performing electroacupuncture on rat immobilized using rat jacket

Rat Jackets to Enhance Suspension Fixation During Electroacupuncture on Rats

Acupuncture is a traditional therapy characterized by needling at specific acupoints or other positions to prevent and treat diseases. Electroacupuncture combines filiform needles with electrical stimulation1, which allows for precise control over the amount of stimulation by adjusting relevant parameters, promoting the standardization of acupuncture treatment2. In recent years, research on the mechanisms of electroacupuncture has continued to expand and deepen3,4, which has led to an annual increase in studies focusing on the mechanisms of electroacupuncture5.
In basic research on electroacupuncture, rats are the most commonly used animals. During electroacupuncture treatment, the recipient must remain still to ensure a smooth procedure. Unlike in clinical trials with human subjects, small animals used in animal experiments cannot follow commands consciously during 15-30 min lengthy electroacupuncture procedures. This presents researchers with a primary challenge of achieving stable and reliable fixation. Improper fixation can lead to negative emotions such as anxiety, depression, and fear in animals6, which not only increases the risk of injury for rats and the chance of being bitten for the experimental personnel but also affects the stability and persuasiveness of research results.
In previous studies, researchers have often used anesthesia fixation7, high platform fixation8, binding fixation6,9, or rat coat fixation10 before acupuncture or electroacupuncture operations. However, given the effects of anesthesia on the body11 and brain function12, and considering that acupuncture is performed on conscious patients in clinical cases, anesthesia may impact research results to a certain extent. Therefore, compared with anesthesia fixation, comfortable fixation of rats in an awake state is a better option.
However, in the non-anesthetized state, various fixation methods also possess their respective limitations, including restricted acupoint exposure due to coverage of the rat coat on rats10, the necessity of at least two individuals to perform the operation13, non-adjustable fixator size14, or the stressed state of rats in a forced position6. To address these technical issues, we designed a rat jacket and a fixation rack suitable for acupuncture manipulation in most parts of the rat body. This can help researchers efficiently carry out acupuncture procedures with rats.
The rationale for this protocol is as follows: firstly, the closed headgear on the rat&#39;s jacket takes advantage of the rats&#39; natural inclination for darkness and burrowing, thus restricting their range of activities. Secondly, the binding ropes near the joints of the rats&#39; limbs provide physical compression restraint, imposing certain limitations on their movements. However, rats are still able to jump and move horizontally even while wearing the jacket. Lastly, since rats cannot touch the ground without a fixed force point when suspended, we added a suspension fixation device to the rat jacket, further restricting their movements and ensuring needle retention. The composition of the fixation device is shown in Figure 1.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/65652/65652fig01.jpg" /> 
Figure 1: Schematic diagram of the rat jacket and suspension fixation. (A) The general appearance of the rat jacket in a flat view. (B) Three-dimensional view of the rat jacket. (C) Specially tailored rat jacket with an incision near the acupoints on the head and back waist. (D) Illustration of multiple rats simultaneously suspended on the elevated rack. 1: Fabric overall view; 2: Forelimb outlet; 3: Hind limb outlet; 4: Tying rope; 5: Cavity formed after suturing; 6: Incision for exposing head acupoints; 7: Incision for exposing back and waist acupoints. 8: Suture position. <a href="https://www.jove.com/files/ftp_upload/65652/65652fig01large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>

Author Spotlight: Efficient Rat Clothing for Improved Rat Fixation During Electroacupuncture 

Improved Suspension Fixation Using Rat Jacket in Rat Acupuncture Experiments

We developed an improved method for suspending and fixing rats with rat clothes in an acupuncture experiment. This protocol aims to provide detailed descriptions of its procedural steps, including important precautions, offering a reference for researchers conducting fundamental experiments in the field of acupuncture and moxibustion. In previously reported studies, researchers have often used anesthesia fixation, high platform fixation, binding fixation, or various other self-made fixators while performing acupuncture or electroacupuncture operations on rats.Given the side effects of anesthesia on the body and the real clinical conditions, fixation in an awakened stage offers the better alternative. However, in the non-anesthetized stage, the mentioned fixations also possess limitations such as restricted acupoint exposure, requirement of multiple labels, non-adjustable fixator size, or forced posture. This method utilizes the rat's preference for burrowing to create rat jackets that fit their body shape.It offers several advantages over other techniques. It is cost-effective, user-friendly, manpower-and space-saving, accommodates rats of varying weights, and can also be used for needle operation on most acupoints. In subsequent studies, we will focus on investigating the impact of the restraints and suspensions of rat clothing on physiological indicators and behavioral aspects related to emotional cognition in rats.This will enable us to quantitatively assess its potential influence on research and refine the fixation method.

Watch this Scientific Journal Video about Performing Electroacupuncture on Rat Immobilized Using Rat Jacket at JoVE.com

Performing Electroacupuncture on Rat Immobilized Using Rat Jacket

Before beginning the procedure of electropuncture on the rat, dress the rat in the custom made rat jacket. Then disinfect the local skin of acupoints using alcohol swabs. After locating the acupoint accurately, insert the sterile disposable acupuncture needle into the spot at the appropriate depth.Gently suspend the animal from the horizontal iron rod with the help of the S-shaped hooks attached to the rat jacket. Wrap the electrode wire around the horizontal iron rod before securely attaching the small clip at the end of the electrode wire to the needle handle. Then after switching on the power, select the appropriate frequency, current, and time.Once the parameters are adjusted, commence the electroacupuncture treatment process. Using the S-shaped hooks, suspend the rats in the same group together on the iron rod for simultaneous electroacupuncture. When the treatment is over, remove the acupuncture needle along with the clamp from the animal.Then carefully take the needle out of the clamp and put it in the box. After removing the animal from the suspension hooks, loosen the straps, lift the head cover of the rat's jacket and position the rat above the cage. The rat will then be free to release itself from the jacket and enter the cage independently.While performing a 15 minute electroacupuncture experiment on 12 rats only one rat exhibited intense struggle during the needle retention period resulting in needle drop. While other rats maintained proper needle fixation throughout the treatment. When compared to the other fixation methods reported in the literature, the demonstrated method exhibited the combined benefits of stability and secure fixation, efficiency in terms of time and number of personnel required, affordability, as well as relative comfort for the rats.Additionally, in terms of the exposed acupoint range, the specially designed rat jackets in this protocol met the requirements for exposing most acupoints on the body.

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Research

JoVE Journal

Medicine

163 vistas.  Hubei University of Chinese Medicine. Antes de comenzar el procedimiento de electropuntura en la rata, vítala con la chaqueta de rata hecha a medida. A continuación, desinfecte la piel local de los puntos de acupuntura con hisopos con alcohol. Después de ubicar el punto de acupuntura con precisión, inserte la aguja de acupuntura desechable estéril en el lugar a la profundidad adecuada.Suspende suavemente al animal de la barra de hierro horizontal con la ayuda de los ganchos en forma de S unidos a la chaqueta de rata. ...