Anesthetic: Isoflurane
An anesthetized rat is immobilized in a stereotaxic frame in flat skull position. A 2 cm midsagittal skin incision is made on the scalp to expose the skull. An infusion cannula consisting of a sterilized length of 30 gauge stainless steel hypodermic tubing is stereotaxically placed via a hole in the skull and advanced so that the internalized tip is located within the nigrostriatal pathway. The stereotaxic coordinates are determined a priori and verified by quantification of apomorphine induced rotational activity in sentinel animals of the same strain, sex and weight range as specified in the order. (For WH or SD male rats 175-225grams, the coordinates from Bregma are: AP-4.3, ML +1.2, DV -8.3) A 2 ug/ul solution of 6-hydroxydopamine is infused at a rate of 1 ul/min for a 4 minute duration into the nigrostriatal pathway. Following drug infusion, the infusion cannula is allowed to sit in place for 5 minutes, then slowly withdrawn and the skin incision closed with stainless steel wound clips.
Anesthetic: Isoflurane
A 2 cm dorsal midline skin incision is made with its cranial terminus at the level of the 13th rib. The muscle wall is opened - 2 cm lateral to the spine. The adrenal is located cranial and medial to the kidney and is imbedded in a fat pad. Using blunt forceps, the entire fat pad is drawn out of the incision, and the adrenal removed intact. No ligature is necessary. The skin incision is close with wound clips. All adrenalectomized animals are maintained and shipped with 0.9% sodium chloride solution, ad lib.
Note: Due to strain sensitivity, this procedure is not available in FVB mice.
The methodology for sham adrenalectomies is identical, with the exception that the adrenals are left intact.
Adrenalectomy Related Publications
Anesthetic: Isoflurane
A 2 cm dorsal midline skin incision is made with its cranial terminus at the level of the 13th rib. The muscle wall is opened 2 cm lateral to the spine. The adrenal is located cranial and medial to the kidney and is embedded in a fat pad. With blunt forceps, the entire fat pad is drawn out of the incision, the adrenal capsule is ruptured and the contents expressed. The remains of the capsule are replaced in the fat pad, and the fat pad is replaced. No ligature is necessary. The muscle incision is approximated, and the skin incision closed with wound clips. Animals are maintained and shipped with 0.9% sodium chloride solution for 48 hours post surgery.
The sham procedure is identical in that the adrenals are located but no rupturing of the capsule occurs.
Anesthetic: Isoflurane
A 2 cm ventral midline incision is made in the scrotum, and the skin retracted to expose the tunica. The tunica is pierced, and the opening stretched with blunt forceps. The testis is pushed out by gentle pressure on the pelvic region. The spermatic artery is clamped and cauterized, and the testis removed. The epididymus, deferential vessels and ductus are replaced in the tunica. Wound clips close the incision.
The methodology for sham castration is identical, differing only in that the testis is replaced in the
tunica intact.
Anesthetic: Xylazine/Katamine (IP)
A ventral midline incision of 3 cm length is made with its cranial terminus just caudal to the xiphoid process. The incision is opened with retractors and the liver located. The median and left lateral lobes are removed. One lobe at a time is isolated, and the distal portion removed by ligating the blood vessels leading to the organ. The muscle incision is closed with sutures and the skin incision is closed with wound clips.
Sham hepatectomies are performed identically with the exception that the lobes are only located.
Anesthetic: Sodium Pentobarbital (IP)
Animal is mounted on the hypophysectomy instrument. Fill syringe, which is fitted with stop equipped needle with roughly 2 cc of sterile saline. Insert the needle with the screw through the hollow right ear bar until resistance is met. Rotate the needle clockwise 3600.Keeping the same needle orientation, fully insert needle to the stop. Rotate needle to cut the pituitary stalk. Remove the pituitary by gentle aspiration – gently withdraw the plunger until the pituitary gland enters the glass syringe. Discharge the aspirate from the syringe into a sterile petri dish. Inspect the gland for completeness of removal. Animals are maintained and shipped a with 5% sucrose solution, ad lib.
Sham hypophysectomies are performed in the identical manner, with the exception that the needle is inserted to the point of initial resistance. No cutting or gland aspiration takes place.
Hypophysectomy Related Publications
Anesthetic: Isoflurane
An anesthetized rat is place in a stereotaxic hypophysectomy board, and the snout is brought as close to the ventral surface of the body as possible. A 2 cm skin incision is made caudal to the base of the skull, over the area where the spine joins to the skull. The underlying muscles are cleared with blunt dissection. At the white area, which indicates the cranial end of the spinal column, a 25 G needle is used to form a small hole. The cannula is inserted through this hole and tied above and below the silicone bubble with 3/0 silk ligatures. The skin incision is closed with stainless steel wound clips.
A sham intrathecal cannulation can be performed identically, with no insertion of the cannula taking place.
Anesthetic: Isoflurane
A microdialysis guide cannula is implanted at specific target areas of the brain. Target areas include: hippocampus, prefrontal and striatal portions of the brain as well as custom coordinates available upon request. In rats, 10mm stock MD guides (please specify MD-2250 or 2251) are inserted to the appropriate depth and secured by three stainless steel trephine screws and cranioplastic cement.
Anesthetic: Isoflurane
A 2 cm dorsal midline skin incision is made with its cranial terminus at the level of the 13th rib. The abdominal wall is opened 1.5 to 2 cm lateral to the spine. With blunt forceps, the kidney is seized and retracted through each incision. All blood vessels and the Ureter are ligated and the organ removed. The muscle is approximated and the skin incision closed with wound clips.
Sham nephrectomies are performed identically in all respects, with the exception that no ligation takes place and the kidney is replaced intact.
Anesthetic: Isoflurane
A 2.0 cm skin incision is made in the ventral midline, with its cranial terminus 1.0 cm caudal to the xyphoid process. A 2.0cm muscle incision is made along the midline. The right kidney is isolated and cleared of surrounding fat and connective tissue to clearly view the renal artery and vein, and ureter as they enter the hilus of the kidney. Care is exercised to minimize disturbance of the adrenal gland. A ligature (3/0 silk) is placed around the renal artery, vein and ureter. These vessels are then cut proximal to the kidney and the kidney is removed, taking care that the adrenal gland is not disturbed. The left kidney is then isolated and cleared of surrounding fat and connective tissue to clearly view the renal artery and vein. Using the dissecting microscope, the three branches of the renal artery proximal to the kidney are identified. Fine ligature (5-0 silk) is used to ligate two of the branches. As these ligatures are tightened, the kidney takes on an ischemic coloration which should encompass about 2/3 of the kidney surface area.
The muscle incision is closed with silk sutures. The skin incision is closed with wound clips.
Post-operative Analgesics: Not required
Reference: Wada,M et al; The Calcimimetic Compound NPS R-568 Suppresses Parathyroid Cell Proliferation in Rats with Renal Insufficiency; J Clin Invest 1997 Dec 15;100(12):2977-83
Restrictions: This procedure has not been validated in juvenile animals as of 3/01/99. This procedure is only available in adult rats, bodyweight of 200 grams or greater.
Anesthetic: Isoflurane
A 1.5 cm dorsal midline incision is made with its cranial terminus 1.5 to 2.5 cm caudal to the 13th rib, and any fascia trimmed away. The muscle wall is pierced with forceps, 1.5 to 2 cm lateral to the spine. The ovary is located in a fat pad under the dorsal muscle mass. With blunt forceps, it is drawn through the muscle and skin incisions, clamped and ligated to the level of the fallopian tube. The ovary is removed and the uterine horn returned to the body cavity. The muscle is approximated and the skin incision closed with wound clips. The other ovary is removed similarly.
A sham ovariectomy is performed identically, but the ovaries remain intact.
Ovariectomy Related Publications
Anesthetic: Isoflurane
A ventral midline incision of 2 cm is made, its caudal terminus at the level of the clavicle. The underlying salivary and lymphatic tissues are pushed cranially and the omohyoideus muscle divided longitudinally. Retractors are applied to yield a clear view of the larynx and thyroids. The parathyroids are seen as minute white areas on the cranial surfaces of the glands. A cautery tip is applied to the parathyroids and nearby thyroid tissue. The omohyoideus is approximated, salivary and lymphatic tissues replaced, and the skin incision closed with wound clips. All parathyroidectomized animals are maintained and shipped with 2% calcium lactate solution, ad lib.
Sham parathyroidectomies are performed in an identical manner, with the exception that the cautery tip remains unheated.
Anesthetic: Isoflurane
A 2.5 cm midline ventral skin and abdominal muscle wall incision is made in the lower abdominal cavity. The cecum is gently exteriorized and placed on sterile draping material. Place draping material around the incision. A branch of the mesenteric venous system, the ileocolic vein, is visualized running parallel to the rostrocaudal axis of the rat. A cannula consisting of sterilized silicon rubber tubing with a polyurethane tippet is inserted into the lumen of the ileocolic vein via venotomy. The tip is advanced into the portal vein to the vicinity of the junction with the lineal vein and secured. The viscera is repositioned into the abdominal cavity and the abdominal wall incision closed with silk suture material. The cannula is subcutaneously tunneled to exit a small skin incision over the scapulae. The ventral abdominal skin incision is closed with wound clips. Cannula patency is verified by blood sample withdrawal. The cannula is filled with a viscous heparin solution to preclude thrombus occlusion and the sampling/infusion port closed with a short length of 23 gauge (.025" OD) stainless steel wire. The externalized terminus of the cannula is secured dorsally by passing it through a wound clip used to close the dorsal skin incision. Patency is again verified on the day of shipment by means of blood sample withdrawal.
Anesthetic: Isoflurane
A 2 cm dorsal longitudinal skin incision is made slightly to the left of midline over the pelvic girdle. Slightly caudal and ventral to the left hip joint, a faint separation of muscle groups is visualized. A 10 mm incision is made along the separation of the muscle groups and deepened by blunt dissection. The sciatic nerve is visualized running parallel to the length of the femur. Under 7 x magnification; a 7-10 mm length of the sciatic nerve, proximal to the sciatic trifurcation is carefully cleared from underlying tissue using blunt dissection. The nerve crush is performed using hemostatic forceps, situated perpendicular to the sciatic nerve, by closure of the hemostatic forceps to the first click for a 30 second duration. After removal of hemostatic forceps, this injury should result in a visible compression of the entire cross sectional area of the sciatic nerve. The muscle groups approximated and skin incision is closed with wound clips.
The sham procedure is performed in an identical manner, with the exception that no crushing of the sciatic nerve takes place.
Anesthetic: Isoflurane
A 2 cm dorsal longitudinal skin incision is made slightly to the left of midline over the pelvic girdle. Slightly caudal and ventral to the left hip joint, a faint separation of muscle groups is visualized. A 10 mm incision is made along the separation of the muscle groups and deepened by blunt dissection. The sciatic nerve is visualized running parallel to the length of the femur. A 7-10 mm length of the sciatic nerve, proximal to the sciatic trifurcation is carefully cleared from underlying tissue using blunt dissection. The nerve is sectioned using fine (Vannas) scissors. The muscle groups are approximated, and the skin incision is closed with wound clips.
The sham procedure is performed on the right side in an identical manner, with the exception of sectioning the sciatic nerve.
Anesthetic: Isoflurane
A 2 cm dorsal midline skin incision is made with its caudal terminus at the level of the 13th rib. The spleen is exposed by opening the abdominal wall on the anatomical left, 1.5 to 2.5 cm from midline. With blunt forceps, the organ, (with accompanying blood vessels and pancreatic tissue), is exteriorized through the incision. The blood vessels are ligated and the spleen removed. The pancreas is replaced, muscle approximated, and the skin incision closed with wound clips.
Sham splenectomies are performed identically, with the exception that no ligation takes place and the spleen is replaced intact.
Anesthetic: Isoflurane
A ventral midline skin incision 2 cm long, with its caudal terminus at the level of the clavicle, is made. Underlying lymphatic and salivary tissues are pushed cranially, and the omohyoideus and sternohyoideus muscles retracted to yield a clear view of the carotid artery. The carotid is followed cranially and the superior cervical ganglion located adjacent to the bifurcation of the carotid. The ganglion is excised using fine scissors. The lymphatic and salivary tissue is approximated and the skin incision closed with wound clips.
The sham procedure is performed in an identical manner, with the exception that no excision of the ganglion takes place.
Anesthetic: Xylazine/Ketamine (IP)
A ventral midline incision of 2 cm is made, its caudal terminus at the level of the clavicle. The underlying salivary and lymphatic tissues are pushed cranially and the omohyoideus muscle divided longitudinally. Retractors are used to give a clear view of the larynx and thyroid. The thyroid is removed by dividing the isthmus and gently teasing the glands loose. Care must be taken not to disturb the recurrent laryngeal nerve. Any bleeding is controlled with sterile cotton swabs. The thyroid glands are placed in a petri dish where the parathyroids are gently dissected away and placed back into the thoracic cavity alongside the trachea. The muscle is approximated, salivary and lymphatic tissue replaced, and the skin incision closed with wound clips. All thyroidectomized animals are maintained and shipped with 2% calcium lactate solution, ad lib.
Sham thyroidectomies are performed identically, but the thyroid is not disturbed.
Anesthetic: Xylazine/Ketamine (IP)
A ventral midline incision of 2 cm is made, its caudal terminus at the level of the clavicle. The underlying salivary and lymphatic tissues are pushed cranially and the omohyoideus muscle divided longitudinally. Retractors are used to give a clear view of the larynx and thyroid. The thyroid is removed by dividing the isthmus and gently teasing the glands loose. Care must be taken not to disturb the recurrent laryngeal nerve. Any bleeding is controlled with sterile cotton swabs. The muscle is approximated, salivary and lymphatic tissue replaced, and the skin incision closed with wound clips. All thyroidectomized animals are maintained and shipped with 2% calcium lactate solution, ad lib.
Sham thyroidectomies are performed identically, but the thyroid is not disturbed.
Anesthetic: Isoflurane
A 1 cm ventral midline incision is made, with its caudal end near the penis. Looking caudally through the incision, the massive vas deferens are immediately visible. Care must be taken not to cut the deferential blood vessels while sectioning ducts. A 0.5 cm section of each duct is removed by use of a cautery knife. The abdominal wall is closed with silk suture and the skin incision is closed with wound clips.
The sham vasectomy is performed identically without the ducts being sectioned.
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