao, Y. J., Lu, C., Su, L. Y., Sharma, A., Lee, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+vascular+function+by+perivascular+adipose+tissue:+the+role+of+endothelium+and+hydrogen+peroxide.">Modulation of vascular function by perivascular adipose tissue: the role of endothelium and hydrogen peroxide.</a> British Journal of Pharmacology. 151 (3), 323-331 (2007).</li><li>Gao, Y. -. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perivascular+adipose+tissue+promotes+vasoconstriction:+the+role+of+superoxide+anion.">Perivascular adipose tissue promotes vasoconstriction: the role of superoxide anion.</a> Cardiovascular Research. 71 (2), 363-373 (2006).</li><li>Szasz, T., Webb, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perivascular+adipose+tissue:+more+than+just+structural+support.">Perivascular adipose tissue: more than just structural support.</a> Clinical Science (London). 122 (1), 1-12 (2012).</li><li>Ramirez, J. G., O'Malley, E. J., Ho, W. S. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pro-contractile+effects+of+perivascular+fat+in+health+and+disease.">Pro-contractile effects of perivascular fat in health and disease.</a> Brish Journal of Pharmacology. 174 (20), 3482-3495 (2017).</li><li>Hajer, G. R., van Haeften, T. W., Visseren, F. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adipose+tissue+dysfunction+in+obesity,+diabetes,+and+vascular+diseases.">Adipose tissue dysfunction in obesity, diabetes, and vascular diseases.</a> European Heart Journal. 29 (24), 2959-2971 (2008).</li><li>Mulvany, M. J., Halpern, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contractile+properties+of+small+arterial+resistance+vessels+in+spontaneously+hypertensive+and+normotensive+rats.">Contractile properties of small arterial resistance vessels in spontaneously hypertensive and normotensive rats.</a> Circulation Research. 41 (1), 19-26 (1977).</li><li>Mulvany, M. J., Halpern, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+properties+of+vascular+smooth+muscle+cells+in+situ.">Mechanical properties of vascular smooth muscle cells in situ.</a> Nature. 260 (5552), 617-619 (1976).</li><li>del Campo, L., Ferrer, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wire+myography+to+study+vascular+tone+and+vascular+structure+of+isolated+mouse+arteries.">Wire myography to study vascular tone and vascular structure of isolated mouse arteries.</a> Methods in Molecular Biology. 1339, 255-276 (2015).</li><li>Dobrin, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+initial+length+on+length-tension+relationship+of+vascular+smooth+muscle.">Influence of initial length on length-tension relationship of vascular smooth muscle.</a> American Journal of Physiology. 225 (3), 664-670 (1973).</li><li>Xu, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calorie+restriction+prevents+metabolic+aging+caused+by+abnormal+SIRT1+function+in+adipose+tissues.">Calorie restriction prevents metabolic aging caused by abnormal SIRT1 function in adipose tissues.</a> Diabetes. 64 (5), 1576-1590 (2015).</li><li>Sheykhzade, M., Nyborg, N. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Caliber+dependent+calcitonin+gene-related+peptide-induced+relaxation+in+rat+coronary+arteries:+effect+of+K++on+the+tachyphylaxis.">Caliber dependent calcitonin gene-related peptide-induced relaxation in rat coronary arteries: effect of K+ on the tachyphylaxis.</a> European Journal of Pharmacology. 351 (1), 53-59 (1998).</li><li>Soltis, E. E., Cassis, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+perivascular+adipose+tissue+on+rat+aortic+smooth+muscle+responsiveness.">Influence of perivascular adipose tissue on rat aortic smooth muscle responsiveness.</a> Clinical and Experimental Hypertension A. 13 (2), 277-296 (1991).</li><li>Lohn, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Periadventitial+fat+releases+a+vascular+relaxing+factor.">Periadventitial fat releases a vascular relaxing factor.</a> FASEB Journal. 16 (9), 1057-1063 (2002).</li><li>Fesus, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adiponectin+is+a+novel+humoral+vasodilator.">Adiponectin is a novel humoral vasodilator.</a> Cardiovascular Research. 75 (4), 719-727 (2007).</li><li>Greenstein, A. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Local+inflammation+and+hypoxia+abolish+the+protective+anticontractile+properties+of+perivascular+fat+in+obese+patients.">Local inflammation and hypoxia abolish the protective anticontractile properties of perivascular fat in obese patients.</a> Circulation. 119 (12), 1661-1670 (2009).</li><li>Yudkin, J. S., Eringa, E., Stehouwer, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term="Vasocrine"+signalling+from+perivascular+fat:+a+mechanism+linking+insulin+resistance+to+vascular+disease.">"Vasocrine" signalling from perivascular fat: a mechanism linking insulin resistance to vascular disease.</a> Lancet. 365 (9473), 1817-1820 (2005).</li><li>Xia, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uncoupling+of+endothelial+nitric+oxide+synthase+in+perivascular+adipose+tissue+of+diet-induced+obese+mice.">Uncoupling of endothelial nitric oxide synthase in perivascular adipose tissue of diet-induced obese mice.</a> Arteriosclerosis Thrombosis and Vascular Biology. 36 (1), 78-85 (2016).</li><li>Xia, N., Forstermann, U., Li, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+resveratrol+on+eNOS+in+the+endothelium+and+the+perivascular+adipose+tissue.">Effects of resveratrol on eNOS in the endothelium and the perivascular adipose tissue.</a> Annals of the New York Academy of Sciences. 1403 (1), 132-141 (2017).</li><li>Schinzari, F., Tesauro, M., Cardillo, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+and+perivascular+adipose+tissue+abnormalities+in+obesity-related+vascular+dysfunction:+novel+targets+for+treatment.">Endothelial and perivascular adipose tissue abnormalities in obesity-related vascular dysfunction: novel targets for treatment.</a> Journal of Cardiovascular Pharmacology. 69 (6), 360-368 (2017).</li><li>Liu, J. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lipocalin-2+deficiency+prevents+endothelial+dysfunction+associated+with+dietary+obesity:+role+of+cytochrome+P450+2C+inhibition.">Lipocalin-2 deficiency prevents endothelial dysfunction associated with dietary obesity: role of cytochrome P450 2C inhibition.</a> British Journal of Pharmacology. 165 (2), 520-531 (2012).</li><li>Martinez-Quinones, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypertension+induced+morphological+and+physiological+changes+in+cells+of+the+arterial+wall.">Hypertension induced morphological and physiological changes in cells of the arterial wall.</a> American Journal of Hypertension. 31 (10), 1067-1078 (2018).</li><li>Outzen, E. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Translational+value+of+mechanical+and+vasomotor+properties+of+mouse+isolated+mesenteric+resistance-sized+arteries.">Translational value of mechanical and vasomotor properties of mouse isolated mesenteric resistance-sized arteries.</a> Pharmacology Research and Perspectives. 3 (6), e00200 (2015).</li><li>Sheykhzade, M., Simonsen, A. H., Boonen, H. C., Outzen, E. M., Nyborg, N. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+ageing+on+the+passive+and+active+tension+and+pharmacodynamic+characteristics+of+rat+coronary+arteries:+age-dependent+increase+in+sensitivity+to+5-HT+and+K+.">Effect of ageing on the passive and active tension and pharmacodynamic characteristics of rat coronary arteries: age-dependent increase in sensitivity to 5-HT and K+.</a> Pharmacology. 90 (3-4), 160-168 (2012).</li></ol>

Apart from the endothelial cells, signals derived from PVAT play an important role in the regulation of smooth muscle tone reactivity30. Healthy PVAT releases NO and anti-inflammatory adiponectin to exert an anti-contractile effect on arteries, which is lost under pathological conditions such as obesity and metabolic syndrome31,32. In disease states, PVAT contributes to the development of endothelial dysfunction and other cardiovascular ab...

Dilatations and constrictions of arteries are achieved by relaxations and contractions, respectively, of their vascular smooth muscle cells. Changes in vascular responsiveness of small arteries contribute to the homeostatic regulation of arterial blood pressure by autonomic nerves and hormones present in the blood (e.g., catecholamines, angiotensin II, serotonin, vasopressin). At the local level, the vascular responses of smooth muscle cells are modulated by signals from both the endothelial cells of the intima and the adipose tissue surrounding the arteries (Figure 1).
The endothelium is not only a passive barrier, but also serves as a surface to exchange signals between the blood and the underlying vascular smooth muscle cells. By releasing various vasoactive substances, the endothelium plays a critical role in the local control of vascular tone responses1. For example, in response to acetylcholine, endothelial nitric oxide synthase (eNOS) is activated in the endothelium to produce nitric oxide (NO), which induces relaxation of the underlying vascular smooth muscle by activating soluble guanylyl cyclase (sGC)2. Other vasoactive substances include the products of cyclooxygenases (e.g., prostacyclin and thromboxane A2), lipoxygenase (e.g., 12-hydroxyeicosatetraenoic acids, 12-HETE), and cytochrome P450 monooxygenases (HETEs and epoxyeicosatrienoic acids, EETs), reactive oxygen species (ROS), and vasoactive peptides (e.g., endothelin-1 and angiotensin II), and endothelium-derived hyperpolarizing factors (EDHF)3. A delicate balance between endothelium-derived vasodilators and vasoconstrictors maintain the local vasomotor tone4,5.
Endothelial dysfunction is characterized by the impairment in endothelium-dependent vasodilatation6, a hallmark of vascular aging7. With age, the ability of endothelium to promote vasodilatation is progressively reduced, due largely to a decreased NO bioavailability, as well as the abnormal expression and function of eNOS in the endothelium and sGC in the vascular smooth muscle cells8,9,10. Reduced NO bioavailability potentiates the production of endothelium-dependent vasoconstrictors11,12. In aged arteries, endothelial dysfunction causes hyperplasia in the media, as reflected by the marked increases in wall thickness, number of medial nuclei, which are reminiscent of the arterial thickening in hypertension and atherosclerosis observed in human patients13,14. In addition, pathophysiological conditions such as obesity, diabetes or hypertension accelerate the development of endothelial dysfunction15,16.
Perivascular adipose tissue (PVAT) releases numerous adipokines to regulate vascular structure and function17. The anti-contractile effect of PVAT is mediated by relaxing factors, such as adiponectin, NO, hydrogen peroxide and hydrogen sulphide18,19,20. However, depending on the location and pathophysiological condition, PVAT also can enhance contractile responses&#160;in various arteries21. The pro-contractile substances produced by PVAT include angiotensin-II, leptin, resistin, and ROS22,23.&#160; In most of the studies on isolated blood vessels, PVAT has been considered as a simple structural support for the vasculature and thus removed during the preparation of blood vessel ring segments. Since adipose dysfunction represents an independent risk factor for hypertension and associated cardiovascular complications24, the PVAT surrounding the blood vessels should be considered when investigating the vascular responsiveness of different arteries.
The multi wire myograph systems have been widely used to investigate the vasomotor functions of a variety of blood vessels, including the aorta, mesenteric, renal, femoral, cerebral and coronary arteries25,26. The protocols described herein will use wire myography to evaluate vascular responsiveness in mesenteric arteries isolated from genetically modified mouse models, with a special focus on the modulation by PVAT.

<table>
	<tbody>
		<tr>
			<td>Acetylcholine</td>
			<td>Sigma-Aldrich</td>
			<td>A6625</td>
			<td>Stock concentration: 10-1 M 
			Working concentration: 10-10 to 10-5 M</td>
		</tr>
		<tr>
			<td>L-NAME (N&#969;-nitro-L-arginine methyl ester)</td>
			<td>Sigma-Aldrich</td>
			<td>N5751</td>
			<td>Stock concentration: 3 x 10-2 M 
			Working concentration: 10-4 M</td>
		</tr>
		<tr>
			<td>Phenylephrine</td>
			<td>Sigma-Aldrich</td>
			<td>P6126</td>
			<td>Stock concentration: 10-2 M 
			Working concentration: 10-10 to 10-5 M</td>
		</tr>
		<tr>
			<td>U46619 (9,11-dideoxy-9&#945;,11&#945;methanoepoxy prostaglandin F2&#945;)</td>
			<td>Enzo</td>
			<td>BML-PG023-0001</td>
			<td>Stock concentration: 10-5 M 
			Working concentration: 1-3 x 10-8 M</td>
		</tr>
		<tr>
			<td>Multiwire myograph</td>
			<td>Danish MyoTechnology (DMT)</td>
			<td>620M</td>
			<td></td>
		</tr>
		<tr>
			<td>PowerLab 4/26</td>
			<td>ADInstruments</td>
			<td>ML848</td>
			<td></td>
		</tr>
		<tr>
			<td>Labchart7</td>
			<td>ADInstruments</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>Adipo-SIRT1 wild type mice</td>
			<td>Laboratory Animal Unit, The University of Hong Kong</td>
			<td>CULATR NO.: 4085-16</td>
			<td></td>
		</tr>
		<tr>
			<td>Silicon-coated Petri dishes</td>
			<td>Danish MyoTechnology (DMT)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Tungsten wires</td>
			<td>Danish MyoTechnology (DMT)</td>
			<td>300331</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical tools</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

assessment of vascular tone responsiveness using isolated mesenteric arteries with a focus on modulation by perivascular adipose tissues

Altered vascular tone responsiveness to pathophysiological stimuli contributes to the development of a wide range of cardiovascular and metabolic diseases. Endothelial dysfunction represents a major culprit for the reduced vasodilatation and enhanced vasoconstriction of arteries. Adipose (fat) tissues surrounding the arteries play important roles in the regulation of endothelium-dependent relaxation and/or contraction of the vascular smooth muscle cells. The cross-talks between the endothelium and perivascular adipose tissues can be assessed ex vivo using mounted blood vessels by a wire myography system. However, optimal settings should be established for arteries derived from animals of different species, ages, genetic backgrounds and/or pathophysiological conditions.

Examination of the length/tension relationships to obtain the normalization factor k 
The amount of stretch applied to a vessel segment influences the extent of the actin-myosin interaction and hence the maximal active force developed. Thus, for every type of blood vessel, determining the amount of stretch needed for maximal active force is required for proper myography studies. Here, normalization of t...

Watch this Scientific Journal Video about Assessment of Vascular Tone Responsiveness using Isolated Mesenteric Arteries with a Focus on Modulation by Perivascular Adipose Tissues at JoVE.com

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

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.

Altered vascular tone responsiveness to pathophysiological stimuli contributes to the development of a wide range of cardiovascular and metabolic ...

Research

JoVE Journal

Biology

Right Hemihepatectomy by Suprahilar Intrahepatic Transection of the Right Hemipedicle using a Vascular Stapler

Successful hepatic resection requires profound anatomical knowledge and delicate surgical technique. Hemihepatectomies are mostly performed after ...

The ex vivo Isolated Skeletal Microvessel Preparation for Investigation of Vascular Reactivity

The ex vivo Isolated Skeletal Microvessel Preparation for Investigation of Vascular Reactivity

The isolated  microvessel preparation is an ex vivo preparation that allows for examination of the different contributions of  factors that control vessel ...

Using Digital Image Correlation to Characterize Local Strains on Vascular Tissue Specimens

Characterization of the mechanical behavior of biological and engineered soft tissues is a central component of fundamental biomedical research and ...

Analysis of Cell Suspensions Isolated from Solid Tissues by Spectral Flow Cytometry

Flow cytometry has been used for the past 40 years to define and analyze the phenotype of lymphoid and other hematopoietic cells. Initially restricted to ...

Differentiation Capacity of Human Aortic Perivascular Adipose Progenitor Cells

Adipose tissue is a rich source of multi-potent mesenchymal stem cells (MSC) capable of differentiating into osteogenic, adipogenic, and chondrogenic ...

Preparation of Adipose Progenitor Cells from Mouse Epididymal Adipose Tissues

Obesity and metabolic disorders such as diabetes, heart disease, and cancer, are all associated with dramatic adipose tissue remodeling. Tissue-resident ...

Visualization and Quantification of Brown and Beige Adipose Tissues in Mice using [18F]FDG Micro-PET/MR Imaging

Visualization and Quantification of Brown and Beige Adipose Tissues in Mice using [18F]FDG Micro-PET/MR Imaging

Brown and beige adipocytes are now recognized as potential therapeutic targets for obesity and metabolic syndromes. Non-invasive molecular imaging methods ...

Mouse Abdominal Aortic Aneurysm Model Induced by Perivascular Application of Elastase

Abdominal aortic aneurysm (AAA), although primarily asymptomatic, is potentially life-threatening as the rupture of AAA usually has a devastating outcome. ...

Semi-Automated Isolation of Murine White Adipose Tissue

Semi-Automated Isolation of the Stromal Vascular Fraction from Murine White Adipose Tissue Using a Tissue Dissociator

Semi-Automated Isolation of the Stromal Vascular Fraction from Murine White Adipose Tissue Using a Tissue Dissociator (Video) | JoVE 

The in vitro study of white, brown, and beige adipocyte differentiation enables the investigation of cell-autonomous functions of adipocytes and their ...