Urine Depletion and Bladder Rinsing in Mouse Model: A Catheter-based Procedure to Empty and Wash Murine Urinary Bladder

Published: April 30, 2023

Abstract

Source: Yu, C. et al. Novel Method of Plasmid DNA Delivery to Mouse Bladder Urothelium by Electroporation. J. Vis. Exp. (2018)

In this video, we describe a catheter-based procedure to empty and rinse the urinary bladder of a mouse model. The prepped mouse can then be used for further experimental procedures.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

 1. Anesthetizing Animal

  1. Use a tabletop anesthesia system to anesthetize the animal with isoflurane. Turn on the O2 tank and adjust the oxygen flowmeter to 1 L/min.
  2. Place an adult female C57BL/6J mouse in the induction chamber. Turn on and adjust the isoflurane vaporizer to 3% for induction. The mouse should be in anesthesia within 2 min. Administer buprenorphine analgesic (0.1 mg/kg) subcutaneously upon removal of the mouse from the induction chamber for preemptive analgesia.
    NOTE: In this protocol, female mice are used, as they are easier to work with during urethra catheterization. The protocol also works for male mice, but extra caution is needed.
  3. Apply ophthalmic ointment to the eyes of the animal to prevent dryness. Place the anesthetized mouse facing up on a heating pad with its nose in the nose cone. Turn the air circuit switch to let isoflurane go through the nose cone.
  4. Restrain the head and the nose cone with adhesive tape. Adjust the isoflurane vaporizer to 2% for maintenance.
  5. Restrain the limbs with adhesive tape. Perform toe-pinch tests to make sure the animal is in deep anesthesia and then shave the abdominal area.

2. Urine Depletion and Bladder Rinsing

  1. Apply lubricant to the catheter (24G, length of 2.11 cm, outer diameter of 0.045 cm) and ensure that its outer surface is fully covered.
  2. Insert the catheter into the urethra opening and slowly push it until it reaches the bladder, at which time the urine should flow out through the catheter. Gently press the abdomen to help urine depletion.
    1. Check if the catheter is inserted correctly into the bladder by observing automatic urine out-flow. Avoid piercing the urethra and bladder wall and apply lubricant multiple times if needed.
  3. Discard the urine with a pipette into a waste beaker.
  4. Pipette 80 µL of phosphate-buffered saline (PBS) into the outer end of the catheter, with the other end still in the bladder. Carefully attach a 1 mL syringe to the catheter. Gently push the syringe to inject the PBS into the bladder. Leave some PBS in the catheter to avoid creating air bubbles in the bladder.
  5. Remove the syringe. Wait for PBS to drain out of the bladder. Gently press the abdomen to evacuate PBS.
  6. Discard the PBS in the catheter with a pipette into the waste beaker.
  7. Repeat PBS washing (steps 2.4 to 2.6) two more times.
    NOTE: It is essential to work gently during these washing steps. Blood in the catheter or failure of automatic out-flow of liquid usually indicates the urethra is pierced through

Divulgaciones

The authors have nothing to disclose.

Materials

BD 1mL TB Syringe Fisher 148232E
Exel International Disposable Safelet I.V. Catheters Fisher Scientific 14-841-21 24G, 2.11 cm length, 0.045 cm outer diameter
K&H Heated Resting Mat for Small Animals, 9 By 12 Inches Amazon Cover the heat mat with paper towels
Ophthalmic ointment Fisher NC0849514
PBS, pH7.4 Life Technologies 10010049
Pipet tips 200 µL USA Scientific 1111-0206
Ophthalmic ointment  Fisher  NC0849514
Pipet Tips, Universal Fit; 0.1 to 10 µL Fisher 02707438(CS)
Buprenorphine  Sigma  B9275-50MG
PIPETMAN NEO P2 Gilson F144561
PIPETMAN NEO P200N Gilson F144565
Therio-gel Veterinary Lubricant PBS animal health 353-736
V-1 Table top system anesthesia VetEquip 901806
Isoflurane  Vet one  501017
Tape, Scotch Fisher 19047257

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Urine Depletion and Bladder Rinsing in Mouse Model: A Catheter-based Procedure to Empty and Wash Murine Urinary Bladder. J. Vis. Exp. (Pending Publication), e20563, doi: (2023).

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