Name: fabrication and implantation of miniature dual-element strain gages for measuring in vivo gastrointestinal contractions in rodents.
Uploaded: 2014-09-18T14:00:00
Description: Watch this Scientific Journal Video about Fabrication and Implantation of Miniature Dual-element Strain Gages for Measuring In Vivo Gastrointestinal Contractions in Rodents. at JoVE.com

Research funding was received through the National Institute of Neurological Disorders and Stroke &#160;(NS049177 and NS087834). The authors wish&#160;to acknowledge the intellectual contributions of the late Dr. Paul Bass and his colleagues to the original design of the strain gages; and Carol Tollefsrud for the fabrication and marketing of the strain gages until the cessation of production in 2010 as well as for her insightful correspondence.

All procedures followed National Institutes of Health guidelines and were approved by the Institutional Animal Care and Use Committee at the Penn State Hershey College of Medicine. Rats were housed using common vivarium practices. Note: This protocol uses male Wistar rats &#8805;8 weeks of age and initially weighing 175 - 200 g.
1. Procedures for Fabrication of Strain Gage
<ol>
	<li>Most tooling and components remain available from the original or successor companies and are summarized in <s...

<ol>
	<li>Szarka, L. A., Camilleri, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+measurement+of+gastric+motility.">Methods for measurement of gastric motility.</a> Am. J. Physiol. Gastrointest. Liver Physiol. 296 (3), G461-G475 (2009).</li><li>Pascaud XB, F. A. U., Genton, M. J., Bass, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+miniature+transducer+for+recording+intestinal+motility+in+unrestrained+chronic+rats.">A miniature transducer for recording intestinal motility in unrestrained chronic rats.</a> Am. J. Physiol. Endocrinol. Metab. Gastrointest. Physiol. 4 (5), 532-538 (1978).</li><li>Gourcerol, G., Adelson, D. W., Million, M., Wang, L., Tache, Y. <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+gastric+motility+by+brain-gut+peptides+using+a+novel+non-invasive+miniaturized+pressure+transducer+method+in+anesthetized+rodents.">Modulation of gastric motility by brain-gut peptides using a novel non-invasive miniaturized pressure transducer method in anesthetized rodents.</a> Peptides. 32 (4), 737-746 (2011).</li><li>Am&#233;rico, M. F., 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=Validation+of+ACB+in+vitro+and+in+vivo+as+a+biomagnetic+method+for+measuring+stomach+contraction.">Validation of ACB in vitro and in vivo as a biomagnetic method for measuring stomach contraction.</a> Neurogastroenterol. Motil. 22 (12), 1340-1374 (2010).</li><li>Fujitsuka, N., Asakawa, A., Amitani, H., Fujimiya, M., Inui, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+Eighteen+-+Ghrelin+and+Gastrointestinal+Movement.">Chapter Eighteen - Ghrelin and Gastrointestinal Movement.</a> Ghrelin and Gastrointestinal Movement. , 289-301 (2012).</li><li>Monroe, M. J., Hornby, P. J., Partosoedarso, E. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Central+vagal+stimulation+evokes+gastric+volume+changes+in+mice:+a+novel+technique+using+a+miniaturized+barostat.">Central vagal stimulation evokes gastric volume changes in mice: a novel technique using a miniaturized barostat.</a> Neurogastroenterol. Motil. 16 (1), 5-11 (2004).</li><li>Herman, M. 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=Characterization+of+noradrenergic+transmission+at+the+dorsal+motor+nucleus+of+the+vagus+involved+in+reflex+control+of+fundus+tone.">Characterization of noradrenergic transmission at the dorsal motor nucleus of the vagus involved in reflex control of fundus tone.</a> Am. J. Physiol. Regul. Integr. Comp. Physiol. 294 (3), 720-729 (2008).</li><li>Gondim, F. 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=Complete+cervical+or+thoracic+spinal+cord+transections+delay+gastric+emptying+and+gastrointestinal+transit+of+liquid+in+awake+rats.">Complete cervical or thoracic spinal cord transections delay gastric emptying and gastrointestinal transit of liquid in awake rats.</a> Spinal Cord. 37 (11), 793-799 (1999).</li><li>Van Bree, S. H. W., 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=Systemic+inflammation+with+enhanced+brain+activation+contributes+to+more+severe+delay+in+postoperative+ileus.">Systemic inflammation with enhanced brain activation contributes to more severe delay in postoperative ileus.</a> Neurogastroenterol. Motil. 25 (8), 540-549 (2013).</li><li>Qualls-Creekmore, E., Tong, M., Holmes, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastric+emptying+of+enterally+administered+liquid+meal+in+conscious+rats+and+during+sustained+anaesthesia.">Gastric emptying of enterally administered liquid meal in conscious rats and during sustained anaesthesia.</a> Neurogastroenterol. Motil. 22 (2), 181-185 (2010).</li><li>Qualls-Creekmore, E., Tong, M., Holmes, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time-course+of+recovery+of+gastric+emptying+and+motility+in+rats+with+experimental+spinal+cord+injury.">Time-course of recovery of gastric emptying and motility in rats with experimental spinal cord injury.</a> Neurogastroenterol. Motil. 22 (1), 62 (2010).</li><li>Choi, K. 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=Determination+of+gastric+emptying+in+nonobese+diabetic+mice.">Determination of gastric emptying in nonobese diabetic mice.</a> Am. J. Physiol. Gastrointest. Liver Physiol. 293 (5), G1039-G1045 (2007).</li><li>Adelson, D. W., Million, M., Kanamoto, K., Palanca, T., Tache, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coordinated+gastric+and+sphincter+motility+evoked+by+intravenous+CCK-8+as+monitored+by+ultrasonomicrometry+in+rats.">Coordinated gastric and sphincter motility evoked by intravenous CCK-8 as monitored by ultrasonomicrometry in rats.</a> Am. J. Physiol. Gastrointest. Liver Physiol. 286 (2), G321-G332 (2004).</li><li>Xue, 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=Effect+of+modulation+of+serotonergic,+cholinergic,+and+nitrergic+pathways+on+murine+fundic+size+and+compliance+measured+by+ultrasonomicrometry.">Effect of modulation of serotonergic, cholinergic, and nitrergic pathways on murine fundic size and compliance measured by ultrasonomicrometry.</a> Am. J. Physiol. Gastrointest. Liver Physiol. 290 (1), G74-G82 (2005).</li><li>Bass, P., Wiley, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contractile+force+transducer+for+recording+muscle+activity+in+unanesthetized+animals.">Contractile force transducer for recording muscle activity in unanesthetized animals.</a> J. Appl. Physiol. 32 (4), 567-570 (1972).</li><li>Holmes, G. M., Browning, K. N., Tong, M., Qualls-Creekmore, E., Travagli, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vagally+mediated+effects+of+glucagon-like+peptide+1:+in+vitro+and+in+vivo+gastric+actions.">Vagally mediated effects of glucagon-like peptide 1: in vitro and in vivo gastric actions.</a> J. Physiol. 587 (19), 4749-4759 (2009).</li><li>Tong, M., Qualls-Creekmore, E., Browning, K. N., Travagli, R. A., Holmes, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+spinal+cord+injury+in+rats+diminishes+vagally-mediated+gastric+responses+to+cholecystokinin-8s.">Experimental spinal cord injury in rats diminishes vagally-mediated gastric responses to cholecystokinin-8s.</a> Neurogastroenterol. Motil. 23 (2), e69-e79 (2011).</li><li>Miyano, 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=The+role+of+the+vagus+nerve+in+the+migrating+motor+complex+and+ghrelin-+and+motilin-induced+gastric+contraction+in+suncus.">The role of the vagus nerve in the migrating motor complex and ghrelin- and motilin-induced gastric contraction in suncus.</a> PLoS ONE. 8 (5), e64777 (2013).</li><li>Holmes, G. M., Rogers, R. C., Bresnahan, J. C., Beattie, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thyrotropin-releasing+hormone+(TRH)+and+CNS+regulation+of+anorectal+motility+in+the+rat.">Thyrotropin-releasing hormone (TRH) and CNS regulation of anorectal motility in the rat.</a> J Auton. Nerv. Syst. 56, 8-14 (1995).</li><li>Ormsbee, H. S., Bass, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastroduodenal+motor+gradients+in+the+dog+after+pyloroplasty.">Gastroduodenal motor gradients in the dog after pyloroplasty.</a> Am. J. Physiol. 230, 389-397 (1976).</li><li>Fukuda, H., 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=Impaired+gastric+motor+activity+after+abdominal+surgery+in+rats.">Impaired gastric motor activity after abdominal surgery in rats.</a> Neurogastroenterol. Motil. 17 (2), 245-250 (2005).</li><li>Browning, K. N., Babic, T., Holmes, G. M., Swartz, E., Travagli, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+critical+re-evaluation+of+the+specificity+of+action+of+perivagal+capsaicin.">A critical re-evaluation of the specificity of action of perivagal capsaicin.</a> J. Physiol. 591 (6), 1563-1580 (2013).</li></ol>

The procedures presented here allow individual laboratories to fabricate sensitive miniature strain gages for biological applications including, but not limited to, gastrointestinal motility in small laboratory animals. Since the commercial manufacture of these strain gages has ceased, laboratories investigating gastrointestinal function are limited to other techniques which may not permit the full range of experimental applications that are available. This report provides an updated and more detailed description of prev...

Experimental studies that record in vivo gastrointestinal (GI) motility across a number of experimental conditions remain a powerful tool for understanding the underlying normal and pathophysiological processes necessary for nutrient homeostasis. Traditionally, numerous experimental methodologies, some with similarities to those found in clinical practice 1, have been employed to directly quantify changes in GI contraction rate 2-5, intraluminal pressure 6, 7, or the GI transit of non-absorbable markers 8, 9 or stable isotopes 10-12. Each of these techniques has unique advantages and disadvantages, which have been addressed previously in the literature. For example, the utility of balloon manometry to quantify pressure changes has been questioned due to the inherent compliance of the balloon material while gastrointestinal recovery of nonabsorbable markers requires euthanizing the experimental animal for a single data point. Recently, the application and validation of a miniaturized arterial pressure catheter has been reported that offers a non-surgical method for monitoring gastric contractility in rats and mice 3. While an orogastrically placed pressure transducer effectively eliminates confounding variables on gastrointestinal function by avoiding invasive surgical procedures, such an approach is only suitable for anesthetized preparations. Furthermore, the lack of visual guidance does not permit consistent placement of the transducer within specific regions of the stomach. As such, this application is restricted to the stomach or colon since visualization, coupled with the relatively stiff transducer wire, within the duodenum or ileum is not an option.
Similarly, the biomagnetic alternate current biosusceptometry (ACB) technique has been validated for GI contraction analysis 4. While the ACB technique provides a noninvasive approach for measuring gastrointestinal contractions, ACB suffers from a similar limitation in that the use of ingested magnetic detection media does not permit precise recording of specific regions of the GI tract. This limitation can be overcome through the surgical implantation of magnetic markers. Nonetheless, the ACB technique necessitates that the animal be anesthetized for data collection.
Ultrasonomicrometry has been employed in some GI studies 13, 14 in order to take advantage of the small size, spatial, and temporal advantages of piezoelectric crystal transmitter/receivers. Waves of gastric smooth muscle contraction are not a high-frequency event and occur at a rate of approximately 3 - 5 cycles/min. Therefore, the temporal advantages of sonomicrometry may be unnecessary to justify the cost. Furthermore, while linear motion is accurately measured with sonomicrometry, limitations have been presented regarding accurate gastrointestinal data interpretation that may result from implanting an insufficient number of crystals 14.
Based upon the original designs of Bass and colleagues 2, 15 this visualized protocol more fully documents the step-by-step fabrication and experimental application of miniature, dual element strain gages that possess high sensitivity and flexibility for recording smooth muscle contractions along the entire GI tract. The dimensions of the strain gage elements are suitable for any rodent application since sensitivity and size of the finished strain gage are most dependent upon the silicone sheets encapsulating the elements. These strain gages are readily adapted for acute and chronic application in anesthetized and freely behaving laboratory animal models thereby providing a single technique for quantifying smooth muscle contractions.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Strain gage element</td>
			<td>Micro-Measurements (Vishay Product Group)</td>
			<td>EA-06-031-350</td>
			<td>&#160;Linear pattern, foil, stress analysis strain gage (2 required) 
			<a href="http://www.vishaypg.com/micro-measurements/">www.vishaypg.com/micro-measurements/</a> 
			or 
			<a href="http://www.vishaypg.com/docs/11070/031ce.pdf">http://www.vishaypg.com/docs/11070/031ce.pdf</a></td>
		</tr>
		<tr>
			<td>epoxy-phenolic adhesive</td>
			<td>M-bond 610</td>
			<td>General purpose adhesive for bonding strain gage elements</td>
			<td><a href="http://www.vishaypg.com/docs/11024/wirecable.pdf">http://www.vishaypg.com/docs/11024/wirecable.pdf</a></td>
		</tr>
		<tr>
			<td>3 conductor insulated wire</td>
			<td>336-FTE</td>
			<td>Fine gage, flexible general purpose wire</td>
			<td><a href="http://www.vishaypg.com/docs/11024/wirecable.pdf">http://www.vishaypg.com/docs/11024/wirecable.pdf</a></td>
		</tr>
		<tr>
			<td>Flux and rosin solvent kit</td>
			<td>FAR-2 M-Flux AR kit</td>
			<td>Liquid solder flux</td>
			<td><a href="http://www.vishaypg.com/docs/11023/soldacce.pdf">http://www.vishaypg.com/docs/11023/soldacce.pdf</a></td>
		</tr>
		<tr>
			<td>Solder</td>
			<td>361A-20R-25</td>
			<td>Optimized and recommended for strain gage applications</td>
			<td><a href="http://www.vishaypg.com/docs/11023/soldacce.pdf">http://www.vishaypg.com/docs/11023/soldacce.pdf</a></td>
		</tr>
		<tr>
			<td>Gold socket connector</td>
			<td>PlasticsOne</td>
			<td>E363/0</td>
			<td>Socket contact for electrode pedestal 
			<a href="http://www.plastics1.com/PCR/Catalog/Item.php?item=407">http://www.plastics1.com/PCR/Catalog/Item.php?item=407</a></td>
		</tr>
		<tr>
			<td>Electrode pedestal</td>
			<td>MS363</td>
			<td>Secure platform for wire contacts</td>
			<td><a href="http://www.plastics1.com/PCR/Catalog/Item.php?item=499">http://www.plastics1.com/PCR/Catalog/Item.php?item=499</a></td>
		</tr>
		<tr>
			<td>6-wire cable</td>
			<td>363 PLUG W/VINYL SL/6</td>
			<td>Pre-fabricated vinyl-coated cable (in customized lengths) with plug adaptor to match electrode pedestal and tinned solder lugs on terminal end</td>
			<td></td>
		</tr>
		<tr>
			<td>Silicone rubber casting compound</td>
			<td>EIS electrical products</td>
			<td>Elan Tron E211</td>
			<td>Potting medium for gage/wire solder joints 
			<a href="http://www.eis-inc.com/">http://www.eis-inc.com</a></td>
		</tr>
		<tr>
			<td>HOTweezers</td>
			<td>Meisei Corporation</td>
			<td>Model 4B</td>
			<td>Wire insulation strippers 
			<a href="http://www.impexron.us/">http://www.impexron.us</a></td>
		</tr>
		<tr>
			<td>Soldering station</td>
			<td>Weller (Apex Tool Group)</td>
			<td>WES 51</td>
			<td>High quality soldering equipment 
			<a href="http://www.apexhandtools.com/weller/index.cfm">http://www.apexhandtools.com/weller/index.cfm</a></td>
		</tr>
		<tr>
			<td>Available through http://www.eis-inc.com or http://www.amazon.com</td>
		</tr>
		<tr>
			<td>Silicone sheet</td>
			<td>Trelleborg Sealing Solutions Northborough-Life Sciences</td>
			<td>Pharmelast 20-20</td>
			<td>Encapsulating strain gauge elements 
			10 B Forbes Road Northborough, MA 01532 (800) 634-2000</td>
		</tr>
		<tr>
			<td>Amplifier</td>
			<td>Experimetria Ltd</td>
			<td>AMP-01-SG</td>
			<td> 
			<a href="http://experimetria.com/Biological_amplifiers.php">http://experimetria.com/Biological_amplifiers.php</a></td>
		</tr>
	</tbody>
</table>

fabrication and implantation of miniature dual-element strain gages for measuring in vivo gastrointestinal contractions in rodents.

Gastrointestinal dysfunction remains a major cause of morbidity and mortality. Indeed, gastrointestinal (GI) motility in health and disease remains an area of productive research with over 1,400 published animal studies in just the last 5 years. Numerous techniques have been developed for quantifying smooth muscle activity of the stomach, small intestine, and colon. In vitro and ex vivo techniques offer powerful tools for mechanistic studies of GI function, but outside the context of the integrated systems inherent to an intact organism. Typically, measuring in vivo smooth muscle contractions of the stomach has involved an anesthetized preparation coupled with the introduction of a surgically placed pressure sensor, a static pressure load such as a mildly inflated balloon or by distending the stomach with fluid under barostatically-controlled feedback. Yet many of these approaches present unique disadvantages regarding both the interpretation of results as well as applicability for in vivo use in conscious experimental animal models. The use of dual element strain gages that have been affixed to the serosal surface of the GI tract has offered numerous experimental advantages, which may continue to outweigh the disadvantages. Since these gages are not commercially available, this video presentation provides a detailed, step-by-step guide to the fabrication of the current design of these gages. The strain gage described in this protocol is a design for recording gastric motility in rats. This design has been modified for recording smooth muscle activity along the entire GI tract and requires only subtle variation in the overall fabrication. Representative data from the entire GI tract are included as well as discussion of analysis methods, data interpretation and presentation.

Representative data from a Thiobutabarbital-anesthetized rat are shown in Figure 2. The top trace represents the gastric corpus contractions from the rat during the brainstem administration of thyrotropin releasing hormone (TRH, 100 pmol), a known motility-enhancing peptide 3, 19. It shows baseline contractions prior to the increase in phasic gastric smooth muscle activity. Note: Analysis of these peaks in gastric contractions follow the original formula devised by Ormsby and Bass 20</sup...

Watch this Scientific Journal Video about Fabrication and Implantation of Miniature Dual-element Strain Gages for Measuring In Vivo Gastrointestinal Contractions in Rodents. at JoVE.com

Fabrication and Implantation of Miniature Dual-element Strain Gages for Measuring In Vivo Gastrointestinal Contractions in Rodents.

The in vivo measurement of smooth muscle contractions along the gastrointestinal tract of laboratory animals remains a powerful, though underutilized, technique. Flexible, dual element strain gages are not commercially available and require fabrication. This protocol describes the construction of reliable, inexpensive strain gages for acute or chronic implantation in rodents.

Fabrication and Implantation of Miniature Dual-element Strain Gages for Measuring In Vivo Gastrointestinal Contractions in Rodents.

Gastrointestinal dysfunction remains a major cause of morbidity and mortality. Indeed, gastrointestinal (GI) motility in health and disease remains an ...

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