{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T05:06:41Z","timestamp":1772168801009,"version":"3.50.1"},"update-to":[{"DOI":"10.12688\/f1000research.130474.1","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2024,7,3]],"date-time":"2024-07-03T00:00:00Z","timestamp":1719964800000}},{"DOI":"10.12688\/f1000research.130474.2","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2024,7,3]],"date-time":"2024-07-03T00:00:00Z","timestamp":1719964800000}},{"DOI":"10.12688\/f1000research.130474.3","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2024,7,3]],"date-time":"2024-07-03T00:00:00Z","timestamp":1719964800000}}],"reference-count":20,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2024,7,3]],"date-time":"2024-07-03T00:00:00Z","timestamp":1719964800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000066","name":"The National Institute of Environmental Health Sciences","doi-asserted-by":"crossref","award":["R35ES028366"],"award-info":[{"award-number":["R35ES028366"]}],"id":[{"id":"10.13039\/100000066","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000066","name":"The National Institute of Environmental Health Sciences","doi-asserted-by":"crossref","award":["T32ES007126"],"award-info":[{"award-number":["T32ES007126"]}],"id":[{"id":"10.13039\/100000066","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["f1000research.com"],"crossmark-restriction":false},"short-container-title":["F1000Res"],"abstract":"<ns3:p>\n                    Background The open field assay is used to study anxiety-related traits and anxiolytic drugs in rodents. This assay entails measuring locomotor activity and time spent in the center of a chamber that is maintained at ambient room temperature. However, the ambient temperature in most laboratories varies daily and seasonally and can differ between buildings. We sought to evaluate how varying ambient temperature and core body temperature (CBT) affected open field locomotor activity and center time of male wild-type (WT, C57BL\/6) and Transient Receptor Potential Subfamily M Member 8 (\n                    <ns3:italic>Trpm8<\/ns3:italic>\n                    ) knock-out (\n                    <ns3:italic>\n                      Trpm8\n                      <ns3:sup>-\/-<\/ns3:sup>\n                    <\/ns3:italic>\n                    ) mice. TRPM8 is an ion channel that detects cool temperatures and is activated by icilin. Methods Mice were placed in the open field at 4\u00b0C and 23\u00b0C for 1 hour. Distance traveled and time spent in the center were measured. Mice were injected with icilin, M8-B, diazepam, or saline, and changes in activity level were recorded. Results The cooling agent icilin increased CBT and profoundly reduced distance traveled and center time of WT mice relative to controls. Likewise, cooling the ambient temperature to 4\u00b0C reduced distance traveled and center time of WT mice relative to\n                    <ns3:italic>\n                      Trpm8\n                      <ns3:sup>-\/-<\/ns3:sup>\n                    <\/ns3:italic>\n                    mice. Conversely, the TRPM8 antagonist (M8-B) reduced CBT and increased distance traveled and center time of WT mice when tested at 4\u00b0C. The TRPM8 antagonist (M8-B) had no effect on CBT or open field behavior of\n                    <ns3:italic>\n                      Trpm8\n                      <ns3:sup>-\/-<\/ns3:sup>\n                    <\/ns3:italic>\n                    mice. The anxiolytic diazepam reduced CBT in WT and\n                    <ns3:italic>\n                      Trpm8\n                      <ns3:sup>-\/-<\/ns3:sup>\n                    <\/ns3:italic>\n                    mice. When tested at 4\u00b0C, diazepam increased distance traveled and center time in WT mice but did not alter open field behavior of\n                    <ns3:italic>\n                      Trpm8\n                      <ns3:sup>-\/-<\/ns3:sup>\n                    <\/ns3:italic>\n                    mice. Conclusions Environmental temperature and drugs that affect CBT can influence locomotor behavior and center time in the open field assay, highlighting temperature (ambient and core) as sources of environmental and physiologic variability in this commonly used behavioral assay.\n                  <\/ns3:p>","DOI":"10.12688\/f1000research.130474.4","type":"journal-article","created":{"date-parts":[[2024,7,11]],"date-time":"2024-07-11T10:59:00Z","timestamp":1720695540000},"page":"234","update-policy":"https:\/\/doi.org\/10.12688\/f1000research.crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["The open field assay is influenced by room temperature and by drugs that affect core body temperature"],"prefix":"10.12688","volume":"12","author":[{"given":"Jessica A.","family":"Jimenez","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eric S.","family":"McCoy","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David F.","family":"Lee","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0911-7902","authenticated-orcid":false,"given":"Mark J.","family":"Zylka","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"2560","published-online":{"date-parts":[[2024,7,3]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"175-183","DOI":"10.1016\/j.vascn.2013.05.003","article-title":"Animal models of anxiety: a comprehensive review.","volume":"68","author":"V Kumar","year":"2013","journal-title":"J. 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In addition, the authors&nbsp;pharmacologically inhibited or activated the cold-sensing TRPM-8 cation receptors, in mice exposed at cold ambient temperature (4&deg;C), and described the interactions between TRPM-8 channel and cold exposure, as observed in the open field test.\u2028While is quite an interesting, I would like to encourage the authors to address the following comments:  1) In the main text the open field test lasted for 1 hours. However in the abstract it is mentioned 'Mice were placed in the open field at 4&deg;C and 23&deg;C for 30 minutes'. \n<b>Ans:&nbsp;<\/b>The open field was run for 1 hour. The abstract method was corrected.  2) Did the authors use male or female mice? And why?&nbsp;\n<u>It should be always clearly stated<\/u>. \n<b>Ans:&nbsp;<\/b>As stated in the Abstract Background, male WT and TRPM8 knock-out mice were used. This is now restated in the methods and discussion as a limitation. \u201cFor consistency, only male mice were used in this study. However, it should be stated that female mice may display different drug-induced behavioral responses and should be explored in future studies.\u201d  3) Can the authors explain the rationale for the doses tested for all compounds, as well as include references with the effects on these compounds in the open field test? \n<b>Ans:&nbsp;<\/b>This comment was addressed in the previous revision with appropriate reference \u201cHere we selected drug doses that are commonly used and well tolerated by mice in the behavior field.\u201d Almeida et al. used 6 mg\/kg i.p. M8-B to evaluate the hypothermic action of the drug. We used 12 mg\/kg as we didn\u2019t see an effect with 6 mg\/kg. Vinkers et al. evaluated stress-induced hyperthermia following 1-4 mg\/kg diazepam. Padua-Reis et al. also evaluated the anxiolytic effect of 2 mg\/kg diazepam. We used 2 mg\/kg. \n<i>New reference:<\/i> Colburn et al. evaluated icilin-induced jumping and cold sensation following 60 mg\/kg icilin administration (new reference). We used 50 mg\/kg.  4) The authors are rather measuring the skin temperature and not core body temperature. Can they please explain? They should also discuss how skin temperature correlates with CBT. \n<b>Ans:&nbsp;<\/b>There are also drugs that can solely influence the skin temperature. It should be noted. We did not measure skin temperature. CBT was assessed by measuring rectal temperature in mice, a technique that has been widely used for many years. Limitations of this procedure include distress to the animals, which could increase core temperature. However, however this risk was reduced by acclimating the mice to the procedure 2x each day for one week prior to testing. This was added to the methods.\u2028\u2028  5) In figure 2, Icilin alter skin temperature&nbsp; 90 mins post-injection. However, the open field starts 10 min after the injection and lasts for 60 mins (i.e. 70 mins post-injection).\u2028How, then, they concluded that altered temperature influences the open field behavior. Can they please explain? \n<b>Ans:&nbsp;<\/b>Previous studies had already shown a rapid change in core body temperature post-icilin injection using thermal telemetry and at doses lower than the one used in the current study, and these significant changes in CBT lasted at least 90 minutes (Knowlton et al, 2011 \u2013 \n<i>New Reference<\/i>). Using a less sensitive device, we did see an increase in CBT during the open field studies consistent with these studies. We likely did not see a change in CBT earlier due to our low sample size and less sensitive temperature detection method. This information was added to the results section.  6) In figure 6, it should be noted that no vehicle-injected mice were used. It is common that skin temperature increases and decreases rapidly upon i.p injections. Without an appropriate control experiments, it is rather difficult to confidently conclude whether diazepam, indeed, reduces body temperature. That is certainly a limitation and should be noted. \n<b>Ans:&nbsp;<\/b>To consider temperature changes upon i.p. injections, the results from figures 4 and 6 are compared with their saline-injected vehicle controls in figure 5. Diazepam was dissolved in saline and M8-B was dissolved DMSO. This is included in the methods section. The following limitation is included in the discussion as a previous revision: \u201cOf note, our vehicle controls in Figure 5 were saline injected, which may limit the interpretation of this study.\u201d However, \u201cDMSO at the concentration used to dissolve Icilin and M8-B have also been reported to be well tolerated by mice\u201d.\u2028\u2028  7) Its extremely unclear what is the difference of figure 6 and 3. In this graph it seems that&nbsp; diazepam has no effect at all.\u2028The authors should explain and also further discuss these results in conclusion. \n<b>Ans:&nbsp;<\/b>Figure 6 represents the effect of diazepam on CBT and open field behavior in WT and Trpm8 knock out mice. Panel A shows the effect of diazepam on the core body temperature. Panel B shows the effect of diazepam on the open field behavior of these two genotypes at 4C. Figure 3 represents the effect of temperature on the open field behavior of WT and Trpm8 knock out mice. Mice are not injected with drugs in this experiment. Both genotypes are assessed in the open field at room temperature and in the cold room at 4C. A new graph was included in the previous revision to better represent this experiment.","URL":"https:\/\/f1000research.com\/articles\/12-234\/v4#referee-comment-11911","order":2,"name":"referee-comment-11911","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.162395.r274455, Kei Nagashima, Body Temperature and Fluid Laboratory, Graduate School of Human Sciences, Faculty of Human Sciences, Waseda University, Saitama, 10 Jun 2024, version 3, indexed","URL":"https:\/\/f1000research.com\/articles\/12-234\/v3#referee-response-274455","order":3,"name":"referee-response-274455","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"This work was supported by grants to M.J.Z. from The National Institute of Environmental Health Sciences (NIEHS; R35ES028366). J.A.J. was supported by the Curriculum in Toxicology and Environmental Medicine Training Grant (NIEHS, T32ES007126). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.","order":4,"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"}]}}