Name: mesenteric artery contraction and relaxation studies using automated wire myography
Uploaded: 2011-09-22T13:00:00
Description: Watch this Scientific Journal Video about Mesenteric Artery Contraction and Relaxation Studies Using Automated Wire Myography at JoVE.com

The project described was supported by Award Numbers R01 HL064761, R25 HL059868, 1SC1 HL099139 and P20 MD000175 form National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

1. Isolation of rat mesenteric small artery
<ol>
 <li>Anesthetize animal with isoflurane in a closed chamber and wipe the abdomen with alcohol.</li>
 <li>Perform a mid-line laparotomy to expose mesenteric bed.</li>
 <li>Using scissors, remove about 85 cm of intestine with feeding vasculature with the superior mesenteric artery. Cut the proximal end of intestinal section close to the pylorus and the distal end near the ileo-coecal junction. Segments of the intestine are isolated from rats that are deeply anesthetized wi...

<ol>
	<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, 617-619 (1976).</li><li>Bevan, J. A., Osher, 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=A+direct+method+for+recording+tension+changes+in+the+wall+of+small+blood+vessels+in+vitro.">A direct method for recording tension changes in the wall of small blood vessels in vitro.</a> Agents Actions. 2, 257-260 (1972).</li><li>Mulvany, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Procedures+for+investigating+of+small+vessels+using+small+vessel+myograph.">Procedures for investigating of small vessels using small vessel myograph.</a> DMT Danish Myo Technology. , (2004).</li><li>Angus, J. A., Wright, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Techniques+to+study+the+pharmacodynamics+of+isolated+large+and+small+blood+vessels.">Techniques to study the pharmacodynamics of isolated large and small blood vessels.</a> J. Pharmacol. Toxicol. Methods. 44, 395-407 (2000).</li><li>Halpern, W., Mulvany, M. J., Warshaw, D. M. <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+smooth+muscle+cells+in+the+walls+of+arterial+resistance+vessels.">Mechanical properties of smooth muscle cells in the walls of arterial resistance vessels.</a> J. Physiol. 275, 85-101 (1978).</li><li>Lindhorst, J., Alexander, N., Blignaut, J., Rayner, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differences+in+hypertension+between+blacks+and+whites:+an+overview.">Differences in hypertension between blacks and whites: an overview.</a> Cardiovasc. J. Afr. 18, 241-247 (2007).</li><li>Eley, S. L., Allen, C. M., Williams, C. L., Bukosi, R. D., Pointer, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Action+of+thiazide+on+renal+interstitial+calcium.">Action of thiazide on renal interstitial calcium.</a> Am. J. Hypertens. 21, 814-819 (2008).</li><li>Palmer, C. E., Rudd, M. A., Bujoski, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Renal+interstitial+Ca2++during+sodium+loading+of+normotensive+and+Dal-salt+hypertensive+rats.">Renal interstitial Ca2+ during sodium loading of normotensive and Dal-salt hypertensive rats.</a> Am. J. Hypertens. 16, 771-776 (2003).</li><li>Hurwitz, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Homeostatic+control+of+plasma+calcium+concentration.">Homeostatic control of plasma calcium concentration.</a> Crit. Rev. Biochem. Mol. Biol. 31, 41-100 (1996).</li><li>Brown, E. M., MacLeod, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+sensing+and+extracellular+calcium+signaling.">Extracellular sensing and extracellular calcium signaling.</a> Physiol. Rev. 81, 239-297 (2001).</li><li>Breitwieser, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+calcium+as+an+integrator+of+tissue+function.">Extracellular calcium as an integrator of tissue function.</a> Int. J. Biochem. Cell Biol. 40, 1467-1480 (2008).</li><li>Mupanomunda, M. M., Wang, Y., Bukoski, R. D. <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+chronic+sensory+denervation+on+Ca2+-induced+relaxation+of+isolated+mesenteric+resistance+arteries.">Effect of chronic sensory denervation on Ca2+-induced relaxation of isolated mesenteric resistance arteries.</a> Am. J. Physiol. 274, 1655-1661 (1998).</li><li>Mupanomunda, M. M., Ishioka, N., Bukoski, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interstitial+Ca2++undergoes+dynamic+changes+sufficient+to+stimulate+nerve-dependent+Ca2+-induced+relaxation.">Interstitial Ca2+ undergoes dynamic changes sufficient to stimulate nerve-dependent Ca2+-induced relaxation.</a> Am. J. Physiol. 276, 1035-1042 (1999).</li><li>Mupanomunda, M. M., Tian, B., Ishioka, N., Bukoski, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Renal+interstitial+Ca2+.">Renal interstitial Ca2+.</a> Am. J. Physiol. Renal Physiol. 278, F644-F649 (2000).</li><li>Awumey, E. M., Hill, S. K., Diz, D. I., Bukoski, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytochrome+P-450+metabolites+of+2-arachidonoylglycerol+play+a+role+in+Ca2+-induced+relaxation+of+rat+mesenteric+arteries.">Cytochrome P-450 metabolites of 2-arachidonoylglycerol play a role in Ca2+-induced relaxation of rat mesenteric arteries.</a> Am. J. Physiol. Heart Circ. Physiol. 294, 2363-2370 (2008).</li><li>Chen, W., Bergsman, J. B., Wang, X., Gilkey, G., Pierpoint, C. R., Daniel, E. A., Awumey, E. M., Dauban, P., Dodd, R. H., Ruat, M., Smith, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Presynaptic+external+calcium+signaling+involves+the+calcium-sensing+receptor+in+neocortical+nerve+terminals.">Presynaptic external calcium signaling involves the calcium-sensing receptor in neocortical nerve terminals.</a> PloS One. 5, e8563-e8563 (2010).</li><li>Bukoski, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+perivascular+sensory+nerve+Ca2++receptor+and+blood+pressure+regulation:+a+hypothesis.">The perivascular sensory nerve Ca2+ receptor and blood pressure regulation: a hypothesis.</a> Am. J. Hypertens. 11, 1117-1123 (1998).</li><li>Bukoski, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dietary+Ca2++and+blood+pressure:+evidence+that+Ca2+-sensing+receptor+activated+sensory+nerve+dilator+activity+couples+changes+in+interstitial+Ca2++with+vascular.">Dietary Ca2+ and blood pressure: evidence that Ca2+-sensing receptor activated sensory nerve dilator activity couples changes in interstitial Ca2+ with vascular.</a> 16, 218-221 (2001).</li></ol>

Hypertension is a leading cause of cardiovascular, cerebral and renal morbidity/mortality. The occurrence of hypertension is high in the population and salt-sensitive hypertension is particularly high in the aging population, and more prevalent in blacks than whites. This is believed to be due to the tendency of blacks to retain sodium in their kidneys9. Salt-sensitivity is a major contributing factor to kidney disease and is associated with endothelial dysfunction, but the mechanism is not fully understood. R...

<table border="1">
	<tbody>
		<tr>
			<th>Name of equipment/ reagent	</th>
			<th>Company	</th>
			<th>Catalogue number	</th>
			<th>Comments(optional)</th>
		</tr>
		<tr>
			<td>Auto Dual Wire Myograph System-510A	DMT-USA, Inc.</td>
			<td>Atlanta, GA.</td>
			<td>100151</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td><a href="http://www.adinstruments.com/products/hardware/corporate/product/ML866/" target="_blank">PowerLab/4SP Data Acquisition System</a></td>
			<td><a href="http://www.adinstruments.com/" target="_blank">ADInstruments</a>, Colorado Springs, CO.</td>
			<td><a href="http://www.adinstruments.com/products/hardware/corporate/product/ML866/" target="_blank">ML750</a></td>
			<td>New models with 4-16 input channels are available.</td>
		</tr>
		<tr>
			<td>Dell Dimension XPS Gen 4 Computer	</td>
			<td>Dell</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Stemi SV II (Apo) Dissection Microscope with Ocular	</td>
			<td>Carl Zeiss International.</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Wistar, Dahl salt-sensitive, Dahl salt-resistant rats	</td>
			<td>Harlan Sprague Dawley, Indianapolis, IN.</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Rodent chow	</td>
			<td>Harland Tekland, Madison, WI.</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Phenylephrine</td>
			<td>Sigma-Aldrich, St. Louis, MO.</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Other chemicals</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>All chemicals used were of the purest grades available commercially.</td>
		</tr>
	</tbody>
</table>

mesenteric artery contraction and relaxation studies using automated wire myography

Proximal resistance vessels, such as the mesenteric arteries, contribute substantially to the peripheral resistance. These small vessels of between 100-400 &mu;m in diameter function primarily in directing blood flow to various organs according to the overall requirements of the body. The rat mesenteric artery has a diameter greater than 100 &mu;m. The myography technique, first described by Mulvay and Halpern1, was based on the method proposed by Bevan and Osher2. The technique provides information about small vessels under isometric conditions, where substantial shortening of the muscle preparation is prevented. Since force production and sensitivity of vessels to different agonists is dependent on the extent of stretch, according to active tension-length relation, it is essential to conduct contraction studies under isometric conditions to prevent compliance of the mounting wires. Stainless steel wires are preferred to tungsten wires because of oxidation of the latter, which affects recorded responses3.The technique allows for the comparison of agonist-induced contractions of mounted vessels to obtain evidence for normal function of vascular smooth muscle cell receptors.
We have shown in several studies that isolated mesenteric arteries that are contracted with phenylyephrine relax upon addition of cumulative concentrations of extracellular calcium (Ca2+e). The findings led us to conclude that perivascular sensory nerves, which express the G protein-coupled Ca2+-sensing receptor (CaR), mediate this vasorelaxation response. Using an automated wire myography method, we show here that mesenteric arteries from Wistar, Dahl salt-sensitive(DS) and Dahl salt-resistant (DR) rats respond differently to Ca2+e. Tissues from Wistar rats showed higher Ca2+-sensitivity compared to those from DR and DS. Reduced CaR expression in mesenteric arteries from DS rats correlates with reduced Ca2+e-induced relaxation of isolated, pre-contracted arteries. The data suggest that the CaR is required for relaxation of mesenteric arteries under increased adrenergic tone, as occurs in hypertension, and indicate an inherent defect in the CaR signaling pathway in Dahl animals, which is much more severe in DS.
The method is useful in determining vascular reactivity ex vivo in mesenteric resistance arteries and similar small blood vessels and comparisons between different agonists and/or antagonists can be easily and consistently assessed side-by-side6,7,8.

Watch this Scientific Journal Video about Mesenteric Artery Contraction and Relaxation Studies Using Automated Wire Myography at JoVE.com

Mesenteric Artery Contraction and Relaxation Studies Using Automated Wire Myography

An automated myography method for force measurements in isolated mesenteric arteries is described. It employs a Mulvany-Halpern Auto Dual Wire Myograph 510A to determine responses to phenylephrine and extracellular calcium. The method allows consistent determination of isometric responses to agonists in small vessels of diameters of 60 - 300 &mu;m, independently.

Proximal resistance vessels, such as the mesenteric arteries, contribute substantially to the peripheral resistance. These small vessels of between ...

Research

JoVE Journal

Medicine

Guide Wire Assisted Catheterization and Colored Dye Injection for Vascular Mapping of Monochorionic Twin Placentas

Monochorionic (MC) twin pregnancies are associated with significantly higher morbidity and mortality rates than dichorionic twins. Approximately 50% of MC ...

Assessing Myogenic Response and Vasoactivity In Resistance Mesenteric Arteries Using Pressure Myography

Small resistance arteries constrict and dilate respectively in response to increased or decreased intraluminal pressure; this phenomenon known as myogenic ...

Isolating and Using Sections of Bovine Mesenteric Artery and Vein as a Bioassay to Test for Vasoactivity in the Small Intestine

Mammalian gastrointestinal systems are constantly exposed to compounds (desirable and undesirable) that can have an effect on blood flow to and from that ...

Murine Model of Femoral Artery Wire Injury with Implantation of a Perivascular Drug Delivery Patch

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. ...

A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

Laparoscopic Technique for Serial Collection of Liver and Mesenteric Lymph Nodes in Macaques

The mesenteric lymph nodes (MLN) and the liver are exposed to microbes and microbial products from the gastrointestinal (GI) tract, making them ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Brachial Artery Catheterization in Swine

The video describes in detail the catheterization of the distal brachial artery in swine. This technique enables researchers to measure arterial blood ...

Murine Myocardial Infarction Model using Permanent Ligation of Left Anterior Descending Coronary Artery

Myocardial infarction (MI) and acute coronary diseases are among the most prominent causes of death in population with western lifestyle. The murine ...