使用电子Von Frey测痛仪评估大鼠的机械痛觉超敏

Jeremy Ferrier; Fabien Marchand; David Balayssac

doi:10.21769/BioProtoc.1933

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Peer-reviewed

Assessment of Mechanical Allodynia in Rats Using the Electronic Von Frey Test

使用电子Von Frey测痛仪评估大鼠的机械痛觉超敏

Jeremy Ferrier ^*

Fabien Marchand ^*

David Balayssac email

(*contributed equally to this work) 发布: 2016年09月20日第6卷第18期 DOI: 10.21769/BioProtoc.1933 浏览次数: 27533

评审: Oneil G. BhalalaAntoine de MorreeAnonymous reviewer(s)

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参见作者原研究论文

The authors used this protocol in:

Cover of The Journal of Neuroscience, featuring study using the protocol.

Dec 2015

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Abstract

Chronic pain is one of the most debilitating conditions, affecting one out of five people worldwide. Preclinical models in rodents represent a valuable tool for the study of pathophysiological mechanisms and the discovery of new analgesic drugs. However, pain evaluation in rodents is rather challenging. Altered response to mechanical or thermal stimuli is commonly used as behavioral outcome to measure pain sensitivity. This protocol introduces a method for assessing static mechanical pain hypersensitivity in rats using an electronic von Frey (VF) test. The electronic VF is an evolution of the manual VF hairs previously described by Kim and Chung (1991). In this previous procedure, 6 calibrated nylon filaments (diameters of 0.13, 0.23, 0.31, 0.48, 0.52, and 0.59 mm, respectively) are perpendicularly applied to the plantar surface of the rat hind paw, to deliver the following defined pressures: 5.89, 9.81, 27.0, 74.4, 124, and 205 mN. Each application has to be repeated several times for each filament to determine the mechanical threshold. Comparatively, the electronic VF is relatively easy-to-use and particularly suited for pharmacological studies with precise time-points. The electronic VF can be used as a behavioural read-out for a wide range of models, including inflammatory and neuropathic pain. The following protocol was originally published in Ferrier et al. (2013), Grégoire et al. (2014) and in Ferrier et al. (2015).

Keywords: Electronic von frey (电子Von Frey)

Materials and Reagents

Plastic tip (Harvard Apparatus, catalog number: 76-0488 )
Note: The tip doesn’t need to be changed between rats and between sessions as long as it is not damaged or dirty beyond cleaning.
Rats
Note: To reach a sufficient statistical power, use a minimum of 8 rats per group. Use preferentially Sprague-Dawley male rats, weighing 200 to 400 g (6 to 12 weeks old), since this protocol has only been validated on this strain of rats. However, there are no obvious reasons that this protocol won’t be suitable for others strains of rats.
Cleaning solution
Note: Within one session, the boxes are carefully cleaned with water to remove the odor of the previous animal. At the end of the session, clean the boxes using 70% ethanol.
Oxaliplatin (6 mg/kg, dissolved in 5% glucose)

Equipment

Apparatus (either home-made or commercially available at Harvard Apparatus, catalog number: 76-0886/87 )
Note: The apparatus consists of an elevated horizontal wire mesh stand (Figure 1). The dimensions of the Plexiglas boxes set on top of the wire grid are 17 x 11 x 13 cm. The boxes are covered with perforated lids. Frosted or opaque separators can be used to diminish interactions between rats.

Figure 1. Apparatus for the electronic von Frey test
Electronic von Frey test (BIO-EVF4, Bioseb^®, France)
Note: The apparatus consists of a portable force transducer fitted with a plastic tip (Figure 2). When applying a pressure on the tip, the maximal force applied (in grams) is automatically recorded by the electronic device and displayed on the screen. The maximal pressure (cut-off) will be reached when the paw of the animal is lifted by the plastic tip. Between two stimulations (separated by at least 3 min), the force value is set to zero simply by pushing a foot pedal.

Figure 2. Electronic von Frey test device

Procedure

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Readers should cite both the Bio-protocol article and the original research article where this protocol was used:

Ferrier, J., Marchand, F. and Balayssac, D. (2016). Assessment of Mechanical Allodynia in Rats Using the Electronic Von Frey Test. Bio-protocol 6(18): e1933. DOI: 10.21769/BioProtoc.1933.
Ferrier, J., Bayet-Robert, M., Dalmann, R., El Guerrab, A., Aissouni, Y., Graveron-Demilly, D., Chalus, M., Pinguet, J., Eschalier, A., Richard, D., Daulhac, L., Marchand, F. and Balayssac, D. (2015). Cholinergic neurotransmission in the posterior insular cortex is altered in preclinical models of neuropathic pain: key role of muscarinic M2 receptors in donepezil-induced antinociception. J Neurosci 35(50): 16418-16430.