Pain Research: Methods and Protocols by Z. David Luo

By Z. David Luo

A state-of-the-art number of diversified effortlessly reproducible thoughts for dissecting the molecular mechanisms of ache transduction. those equipment hire a number of multidisciplinary ways starting from animal soreness versions and unmarried neutron choice to in vitro single-cell mRNA amplification. the gathering contains not just typical and state of the art equipment, but additionally novel options just recently utilized to discomfort study. The protocols stick to the profitable equipment in Molecular Biology™ sequence layout, each supplying step by step laboratory directions, an creation outlining the main in the back of the process, lists of kit and reagents, and pointers on troubleshooting and heading off identified pitfalls.

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If an extensive manipulation of the L4 spinal nerve is required to separate it from the L5 spinal nerve, it may be safer to discard the animal rather than taking a chance of damaging the L4 nerve. Once enough length of the L5 spinal nerve is freed from the adjacent structure, a piece of 6-0 silk thread is placed around the L5 spinal nerve and the nerve is tightly ligated to interrupt all axons in the nerve (see Note 10). Another option would be to cut the spinal nerve just distal to the ligation to make sure all fibers are interrupted.

Experimental drugs are usually given intrathecally 5 min prior to injury induction for pretreatments or 30 min after injury for posttreatment. 30 Allen and Yaksh Fig. 2. Representative graph of typical flinching profiles as measured per minute with an automated detector. Fig. 3. Diagram of injury area and of primary and secondary hyperalgesia testing. The extent of injury is noted by the hatched area of a hind paw. Mechanical withdrawal thresholds (secondary hyperalgesia) are tested in the area noted by the X, whereas thermal thresholds (primary hyperalgesia) are tested in the area noted by the circle.

Method 1. 25 mL) for rats and mice. 2. Dissect the skin and muscle using a no. 11 scalpel blade. 3. Expose the sciatic nerve at midthigh level with the aid of wound expander. 4. Mobilize the nerve gently using fine watchmaker forceps, then lift it and place a prolene 7/0 ligature through the midpoint of the nerve just cranial to the branch running to the musculus biceps femoris (see Fig. 1A; Note 1). This procedure is to prevent the transection of more than half of the diameter of the nerve (see Note 2).

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