The Behavioral Spectrometer has been validated in a mouse model of carrageenan-induced hind paw inflammation, where the frequency of grooming was increased and the number of ambulations were decreased (Brodkin et al., 2014). J. doi: 10.1016/s1063-4584(03)00163-8, Bradman, M. J., Ferrini, F., Salio, C., and Merighi, A. It is descended from wild Norway rats, Rattus norvegicus, which despite their name likely originated in Asia. Woolfe, G., and Macdonald, A. D. (1944). The advantage of this approach is that each animal receives the same number and type of stimuli, although the number of tests per hind paw could exceed 50 (e.g., five different von Frey filaments each applied 10 times), which is not only time consuming, but potentially exposes animals with mechanical hind paw sensitivity to a disproportionate number of more aversive stimuli. Sci. 372, 301312. Pain 9, 445452. This therefore provides measure of paw withdrawal threshold on a continual scale, as the force is applied continuously and not in steps. Lancet 353, 19591964. The clinical picture of neuropathic pain. Two centuries ago, the brown rat Rattus norvegicus was used by scientists to understand human physiology and medicine, concentrating on the effects of food and oxygen deprivation. Irwin, S., Houde, R. W., Bennett, D. R., Hendershot, L. C., and Seevers, M. H. (1951). Rats were the first mammalian species specifically domesticated to be used in the laboratory. Lab. Animals 6:47. doi: 10.3390/ani6080047. | Terms | Privacy Policy, Rodent Hypoxia-Ischemia Models for Cerebral Palsy Research: A Systematic Review, The influence of immaturity on hypoxic-ischemic brain damage in the rat. (2013). Nonetheless, the thermal probe test remains to be validated in other pain models that cause more pronounced mechanical allodynia. Ann. doi: 10.1016/0304-3959(92)90041-9, Knowlton, W. M., Bifolck-Fisher, A., Bautista, D. M., and McKemy, D. D. (2010). However, in practice it is difficult to distinguish between allodynia and hyperalgesia in animals, and the terms allodynia and hyperalgesia are often used incorrectly or interchangeably in the literature. Here, the rodent is placed on the plate after it has been cooled to the desired temperature and the time taken to evoke nociceptive behavior such as shaking, jumping or licking in the animal is recorded as the response time. doi: 10.1021/je00019a016, Viscardi, A. V., Hunniford, M., Lawlis, P., Leach, M., and Turner, P. V. (2017). Gait analysis of C57BL/6 mice with complete Freunds adjuvant-induced arthritis using the CatWalk system. Effects of morphine on thermal sensitivity in adult and aged rats. Quantitative sensory testing in the german research network on neuropathic pain (DFNS): standardized protocol and reference values. Life Sci. The term nociception was coined by Charles Sherrington in the early 1900s to distinguish the sensation of paina result of central nervous system processingfrom the physiological phenomenon of the peripheral nervous system responding to potential harmful stimuli (Dubner, 1983; Coutaux et al., 2005). The Randall-Selitto or paw pressure test was developed as a tool to assess response thresholds to mechanical pressure stimulation and is often considered a measure of mechanical hyperalgesia (Figure 1C; Randall and Selitto, 1957). Lett. The Wistar outbred albino rat is perhaps the most popular rat model due to its multi-purpose characteristics, making it a powerful species for medical research into infectious diseases, or as a surgical model. Development of a piglet grimace scale to evaluate piglet pain using facial expressions following castration and tail docking: a pilot study. This test is also referred to as the two-temperature choice assay or thermal place preference test and can be used to assess both cold or heat avoidance or preference (Moqrich et al., 2005). Sensory determinants of thermal pain. (2017b). doi: 10.1016/S0304-3959(01)00437-7, Menndez, L., Lastra, A., Hidalgo, A., and Baamonde, A. The withdrawal response is detected visually by the researcher, resulting in subjective measurement of the threshold. Simmons is a student in the Kenan-Flagler Business School at the University of North Carolina at Chapel Hill. While weight bearing is typically considered to be a measure of non-stimulus evoked nociception, it can be argued that ambulation itself applies a nociceptive mechanical stimulus to the affected limb(s), and it may therefore be a measure of stimulus-evoked nociceptive behavior, especially in the dynamic weight bearing test or gait analysis tests. A radiant or infrared heat source is positioned underneath the animal and aimed at the plantar surface of the hind paw (Figure 3C). Rodents are allowed to freely move between a reference plate (neutral temperature) and test plate. Role of the NLRP3 inflammasome in a model of acute burn-induced pain. Both the mouse and rat grimace scale are limited by the need for extensive training of the observer and a degree of subjectivity, which could lead to variability. Assoc. Cartil. The exception is use of the test on the tail of mice (Minett et al., 2014), although this may not be useful to assess nociceptive behaviors in commonly used models that are localized to the hind paw. doi: 10.1016/S0014-2999(01)01302-4, Jensen, T. S., and Yaksh, T. L. (1986). Eur. (2014). February 8, 2018| Kristen Coughlin | CRISPR, Rat Models, Last Updated on January 23, 2023 by ingenious. Mol. Front. (2011). doi: 10.1002/9780470942390.mo110116, Mogil, J. S. (2009). The device automatically records the temperature that paw withdrawal occurred (in this case 50C). The burrowing assay has been validated for models of post-surgical pain in mice and in models of peripheral nerve injury, osteoarthritis and inflammation induced by CFA in rats (Jirkof et al., 2010; Andrews et al., 2012; Bryden et al., 2015). J. Pain 3:40. doi: 10.1186/1744-8069-3-40, Pitcher, G. M., Ritchie, J., and Henry, J. L. (1999). However, concerns over the clinical translatability of stimulus-evoked nociception in recent years has led to the development and increasing implementation of non-stimulus evoked methods, such as grimace scales, burrowing, weight bearing and gait analysis. TRPM8, but not TRPA1, is required for neural and behavioral responses to acute noxious cold temperatures and cold-mimetics in vivo. Adams, B. L., Guo, W., Gors, R. T., and Knopp, K. L. (2016). Osteoarthr. From a physiological standpoint, more is known about the responses and pathways in rats than in other species due to the wealth of data collected over the years. A notable shift has taken place over the last two decades, with mice taking a more and more prominent role in biomedical science compared to rats. Paw withdrawal threshold in the von Frey hair test is influenced by the surface on which the rat stands. Rodent Hypoxia-Ischemia Models for Cerebral Palsy Research: A Systematic Review. Am. The rodent is placed in an inclined holder with the hind paws resting on two separate pressure sensors. A modified version of the thermal probe test suitable for quantifying heat thresholds in rats is still to be developed. doi: 10.1016/0304-3959(88)90026-7, Harvey, V. L., and Dickenson, A. H. (2009). It also demonstrates two additional concepts, originally outlined by Pavlov. A., Possani, L. D., Cabot, P. J., Lewis, R. J., et al. Efficacy of drugs with different mechanisms of action in relieving spontaneous pain at rest and during movement in a rat model of osteoarthritis. In fact, it is a requirement for publication of in vivo data in high quality journals that relevant standards and guidelines are strictly adhered to (McGrath et al., 2010). Pain 50, 355363. Pain 12:1744806916665366. doi: 10.1177/1744806916665366, Yoburn, B. C., Morales, R., Kelly, D. D., and Inturrisi, C. E. (1984). What are some disadvantages of using Wistar rats in research and some disadvantages in using rats over mice in general? The hot plate test, first described in 1944, can be used to determine heat thresholds in mice and rats (Woolfe and Macdonald, 1944). There are protections in place for the animals. - Quora Answer: I'm going to answer this purely from the point of view of toxicology studies, because that's where my experience is. 13, 587599. The tail flick test, first described in 1941, involves application of a heat stimulus to the tail of mice and rats, and the time taken for the tail to flick or twitch is recorded (DAmour and Smith, 1941; Figure 3A). Exp. doi: 10.1093/brain/awf055, Defrin, R., Shachal-Shiffer, M., Hadgadg, M., and Peretz, C. (2006). Eur. J. Neurosci. The thermal probe test: a novel behavioral assay to quantify thermal paw withdrawal thresholds in mice. As the nocifensive response can be too fast for an investigator to quantify in real time, video recordings that are played back in slow motion may be required to accurately analyze the response to acetone. 7. Methods 255, 92103. Figure 3.Methods used to assess heat-evoked pain like behaviors in rodents. Res. Similarly, measures to improve data quality, including appropriate randomization and blinding procedures, are key to ensure validity of the obtained results. The pressure is then increased at a constant rate until a nociceptive behavioral response is observed. Rev. First, the response to a specific temperature (typically 5C to 15C) can be recorded (Allchorne et al., 2005). Front. For example, if you call 100 people between 2 and 5 p.m. and ask whether they feel that they have enough free time in their daily schedule, most of the respondents might say "yes." In terms of the diseases that they are used to study, rats are usually not the model of choice when it comes to disorders related to neoplasms, blood and immune function. 47, 810. Rats are an ideal choice for many labs due to their physiological similarity to humans. While this is generally not an issue when testing the anti-nociceptive effects of compounds delivered systematically or when phenotyping transgenic mice, this may confound the results for unilateral models of pain or for compounds administered by intraplantar injection. Exceptionally adaptable, these rodents now inhabit almost all environments on Earth, especially near human settlements where they are often seen as pests. 37, 18351841. doi: 10.1016/s0165-0270(99)00004-7, Plone, M. A., Emerich, D. F., and Lindner, M. D. (1996). Readership. A camera below records the paw prints, which are illuminated by internally reflected light in the glass. Lancet Neurol. These measures can have a profound effect on the apparent anti-nociceptive efficacy of test compounds and should thus be carefully considered during experimental design (Winter and Flataker, 1965). 5, 94100. The first (1880-1950) was when mice and rats became common research animals. Rep. 7:40883. doi: 10.1038/srep40883, Deuis, J. R., Yin, K., Cooper, M. A., Schroder, K., and Vetter, I. Sci. Pain Med. The mouse genome is very similar to our own, making mouse genetic research particularly useful for the study of human diseases. Mol. 24, 19281939. While forepaw withdrawal often occurs first, hind paw withdrawal or licking is considered to be a more reliable indicator of nociception, as the forepaws are frequently used in grooming and exploration and are not consistently in contact with the metal surface (Woolfe and Macdonald, 1944; Minett et al., 2011). Sci. For instance, improving data homogeneity and enhancing statistical power (Dell et al., 2002) will result in fewer animals being necessary to achieve the required confidence level (Festing and Altman, 2002). doi: 10.1016/j.ejpain.2004.10.006, Wibbenmeyer, L., Sevier, A., Liao, J., Williams, I., Latenser, B., Lewis, R., et al. Rev. Methods 219, 4151. Neuropathic pain: aetiology, symptoms, mechanisms, and management. The Hargreaves test is usually carried out using a glass bottom enclosure, which can be heated to minimize errors arising from heat sink effects. The Advanced Dynamic Weight Bearing apparatus (Bioseb) was developed as a modification from static weight bearing or incapacitance tests and computes weight bearing for each of the front and rear paws, weight ratio and paw surface area in freely moving animals (Griffioen et al., 2015). Berryman, E. R., Harris, R. L., Moalli, M., and Bagi, C. M. (2009). Xing, H., Chen, M., Ling, J., Tan, W., and Gu, J. G. (2007). However, theyre more widely used in studying cardiovascular and metabolic disorders, as well as the onset of conditions that relate to the brain, digestive system, and various health problems related to behavior. This review article provides an overview, as well as discussion of the advantages and disadvantages of the most commonly used behavioral methods of stimulus-evoked and non-stimulus-evoked nociception used in rodents. A monofilament is applied perpendicularly to the plantar surface of the hind paw until it buckles, delivering a constant pre-determined force (typically 0.213.7 mN for mice and 5.998 mN for rats) for 25 s (Figure 1A). A rat model of bone cancer pain. (2010). Neurosci. Mouse model: Pros & Cons. "Its brain is larger than that of a mouse, so it's easier to study the structures," said Joyce. Figure 6.Methods used to assess non-stimulus evoked pain behaviors in rodents. Different methodological approaches are used to determine mechanical sensitivity using manual Von Frey, including the up-down, ascending stimulus or percent response method, all of which will be discussed below. doi: 10.1016/j.jbspin.2004.01.010, Cox, J. J., Reimann, F., Nicholas, A. K., Thornton, G., Roberts, E., Springell, K., et al. The Hargreaves test permits measurement of ipsilateral and contralateral heat thresholds, allowing each animal to serve as its own internal control in unilateral pain models. 72, 7479. The Rat Grimace Scale is also scored 02 depending on observed changes in facial features, and evaluates the extent of orbital tightening, nose/cheek flattening, ear change and whisker change (Sotocinal et al., 2011). Assessment of thermal sensitivity in rats using the thermal place preference test: description and application in the study of oxaliplatin-induced acute thermal hypersensitivity and inflammatory pain models. Figure 2.Visual representation of the different methodological approaches used to determine mechanical sensitivity using manual Von Frey. The mouse with a human ear on its back may have been one of the more bizarre and visually unsettling experiments carried out on a rodent, but mice have been used for scientific experiments since. In addition, care should be taken to design experiments that minimize distress and suffering. Neurosci. This test involved application of an increasing mechanical force to the surface of the paw or tail until withdrawal or vocalization occurs. Therefore grimace scales have the potential to monitor and improve the welfare of animals used not only in research, but also in farming and industry. Lab. Lab. [ 120] A soft transducer ensures minimal vibration and reduces touch-on responses. The test is carried out on mesh floor and acetone is dabbed or sprayed on the plantar surface of the hind paw (Figure 4A), eliciting cooling of the skin to innocuous temperatures of 1521C (Colburn et al., 2007; Leith et al., 2010), although the actual temperature varies with ambient temperature, skin temperature, and the amount of acetone applied. Ear position denotes the ears being pulled back and apart from their standard position (may feature vertical ridges). Monkeys, rabbits, cats, ferrets, pigs, sheep and chimpanzees are just some of the animals used for biomedical experiments, science education, and product and cosmetics testing. Joint Bone Spine 72, 359371. Importantly, the pain appears to be spontaneous, with no identifiable stimulus. Differentiating thermal allodynia and hyperalgesia using dynamic hot and cold plate in rodents. Mice are cost effective because they are cheap and easy to look after. Nose bulge refers to the bulge noticeable on the bridge of the nose, whereas cheek bulge refers to the rounded projection of the cheek muscle compared to its typical appearance. J. Pharmacol. This sample - and the results - are biased, as most workers are at their jobs during these hours. J. Neurosci. Key Takeaways. doi: 10.1097/NNR.0000000000000082, Gritsch, S., Bali, K. K., Kuner, R., and Vardeh, D. (2016). (2005). Using the Mouse Grimace Scale to reevaluate the efficacy of postoperative analgesics in laboratory mice. The up-down method is practically limited by commercial filaments that are not equally spaced (therefore, the average increment is used for d), incorrect labeling of the force in log units, and the need for the first filament to be close to the mean threshold (which may be unknown; Bradman et al., 2015). The main advantage of the thermal probe test is that the mice are placed in individual runs standing on bars instead of glass enabling access to the plantar surface, which allows simultaneous assessment of mechanical thresholds by von Frey, removing the need for acclimation in two different enclosures. J. Pharmacol. PLoS One 9:e98856. In this experiment, a previously unafraid baby was conditioned to become afraid of a rat. (2012). However, rodents do not vocalize in the audible range unless the pain is severe, making use vocalization as an endpoint ethically limited (Mogil, 2009). In line with this, the uptake of grimace scales to study pain in the scientific community has been low, with approximately 30 studies published using the method in rodents since its introduction by Langford et al. Clin. Mice are placed individually in small cages with a barred floor. In this test, the unrestrained mouse or rat is placed on a metal surface starting at a non-noxious temperature (<42C), and the temperature is increased at a constant rate until a nocifensive behavior is observed. However, this also requires animals to be acclimatized to the cages, to ensure ambulation and exploratory behaviors, which could be misinterpreted as a positive response, are kept to a minimum. In the past, housing of rodents used for research was designed primarily to prevent A disadvantage of this method is that the number of measurements per animal is variable and that it requires repeated, time-intensive measurements, which may lead to sensitization or learnt responses. Burn Care Res. With a much less expensive and more precise method for targeting genomes via embryos, the potential for rats as a model system of human disease could be pursued even easier. Rev. Wistar rats are big rats. An estimated 71% of all NIH grant applications from 2008 to 2015 mentioned the keywords mus musculus, mouse, or mice. Therefore, there is an urgent need to increase our understanding of the underlying mechanisms of pain and to develop new treatments. Practical mechanical threshold estimation in rodents using von Frey hairs/Semmes-Weinstein monofilaments: towards a rational method. PLoS One 9:e104458. While the underlying protocols and principles are different, it remains to be determined if electronic Von Frey activates a different subset of sensory neurons compared to manual Von Frey (e.g., high-threshold vs. low threshold mechanoreceptors). Mechanical pressure is applied focally to the dorsal or plantar surface of the hind paw or tail, which is placed between a pointed probe tip and a flat surface. FACTS and MYTHS - Animal Research at Stanford Rice JE 3rd, Vannucci RC, Brierley JB. Why are Rats Used in Research? | Kent Scientific Guidelines for reporting experiments involving animals: the ARRIVE guidelines. This continues until at least four readings are obtained after the first change of direction. J. Pharmacol. The most common case of bias is a result of non-response. TRPM8 mechanism of cold allodynia after chronic nerve injury. In 2001 however with the development of targeted genetic manipulation in mouse embryonic stem (ES) cells, rats took somewhat of a back seat as the rodent of choice for specific gene targeting because this technology was not yet available in the rat. However, it is proving useful as a tool to monitor animal welfare, not only in rodents, but also in other species, with grimace scales being developed for pigs, sheep and horses (Matsumiya et al., 2012; Miller and Leach, 2015; Dalla Costa et al., 2016; Hager et al., 2017; Viscardi et al., 2017). PLoS One 10:e0125822. The thermal probe test (MouseMet Thermal, Topcat Metrology) is a novel method recently described to quantify heat thresholds in mice (Deuis and Vetter, 2016). doi: 10.1097/fbp.0000000000000026, Banik, R. K., and Kabadi, R. A. 27, 1368013690. J. These decades of work will enable the integration of new techniques and technologies for cerebral palsy research and maximize the utility of the hypoxia-ischemia model. The rats ability to learn, remember, and interact gives them an advantage over other species, allowing for insights into the basic principles behind learning and behavior which can then be translated to human behavior and learning. In the cold plantar assay, the animal is placed in an enclosure with a clean glass floor. Dynamic weight bearing is an efficient and predictable method for evaluation of arthritic nociception and its pathophysiological mechanisms in mice. Alternatively, a continuous temperature gradienteither in linear (Figure 5B) or circular formcan be used to determine the preferred temperature in freely moving animals (Moqrich et al., 2005; Touska et al., 2016). This can be achieved through a variety of techniques. doi: 10.1016/s1474-4422(14)70102-4, Jensen, T. S., Gottrup, H., Sindrup, S. H., and Bach, F. W. (2001). The dynamic hot plate test, first described in 1984, uses an increasing temperature ramp rather than a constant temperature. 39, 18811890. Osteoarthr. 3. Cooling of the glass leads to unilateral exposure of the hind paws to a cooled surface, the temperature of which can be determined by attaching a thermocouple probe to the glass or skin. Behavioral and physiological methods for early quantitative assessment of spinal cord injury and prognosis in rats. Methods Mol. 95% of all animals used in research are rats because their genetic, biological and behavioral characteristics are so similar to that of humans. (B) Hot plate test. Voluntary response bias is another disadvantage that comes with a small sample size. Nociceptive responses to high and low rates of noxious cutaneous heating are mediated by different nociceptors in the rat: electrophysiological evidence. Ther. (B) The ascending stimulus method. Osteoarthr. The evaluation of the analgesic action of pethidine hydrocholoride (Demerol).