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This is particularly true for the field of pain research, where ethical standards are notably sensitive. The formalin test is widely used in pain research and some of its mechanisms resemble those underlying clinical pain in humans. Immediately upon injection, formalin triggers two waves (an early and a late phase) of strong, nociceptive behaviour, characterised by licking, biting, lifting and shaking the injected paw of the animal. Although well characterised at the behaviour level, since its proposal over four decades ago, there has not been any significant refinement to the formalin test, especially those combining minimisation of animal distress and preservation of behavioural outcomes of the test.\u00a0 Here, we propose a modified and improved method for the formalin test. We show that anaesthetising the animal with the inhalable anaesthetic sevoflurane at the time of the injection can produce reliable, robust and reproducible results whilst animal distress during the initial phase is reduced. Importantly, our results were validated by pharmacological suppression of the behaviour during the late phase of the test with gabapentin, the anaesthetic showing no interference with the drug. In addition, we demonstrate that this is also a useful method to screen for changes in pain behaviour in response to formalin in transgenic lines.<\/ns4:p>","DOI":"10.12688\/f1000research.18338.2","type":"journal-article","created":{"date-parts":[[2019,8,2]],"date-time":"2019-08-02T10:00:23Z","timestamp":1564740023000},"page":"891","update-policy":"https:\/\/doi.org\/10.12688\/f1000research.crossmark-policy","source":"Crossref","is-referenced-by-count":13,"title":["A refinement to the formalin test in mice"],"prefix":"10.12688","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1056-9199","authenticated-orcid":false,"given":"Douglas M","family":"Lopes","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heather L","family":"Cater","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew","family":"Thakur","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara","family":"Wells","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen B","family":"McMahon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"2560","published-online":{"date-parts":[[2019,8,2]]},"reference":[{"key":"ref-1","doi-asserted-by":"publisher","first-page":"161-74","DOI":"10.1016\/0304-3959(77)90130-0","article-title":"The formalin test: a quantitative study of the analgesic effects of morphine, meperidine, and brain stem stimulation in rats and cats.","volume":"4","author":"D Dubuisson","year":"1977","journal-title":"Pain."},{"key":"ref-2","doi-asserted-by":"publisher","first-page":"3671-5","DOI":"10.1523\/JNEUROSCI.12-09-03671.1992","article-title":"The role of NMDA receptor-operated calcium channels in persistent nociception after formalin-induced tissue injury.","volume":"12","author":"T Coderre","year":"1992","journal-title":"J Neurosci."},{"key":"ref-3","doi-asserted-by":"publisher","first-page":"37-42","DOI":"10.15412\/J.BCN.03080105","article-title":"Characterization of Nociceptive Behaviors Induced by Formalin in the Glabrous and Hairy Skin of Rats.","volume":"8","author":"E Erami","year":"2017","journal-title":"Basic Clin Neurosci."},{"key":"ref-4","doi-asserted-by":"publisher","first-page":"637-40","DOI":"10.1016\/0091-3057(81)90222-7","article-title":"Apparent lack of tolerance in the formalin test suggests different mechanisms for morphine analgesia in different types of pain.","volume":"15","author":"F Abbott","year":"1981","journal-title":"Pharmacol Biochem Behav."},{"key":"ref-5","doi-asserted-by":"publisher","first-page":"235-42","DOI":"10.1016\/0304-3959(90)91167-H","article-title":"The formalin test in mice: effect of formalin concentration.","volume":"42","author":"J Rosland","year":"1990","journal-title":"Pain."},{"key":"ref-6","doi-asserted-by":"publisher","first-page":"145-163","DOI":"10.3727\/000000003783992982","article-title":"The Formalin test: characteristics and usefulness of the model.","volume":"7","author":"J Sawynok","year":"2004","journal-title":"Rev Analg."},{"key":"ref-7","doi-asserted-by":"publisher","first-page":"5-17","DOI":"10.1016\/0304-3959(92)90003-T","article-title":"The formalin test: an evaluation of the method.","volume":"51","author":"A Tjolsen","year":"1992","journal-title":"Pain."},{"key":"ref-8","doi-asserted-by":"publisher","first-page":"247-54","DOI":"10.1016\/0304-3959(93)90220-J","article-title":"The tail-flick and formalin tests in rodents: changes in skin temperature as a confounding factor.","volume":"53","author":"K Hole","year":"1993","journal-title":"Pain."},{"key":"ref-9","doi-asserted-by":"publisher","first-page":"69-76","DOI":"10.1016\/0165-0270(85)90116-5","article-title":"Formalin test in mice, a useful technique for evaluating mild analgesics.","volume":"14","author":"S Hunskaar","year":"1985","journal-title":"J Neurosci Methods."},{"key":"ref-10","doi-asserted-by":"publisher","first-page":"103-14","DOI":"10.1016\/0304-3959(87)90088-1","article-title":"The formalin test in mice: dissociation between inflammatory and non-inflammatory pain.","volume":"30","author":"S Hunskaar","year":"1987","journal-title":"Pain."},{"key":"ref-11","doi-asserted-by":"publisher","first-page":"347-52","DOI":"10.1016\/0304-3959(89)90222-4","article-title":"Modified formalin test: characteristic biphasic pain response.","volume":"38","author":"M Shibata","year":"1989","journal-title":"Pain."},{"key":"ref-12","doi-asserted-by":"publisher","first-page":"175-86","DOI":"10.1016\/0160-5402(88)90078-2","article-title":"Methodological refinements to the mouse paw formalin test. 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Afify","year":"2017","journal-title":"Front Neurol."},{"key":"ref-40","doi-asserted-by":"publisher","first-page":"106-10","DOI":"10.1016\/0006-8993(92)90932-y","article-title":"Infusions of 6-hydroxydopamine into the nucleus accumbens abolish the analgesic effect of amphetamine but not of morphine in the formalin test.","volume":"580","author":"P Clarke","year":"1992","journal-title":"Brain Res."},{"key":"ref-41","doi-asserted-by":"publisher","first-page":"220-7","DOI":"10.1007\/PL00005345","article-title":"The inflammatory reaction induced by formalin in the rat paw.","volume":"359","author":"J Damas","year":"1999","journal-title":"Naunyn Schmiedebergs Arch Pharmacol."},{"key":"ref-42","doi-asserted-by":"publisher","first-page":"263-70","DOI":"10.1016\/S0304-3959(99)00212-2","article-title":"Opioid inhibition of formalin-induced changes in plasma extravasation and local blood flow in rats.","volume":"84","author":"B 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Sci."},{"key":"ref-55","doi-asserted-by":"publisher","first-page":"257-273","DOI":"10.1016\/j.neuron.2014.12.007","article-title":"The roles of PINK1, parkin, and mitochondrial fidelity in Parkinson's disease.","volume":"85","author":"A Pickrell","year":"2015","journal-title":"Neuron."},{"key":"ref-56","doi-asserted-by":"publisher","first-page":"936-946","DOI":"10.1038\/nprot.2006.120","article-title":"Housing, husbandry and handling of rodents for behavioral experiments.","volume":"1","author":"R Deacon","year":"2006","journal-title":"Nat Protoc."},{"key":"ref-57","doi-asserted-by":"publisher","first-page":"825-826","DOI":"10.1038\/nmeth.1500","article-title":"Taming anxiety in laboratory mice.","volume":"7","author":"J Hurst","year":"2010","journal-title":"Nat Methods."},{"key":"ref-58","doi-asserted-by":"publisher","first-page":"1089-1098","DOI":"10.1016\/j.neubiorev.2009.05.004","article-title":"Restraint stress in biobehavioral research: Recent developments.","volume":"33","author":"T Buynitsky","year":"2009","journal-title":"Neurosci Biobehav Rev."},{"key":"ref-59","doi-asserted-by":"publisher","first-page":"112-119","DOI":"10.1016\/j.pnpbp.2012.05.018","article-title":"Chronic restraint stress causes anxiety- and depression-like behaviors, downregulates glucocorticoid receptor expression, and attenuates glutamate release induced by brain-derived neurotrophic factor in the prefrontal cortex.","volume":"39","author":"S Chiba","year":"2012","journal-title":"Prog Neuropsychopharmacol Biol Psychiatry."},{"key":"ref-60","doi-asserted-by":"publisher","first-page":"299-305","DOI":"10.1006\/exnr.1996.0103","article-title":"Restraint reduces formalin-test pain but the effect is not influenced by lesions of the hypothalamic paraventricular nucleus.","volume":"139","author":"P Fuchs","year":"1996","journal-title":"Exp Neurol."},{"key":"ref-61","doi-asserted-by":"publisher","first-page":"174-182","DOI":"10.1016\/j.neuroscience.2007.04.012","article-title":"Opioid modulation of reflex versus operant responses following stress in the rat.","volume":"147","author":"C King","year":"2007","journal-title":"Neuroscience."},{"key":"ref-62","doi-asserted-by":"publisher","first-page":"214-222","DOI":"10.1016\/S0006-8993(03)03339-0","article-title":"Differential effects of stress on escape and reflex responses to nociceptive thermal stimuli in the rat.","volume":"987","author":"C King","year":"2003","journal-title":"Brain Res."},{"key":"ref-63","doi-asserted-by":"publisher","first-page":"643-649","DOI":"10.1016\/j.physbeh.2005.12.007","article-title":"Nociception- and anxiety-like behavior in rats submitted to different periods of restraint stress.","volume":"87","author":"G Gameiro","year":"2006","journal-title":"Physiol Behav."},{"key":"ref-64","doi-asserted-by":"publisher","first-page":"9-12","DOI":"10.1016\/0304-3940(95)12157-Y","article-title":"Exaggerated cardiovascular and behavioral nociceptive responses to subcutaneous formalin in the spontaneously hypertensive rat.","volume":"201","author":"B Taylor","year":"1995","journal-title":"Neurosci Lett."},{"key":"ref-65","first-page":"223-229","article-title":"Refinement of pain evaluation techniques. The formalin test.","volume":"40","author":"F Capone","year":"2004","journal-title":"Ann Ist Super Sanita."},{"key":"ref-66","doi-asserted-by":"publisher","first-page":"1300-1304, table of contents","DOI":"10.1097\/00000539-200211000-00037","article-title":"The analgesic effect of xenon on the formalin test in rats: a comparison with nitrous oxide.","volume":"95","author":"T Fukuda","year":"2002","journal-title":"Anesth Analg."},{"key":"ref-67","doi-asserted-by":"publisher","first-page":"S255","DOI":"10.1016\/0304-3959(87)91570-3","article-title":"Afferent activity in models of inflammation.","volume":"30","author":"N Russell","year":"1987","journal-title":"Pain."},{"key":"ref-68","doi-asserted-by":"publisher","first-page":"345-355","DOI":"10.1016\/0304-3959(95)00121-2","article-title":"Formalin-evoked activity in identified primary afferent fibers: systemic lidocaine suppresses phase-2 activity.","volume":"64","author":"S Puig","year":"1996","journal-title":"Pain."},{"key":"ref-69","doi-asserted-by":"publisher","first-page":"422-429","DOI":"10.1016\/j.pain.2010.08.001","article-title":"Pain behavior in the formalin test persists after ablation of the great majority of C-fiber nociceptors.","volume":"151","author":"S Shields","year":"2010","journal-title":"Pain."},{"key":"ref-70","doi-asserted-by":"publisher","first-page":"11412-11415","DOI":"10.1523\/JNEUROSCI.3600-07.2007","article-title":"An ion channel essential for sensing chemical damage.","volume":"27","author":"L Macpherson","year":"2007","journal-title":"J Neurosci."},{"key":"ref-71","doi-asserted-by":"publisher","first-page":"13525-13530","DOI":"10.1073\/pnas.0705924104","article-title":"TRPA1 mediates formalin-induced pain.","volume":"104","author":"C McNamara","year":"2007","journal-title":"Proc Natl Acad Sci U S 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serotonergic innervation of the olfactory bulb.","volume":"235","author":"L Glasl","year":"2012","journal-title":"Exp Neurol."},{"key":"ref-76","doi-asserted-by":"publisher","first-page":"1889-1901","DOI":"10.1242\/dev.00417","article-title":"Directional guidance of interneuron migration to the cerebral cortex relies on subcortical Slit1\/2-independent repulsion and cortical attraction.","volume":"130","author":"O Marin","year":"2003","journal-title":"Development."}],"container-title":["F1000Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/f1000research.com\/articles\/8-891\/v2\/xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/f1000research.com\/articles\/8-891\/v2\/pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/f1000research.com\/articles\/8-891\/v2\/iparadigms","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2019,9,21]],"date-time":"2019-09-21T02:46:10Z","timestamp":1569033970000},"score":1,"resource":{"primary":{"URL":"https:\/\/f1000research.com\/articles\/8-891\/v2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,8,2]]},"references-count":76,"URL":"https:\/\/doi.org\/10.12688\/f1000research.18338.2","relation":{"has-review":[{"id-type":"doi","id":"10.5256\/f1000research.22123.r52019","asserted-by":"subject"},{"id-type":"doi","id":"10.5256\/f1000research.22123.r52019","asserted-by":"object"}]},"ISSN":["2046-1402"],"issn-type":[{"value":"2046-1402","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,8,2]]},"assertion":[{"value":"Indexed","URL":"https:\/\/f1000research.com\/articles\/8-891\/v2#article-reports","order":0,"name":"referee-status","label":"Referee status","group":{"name":"current-referee-status","label":"Current Referee Status"}},{"value":"10.5256\/f1000research.22123.r52019, Nora Bourbia, Public Health England, Chilton, UK, 16 Aug 2019, version 2, indexed","URL":"https:\/\/f1000research.com\/articles\/8-891\/v2#referee-response-52019","order":0,"name":"referee-response-52019","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.20060.r50152, John Riddell, Andrew Todd, Spinal Cord Group, Institute of Neuroscience and Psychology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK, 22 Jul 2019, version 1, 1 approved, 1 approved with reservations","URL":"https:\/\/f1000research.com\/articles\/8-891\/v1#referee-response-50152","order":1,"name":"referee-response-50152","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"<b>Douglas Lopes<\/b>; \n<i>Posted: 30 Jul 2019<\/i>; \n<b>We would like to thank the reviewers for the very positive reviews of our paper. The reviewer\u2019s comments were very helpful, and we have now revised our manuscript to address all of the issues raised.<\/b>  \n<i>The results of the study are generally convincing, and the article is clearly written and well illustrated. One concern is the relatively high level of pain behaviour in the control group for the Pink1 study (Fig 4). While the area under the curve for other groups is typically around 1000, for the controls in Fig 4 it is nearly 2000. In fact, compared to other cohorts, the Pink1-\/- mice seem to have normal behaviour, while the control group show an exaggerated response. Some explanation is needed here. <\/i>&nbsp; \n<b>This point was was also raised by the other reviewer and addressed accordingly.&nbsp; We trust the behaviour observed represents the natural variation it can be obtained when performed behavioural tests and therefore we can only emphasise why it is important to control with animals that have been bred at the same time. Although we understand and acknowledge the reviewer\u2019s concerns, we trust the data is a true representation of the mice behaviour for this particular line. As demonstrated on the graph (Supplementary Figure 1 B), all the control animals used in this test were above the average control response in the other figures (above ~250 sec.), we therefore believe it is unlikely the findings are a false positive. We hypothesise that there is not a biological obvious reason that can explain this variation apart from the fact that they are a different batch of animals and thus likely to differ in experimenter, day and uncontrollable variances in their cage environment (such as genotype of parents, litters, litter size, etc.). Unfortunately, this line is no longer promptly available (it has been cryopreserved and archived) and therefore we are regrettably unable to repeat these experiments to investigate this further.&nbsp; We have however, added a sentence in the manuscript pointing out the differences observed and explaining it as above.<\/b> &nbsp; \n<i>Also \u2013 what are the units for area under curve?<\/i> \n<b>The AUC was calculated in relation to the pain response (sec) over time. This has been added to the materials and methods<\/b> &nbsp; \n<i>Minor points:<\/i> &nbsp; \n<i>It would be helpful to have some explanation of why the Pink1 and Slit1 mouse lines were chosen for this study, at the end of the Experimental Animals paragraph in the Methods section.<\/i> \n<b>These sentences were added: The transgenic lines chosen in this study were part of a parallel neuroscience program study being carried out at the MLC. In addition, it has been suggested a link between Pink1 and nociceptive processing (48,49) and Slit1 expression and peripheral injury (50\u201355). <\/b> &nbsp; \n<i>Page 4: In the 4th line of the Results section &quot;it is&quot; should be inserted before &quot;being&quot; \u2013 otherwise it would be the experimenter who received the injection.<\/i> \n<b>This has been amended.<\/b> &nbsp; \n<i>Page 5: left column, 6th line &quot;led&quot; rather than &quot;lead&quot;.<\/i> \n<b>This has been amended.<\/b> &nbsp; \n<i>Page 6: left column, 6th line \u2013 omit &quot;seem to&quot;.<\/i>\n<b> This has been amended.<\/b>","URL":"https:\/\/f1000research.com\/articles\/8-891\/v2#referee-comment-4788","order":2,"name":"referee-comment-4788","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"This work was supported by the Wellcome Trust [102645]; Medical Research Council [53658, 53650]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.","order":3,"name":"grant-information","label":"Grant Information"},{"value":"This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.","order":0,"name":"copyright-info","label":"Copyright"}]}}