Gonadoctomy in Rat/ Mice

Instruments:
Trade names are occasionally used for identification purposes only and do not imply
endorsement.

1. Sterile instruments (scissors, tooth, and blunt forceps).

2. Metal clips and applying forceps.
3. Swab.
4. Sterile syringes (1 and 5 mL).
5. Hypodermic needles (25 gauge).
6. Infrared lamp.
7. Electric clipper.
8. Beaker.

Anesthesia :
1. Isoflurane.
2. Ketamine hydrochloride (Vetalar V™) and medetomidine hydrochloride
(Dormitor™) cocktail or Ketamine hydrochlo- ride (Vetalar V™) and Xylazine
(Rompun™) cocktail.
3. Atipamezole hydrochloride (Antisedan™).
Methods:

Animal Husbandary :

1. House animals in pathogen-free rooms maintained at constant temperature, with

12-h light/12-h dark cycles.
2. Provide free access to water and standard, pelleted commercial diet.
3. Allow animals an adaptation/acclimatisation period of at least 3 days prior operating if
transported from different facility.
Anaesthesia :

1. Induce anaesthesia by intraperitoneal injection of a cocktail of Vetalar V™ (females, 75

mg/kg; males, 50 mg/kg) in combi- nation with Dormitor™ (females, 1 mg/kg; males,
0.5 mg/kg) (see Note 1).
2. Check the depth of anaesthesia by monitoring respiratory rate (anaesthetised animals show
reduced respiratory rate) or sim- ply testing the animal response to gentle pressure on the
hind paws.

Preoperative Care :

1. After the onset of anaesthesia, place animals under infrared lamp to prevent heat loss.
2. Using an electric clipper, shave fur bilaterally over the lumbar spine (Ovx) or ventral side
of the scrotum (Orx) to expose skin (see Note 2).
3. Swab the shaved skin with 70% (v/v) ethanol followed by ster- ile PBS.
4. Ensure that all experimental protocols are approved and con- ducted in accordance with
regulation provided by regulatory ethics committee.
5. Sterilise and disinfect all surgical instruments and hard surfaces with 70% ethanol prior to
use.

Operative Technique for Ovariectomy :

1. Place the anaesthetised animal on the operating table with its back exposed and its
tail towards you (Fig. 1a).
2. Make a single midline dorsal incision (0.5 cm for mice and 2 cm for rat)
penetrating the skin using small scissors (see Fig. 1a). Incision should be made in
the lower back, directly below the bottom of the rib cage (see Note 3).
3. Gently free subcutaneous connective tissue from the underly- ing muscle on each
side using blunt forceps (see Fig. 1b).
4. Locate ovary under the thin muscle layer and make a small incision (less than 1 cm)
on each side to gain entry to the peri- toneal cavity (see Fig. 1b).
5. Hold securely the edge of the incision with tooth forceps and retract the ovarian fat
pad surrounding ovaries with blunt for- ceps to expose oviduct (see Fig. 1c).
6. Identify and replace ovaries back into the abdominal cavity for sham operations.
7. Perform a single ligature around the oviduct (0.5 cm for mice and 2 cm for rats from
ovary) to prevent bleeding following removal of ovary (see Fig. 1d).
8. Remove ovary by gently severing the oviduct, using sterile, small scissors (see Fig.
 1e).
9. Replace uterus and remaining part of the oviduct back into the abdominal cavity.
10. Suture the muscle layer (see Note 4).
11. Turn the animal over so that it is still laid on its ventral surface but its tail is pointing
away from you, and repeat the procedure described in steps 3–10 for the other ovary
if using one skin incision (see Fig. 1f and Note 2).
12. Close the skin incision using metal clips (see Note 5).

Fig. 1. Illustration of ovariectomy in the mouse. The ovary is indicated by the arrow.

Operative Technique for Orchidectomy :

1. Place the anaesthetised animal on the operating table on its Technique back with the tail
towards you for Orchidectomy.
2. Make a single incision on the ventral side of the scrotum (0.5 cm for mice and 1.5 cm for
rat) penetrating the skin using sterile scalpel (see Fig. 2a, b).
3. Localise the testicular fat pad on the left side and pull it through the incision using blunt
forceps (see Fig. 2c).
4. Cut the cremaster muscles and locate the testicular fat pad and gently pull it through the
incision using sterile, blunt forceps.
5. Expose testicular content by gently freeing testicular fat pad with sterile, blunt forceps
(see Fig. 2d).
6. Gently expose the cauda epididymis, caput epididymis, vas def- erens, and testicular
blood vessels while holding the testicular sack with sterile tooth forceps.
7. Perform a single ligature around the blood vessels to prevent bleeding following removal
of testis.
8. Gently severe cauda epididymis and caput epididymis from the testis (see Fig. 2e).
9. Remove testis by gently severing blood vessels with small scis- sors (see Fig. 2f).
 10. Replace the remaining content of testicular sac back using blunt forceps.
11. Repeat steps 1–9 for the other testis.
12. Close the skin with metal clips (see Note 5).

Fig. 2. Illustration of orchidectomy in the rat.

Post-operative Care
1. Reverse anaesthesia using an intraperitoneal injection of atipa- mezole hydrochloride
(Antisedan, 1 mg/kg for both males and females).
2. House animals individually and keep under close observation for approximately 2–4
h until they fully recover from anaesthesia.
3. Following recovery period (approximately 24 h after surgery), the animals can be
grouped together as normal.
4. Delay administration of experimental treatments for at least 24 h after surgery.

Risks in Oophorectomy

An oophorectomy is a relatively safe procedure. However, with any surgical procedure, there are
risks involved.

Risks of an oophorectomy include the following:

 Bleeding
 Infection
 Damage to nearby organs
 Rupture of a tumor, spreading potentially cancerous cells
 Retention of ovary cells that continue to cause signs and symptoms, such as pelvic pain,
in premenopausal women (ovarian remnant syndrome)
 Inability to get pregnant on your own, if both ovaries are removed
Menopause after oophorectomy

If you haven't undergone menopause, you will experience menopause if both ovaries are
removed. This deprives the body of the hormones, such as estrogen and progesterone, produced
in the ovaries, leading to complications such as:

 Menopause signs and symptoms, such as hot flashes and vaginal dryness
 Depression or anxiety
 Heart disease
 Memory problems
 Decreased sex drive
 Osteoporosis
An oophorectomy can be performed two ways:

 Laparotomy. In this surgical approach, the surgeon makes one long incision in your
lower abdomen to access your ovaries. The surgeon separates each ovary from the blood
supply and tissue that surrounds it and removes the ovary.
 Minimally invasive laparoscopic surgery. In this surgical approach, the surgeon makes
a couple of very small incisions in your abdomen.
The surgeon inserts a tube with a tiny camera through one incision and special surgical tools
through the others. The camera transmits video to a monitor in the operating room that the
surgeon uses to guide the surgical tools.
Each ovary is separated from the blood supply and surrounding tissue and placed in a pouch.
The pouch is pulled out of your abdomen through one of the small incisions.

Complications and risks in Orchiectomy

As with any surgical procedure, orchiectomy does come with a risk of post-operative side
effects and complications.

Some potential side effects associated with orchiectomy include:

 loss of muscle mass

 hot flashes
 fatigue
 decreased sexual interest
  gynecomastia, or breast enlargement

Some potential complications associated with orchiectomy include:

 extensive bruising
 blood clots
 hemorrhaging, or internal bleeding
 infertility
 erectile dysfunction
 osteoporosis
 infection