securing the rat in the rat jacket for electroacupuncture

מעילי חולדות לשיפור קיבוע המתלים במהלך דיקור חשמלי בחולדות

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>

מחבר זרקור: בגדי חולדות יעילים לשיפור קיבוע חולדות במהלך דיקור חשמלי 

קיבוע מתלה משופר באמצעות מעיל חולדה בניסויי דיקור חולדות

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 Securing the Rat in the Rat Jacket for Electroacupuncture at JoVE.com

אבטחת החולדה במעיל החולדה לדיקור חשמלי

לבשו זוג כפפות לפני שתמשיכו להתאים ולאבטח את החולדה במעיל עכברוש בהתאמה אישית. לאחר מכן, ביד אחת, החזיקו את החלק האמצעי והאחורי של מעיל החולדה על השולחן. בעזרת היד השנייה, החזיקו בעדינות את החולדה בזנבה כדי להרים אותה מכלוב ההאכלה והכניסו אותה לתוך הז'קט כשהחולדה נעה קדימה לתוך הז'קט וראשה מגיע לנקודת כיסוי הראש.בעזרת שתי ידיו, משוך בזהירות את גפיו העליונות השמאלית והימנית מהחורים בז'קט. כרכו היטב את חבלי הקשירה מקצה הפתיחה של הז'קט סביב תא המטען של החולדה, וקשרו אותם בחוזקה בקצה הנגדי כדי לרסן כראוי את גפיה העליונות. קשרו את החבל בצורה רופפת מספיק כדי למנוע היפוקסיה או איסכמיה מקומית של הגפיים, והדוקים מספיק כדי למנוע את בריחת החולדה.באופן דומה, אבטח את גפיו התחתונות.

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Research

JoVE Journal

Medicine

Securing the Rat in the Rat Jacket for Electroacupuncture

176 Views.  Hubei University of Chinese Medicine. לבשו זוג כפפות לפני שתמשיכו להתאים ולאבטח את החולדה במעיל עכברוש בהתאמה אישית. לאחר מכן, ביד אחת, החזיקו את החלק האמצעי והאחורי של מעיל החולדה על השולחן. בעזרת היד השנייה, החזיקו בעדינות את החולדה בזנבה כדי להרים אותה מכלוב ההאכלה והכניסו אותה לתוך הז'קט כשהחולדה נעה קדימה ל...

Video: אבטחת החולדה במעיל החולדה לדיקור חשמלי

Improved Suspension Fixation Using Rat Jacket in Rat Acupuncture Experiments

In basic experimental acupuncture research, rats are commonly used as laboratory animals. However, it is difficult for them to maintain a fixed posture. During electroacupuncture procedures, proper immobilization of rats is essential. Various methods of rat fixation are currently used, including anesthesia fixation, high-platform fixation, binding fixation, and fixation with a self-made rat coat. However, these methods have their limitations, which may affect the efficiency and operability of the experiment to some extent. This protocol introduces a method of suspending and fixing rats using rat clothes. Firstly, rats are clothed with rat jackets that match their body shape, taking advantage of their preference for darkness and burrowing. The needling operation can then be carried out after the rats have worn rat clothes. When suspended, the rats are relatively still, as their limbs cannot move. This fixation method offers not only economical and user-friendly benefits but also ensures a stable and reliable fixation of the rats in a comfortably relaxed position. It also effectively minimizes time consumption, experimental space, and manpower resources. Additionally, this method allows for the exposure of most acupoints used for acupuncture in rats. This article primarily concentrates on the device's composition, encompassing a specially designed rat jacket, an elevated fixation rack, and their connecting structures. Additionally, an illustrative example will be presented to demonstrate the application of the rat clothing-based suspension fixation method in rat electroacupuncture procedures.

Here, we present an improved suspension fixation method using a rat jacket to effectively immobilize rats during acupuncture experiments. Furthermore, the structure and application of this fixation apparatus, as well as instructions and evaluation for its utilization in electroacupuncture therapy, is elucidated.

In basic experimental acupuncture research, rats are commonly used as laboratory animals. However, it is difficult for them to maintain a fixed posture. ...

Wear a pair of gloves before proceeding to fit and secure the rat in the custom made rat jacket. Then using one hand hold the rat jacket's middle and back portion on the table. Using the other hand, gently hold the rat by its tail to pick it up from the feeding cage and place it into the jacket as the rat moves forward into the jacket and its head reaches the headgear point.Using both hands, carefully pull out its left and right upper limbs from the holes in the jacket. Securely wrap the tying ropes from the opening end of the jacket around the rat's trunk, and tie them tightly at the opposite end to adequately restrain its upper limbs. Tie the rope loose enough to prevent hypoxia or local limb ischemia, and tight enough to prevent the rat's escape.In a similar manner, secure its lower limbs.

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.