Technique improvement and experience sharing of cannulation of the mouse submandibular salivary gland via the Wharton’s duct

Abstract

Objective This paper described my technique improvement and experience sharing of Cannulation of the mouse submandibular salivary gland via the Wharton’s duct in order to accelerate the submandibular gland gene therapy research in China. Methods The operation process of the technology was introduced in detail, and the improvement of the original technology was made. The micro pump and microsurgical instruments were introduced to make the operation more precise and precise, which made the success rate higher. Results On the basis of a certain surgical operation experience, we trained three times per week, every time three to five mouse. Due to inappropriate operation technique and unclear understanding of the anatomy, initially intubation success rate was very low. The location of intubation always was incorrect, intubation site bleeding aometimes happened, poor control of anesthesia led to the death of mice. With the accumulation of operating experience, intubation success rate rapidly improveed about four weeks later, even achieve 100% success rate. Different drugs need different injection speed. If we injection of saline only, there were no significant damage between injection speed 10 μl/min and 25 μl/min, the volume of 25 μl and 50 μl. If we injected nanoparticle+ small interfering RNA (siRNA, specific target gene siRNA), the injection speed of 25 μl/min can cause more damage than that of 10 μl/min.25 μl/min resulted in heavier tissue damage in female mice. Conclusion Retrograde intubation through the mouse Wharton’s Duct required some training to master. Familiar with the anatomy was the key. If the anatomical location was accurate, we could gradually training intubation techniques, force and the angle of the cannulation to continuously improve the success rate of this operation. Key words: Salivary glands; Submandibular gland; Gene therapy; Cannulation; Technology