Name: rapid determination of the thermal nociceptive threshold in diabetic rats
Uploaded: 2012-05-17T13:00:00
Description: Watch this Scientific Journal Video about Rapid Determination of the Thermal Nociceptive Threshold in Diabetic Rats at JoVE.com

This work was funded by the American Diabetes Association (ADA), Grant JF1-10-14 (MDiF). We would like to thank the personnel of the Laboratory of Animal Resources at WSU. Authors gratefully acknowledge assistance with the statistical analysis of data from Neil Paton, Ph.D.

TNTs of mice and rats are determined by using the incremental hot-plate analgesia meter [iHPAM, IITC Inc. Life Science (Woodland Hills, CA)]. The equipment consists of several components: an aluminum plate (10 x 20 cm) with the heating system underneath and a Plexiglas observation chamber above; a heat controlling unit, software for data acquisition (IITC Part # Soft Series 8), a PC (personal computer) and a footswitch for remote start, stop or reset control of the unit. The heating system also allows starting/stopping/r...

<ol>
	<li>Baron, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peripheral+neuropathic+pain:+From+mechanisms+to+symptoms.">Peripheral neuropathic pain: From mechanisms to symptoms.</a> Clin. J. Pain. 16, S12-S20 (2000).</li><li>Calcutt, N. A., Jorge, M. C., Yaksh, T. L., Chaplan, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tactile+allodynia+and+formalin+hyperalgesia+in+streptozotocin-diabetic+rats:+Effects+of+insulin,+aldose+reductase+inhibition+and+lidocaine.">Tactile allodynia and formalin hyperalgesia in streptozotocin-diabetic rats: Effects of insulin, aldose reductase inhibition and lidocaine.</a> Pain. 68, 293-299 (1996).</li><li>Bolcskei, K., Petho, G., Szolcsanyi, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noxious+heat+threshold+measured+with+slowly+increasing+temperatures:+Novel+rat+thermal+hyperalgesia+models.">Noxious heat threshold measured with slowly increasing temperatures: Novel rat thermal hyperalgesia models.</a> Methods Mol. Biol. 617, 57-66 (2010).</li><li>Ankier, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+hot+plate+tests+to+quantify+antinociceptive+and+narcotic+antagonist+activities.">New hot plate tests to quantify antinociceptive and narcotic antagonist activities.</a> Eur. J. Pharmacol. 27, 1-4 (1974).</li><li>Almasi, R., Petho, G., Bolcskei, K., Szolcsanyi, J. <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+resiniferatoxin+on+the+noxious+heat+threshold+temperature+in+the+rat:+A+novel+heat+allodynia+model+sensitive+to+analgesics.">Effect of resiniferatoxin on the noxious heat threshold temperature in the rat: A novel heat allodynia model sensitive to analgesics.</a> Br. J. Pharmacol. 139, 49-58 (2003).</li><li>Bolcskei, K., Horvath, D., Szolcsanyi, J., Petho, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heat+injury-induced+drop+of+the+noxious+heat+threshold+measured+with+an+increasing-temperature+water+bath:+A+novel+rat+thermal+hyperalgesia+model.">Heat injury-induced drop of the noxious heat threshold measured with an increasing-temperature water bath: A novel rat thermal hyperalgesia model.</a> Eur. J. Pharmacol. 564, 80-87 (2007).</li><li>Chesler, E. J., Wilson, S. G., Lariviere, W. R., Rodriguez-Zas, S. L., Mogil, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+and+ranking+of+genetic+and+laboratory+environment+factors+influencing+a+behavioral+trait,+thermal+nociception,+via+computational+analysis+of+a+large+data+archive.">Identification and ranking of genetic and laboratory environment factors influencing a behavioral trait, thermal nociception, via computational analysis of a large data archive.</a> Neurosci. Biobehav. Rev. 26, 907-923 (2002).</li><li>Langford, D. J., Crager, S. E., Shehzad, Z., Smith, S. B., Sotocinal, S. G., Levenstadt, J. S., Chanda, M. L., Levitin, D. J., Mogil, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+modulation+of+pain+as+evidence+for+empathy+in+mice.">Social modulation of pain as evidence for empathy in mice.</a> Science. 312, 1967-1970 (2006).</li><li>Hunt, S. P., Koltzenburg, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The neurobiology of pain. , (2005).</li><li>Willis, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The pain system : The neural basis of nociceptive transmission in the mammalian nervous system. , (1985).</li><li>Calcutt, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Modeling diabetic sensory neuropathy in rats. In: Methods in molecular medicine. Pain research: Methods and protocols. , (2004).</li><li>Bars, D. L. e., Gozariu, M., Cadden, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+nociception.">Animal models of nociception.</a> Pharmacol. Rev. 53, 597-652 (2001).</li><li>Shaikh, A. S., Somani, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+and+biomarkers+of+neuropathy+in+diabetic+rodents.">Animal models and biomarkers of neuropathy in diabetic rodents.</a> Indian J. Pharmacol. 42, 129-134 (2010).</li><li>Hargreaves, K., Dubner, R., Brown, F., Flores, C., Joris, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+and+sensitive+method+for+measuring+thermal+nociception+in+cutaneous+hyperalgesia.">A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia.</a> Pain. 32, 77-88 (1988).</li><li>Hardy, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Method+for+the+rapid+measurement+of+skin+temperature+during+exposure+to+intense+thermal+radiation.">Method for the rapid measurement of skin temperature during exposure to intense thermal radiation.</a> J. Appl. Physiol. 5, 559-566 (1953).</li><li>Sumino, R., Dubner, R., Starkman, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Responses+of+small+myelinated+&amp;quot;warm&amp;quot;+fibers+to+noxious+heat+stimuli+applied+to+the+monkey's+face.">Responses of small myelinated &amp;quot;warm&amp;quot; fibers to noxious heat stimuli applied to the monkey's face.</a> Brain Res. 62, 260-263 (1973).</li><li>Hammond, D. L., Ruda, 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=Developmental+alterations+in+thermal+nociceptive+threshold+and+the+distribution+of+immunoreactive+calcitonin+gene-related+peptide+and+substance+p+after+neonatal+administration+of+capsaicin+in+the+rat.">Developmental alterations in thermal nociceptive threshold and the distribution of immunoreactive calcitonin gene-related peptide and substance p after neonatal administration of capsaicin in the rat.</a> Neurosci. Lett. 97, 57-62 (1989).</li></ol>

Similarly to the classic hot plate test to quantify thermal hyperalgesia4,13, the assay of nociception described here permits a fast and reliable way to quantify nocifensive behavior in rats and mice. However, contrary to classic test, the incremental hot-plate method is non-invasive and virtually stress-free. Although some restrain is necessary to perform the test (i.e., the animal must be in the observation chamber), rats or mice are accustomed to similar areas (e.g., housing cages).
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<table border="1">
	<tbody>
 <tr>
 <td>Name</td>
 <td>Company</td>
 <td>Catalogue number</td>
 </tr>
 <tr>
 <td>Incremental Hot-Plate Analgesia Meter </td>
 <td>IITC Inc. Life Science </td>
 <td>Part #PE34 </td>
 </tr>
 <tr>
 <td>Soft Series 8</td>
 <td>IITC Inc. Life Science</td>
 <td>Part # Series8</td>
 </tr>
 <tr>
 <td>Streptozotocin</td>
 <td>Calbiochem</td>
 <td>572201</td>
 </tr>
 </tbody>
 </table>

rapid determination of the thermal nociceptive threshold in diabetic rats

Painful diabetic neuropathy (PDN) is characterized by hyperalgesia i.e., increased sensitivity to noxious stimulus, and allodynia i.e., hypersensitivity to normally innocuous stimuli1. Hyperalgesia and allodynia have been studied in many different rodent models of diabetes mellitus2. However, as stated by B&ouml;lcskei et al, determination of "pain" in animal models is challenging due to its subjective nature3. Moreover, the traditional methods used to determine behavioral responses to noxious thermal stimuli usually lack reproducibility and pharmacological sensitivity3. For instance, by using the hot-plate method of Ankier4, flinch, withdrawal and/or licking of either hind- and/or fore-paws is quantified as reflex latencies at constant high thermal stimuli (52-55 &deg;C). However, animals that are hyperalgesic to thermal stimulus do not reproducibly show differences in reflex latencies using those supra-threshold temperatures3,5. As the recently described method of B&ouml;lcskei et al.6, the procedures described here allows for the rapid, sensitive and reproducible determination of thermal nociceptive thresholds (TNTs) in mice and rats. The method uses slowly increasing thermal stimulus applied mostly to the skin of mouse/rat plantar surface. The method is particularly sensitive to study anti-nociception during hyperalgesic states such as PDN. The procedures described bellow are based on the ones published in detail by Alm&aacute;si et al 5 and B&ouml;lcskei et al 3. The procedures described here have been approved the Laboratory Animal Care and Use Committee (LACUC), Wright State University.

Watch this Scientific Journal Video about Rapid Determination of the Thermal Nociceptive Threshold in Diabetic Rats at JoVE.com

Rapid Determination of the Thermal Nociceptive Threshold in Diabetic Rats

Here, we describe a rapid reliable and simple procedure to determine the lowest temperature at which rats or mice show nocifensive behavior, i.e. the thermal nociceptive threshold (TNT). This method applies a slowly increasing thermal stimulus allowing precise and reproducible estimation of TNTs with minimum, if any, stress to the animals.

Painful diabetic neuropathy (PDN) is characterized by hyperalgesia i.e., increased sensitivity to noxious stimulus, and allodynia i.e., hypersensitivity ...

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