Nociception assay

A nociception assay (nocioception or nocioperception assay) evaluates the ability of an animal, usually a rodent, to detect a noxious stimulus such as the feeling of pain, caused by stimulation of nociceptors. These assays measure the existence of pain through behaviors such as withdrawal, licking, immobility, and vocalization. The sensation of pain is not a unitary concept; therefore, a researcher must be conscious as to which nociception assay to use.

The formalin assay is the most popular chemical assay of nociception. It entails the injection of a dilute solution of formalin into the surface of the rodent's hindpaw, followed by the scoring of stereotypical behaviors such as flinching, licking, and biting of the affected hindpaw. The behaviors last for approximately 1 hour, with the early or acute stage (directly after injection) reflecting direct activation of nociceptors and the late or tonic phase (15 to 20 minutes after the injection) reflecting inflammation. Typically, the formalin assay is used on rats; however, formalin concentrations and scoring methods can be modified as to suit mice. One major advantage of the formalin assay over other models of inflammatory pain is the limited duration (approximately 1 hour) of the response. Additionally, as described before, this assay produces a response in two discrete stages, allowing researchers to model both acute and tonic pain using a single noxious chemical.

The Von Frey assay, introduced by Maximilian von Frey and modified by Weinstein, uses Von Frey hair or fibers, which are small pieces of nylon rod, approximately 50 mm in length, and of varying diameters, to test a rodent's sensitivity to a mechanical stimulus. It is unclear whether the process is really considered noxious versus simply annoying, so this assay is a test of mechanical nociception or simply mechanical sensibility. In this test, the animal stands on an elevated mesh platform, and the Von Frey hairs are inserted through the mesh to poke the animal’s hindpaw. Normal reactions for the animal include withdrawing or licking or shaking the paw, and possible vocalization, but these can depend on variability within the experiment. For example, the ventral surface of the hind paw is typically associated with lower withdrawal thresholds compared to the dorsal surface, and the exact force of the fiber is determined by its thickness. It is also important to note that thresholds usually are initially decreasing during successive tests, but do become stable after about 3 sessions. Algorithms such as up-down or Bruceton analysis are available to concentrate testing into the most dynamic part of the range, and subsequent curve fitting and parameter estimation can be similarly standardised. Alternatively, automated von Frey systems have recently been discovered that gradually increase the force of a single probe so that a researcher can observe when withdrawal responses occur.

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