Rodent Stereotaxic Surgery and Animal Welfare Outcome Improvements for Behavioral Neuroscience

Summary

Stereotaxic surgery on rodents allows for targeted administration of drugs or electrical stimulation and recordings in awake, behaving animals. In this video presentation we will demonstrate recent procedural refinements to this long-standing procedure that successfully improved survival rate and reduced post-surgical weight loss.

Abstract

Stereotaxic surgery for the implantation of cannulae into specific brain regions has for many decades been a very successful experimental technique to investigate the effects of locally manipulated neurotransmitter and signaling pathways in awake, behaving animals. Moreover, the stereotaxic implantation of electrodes for electrophysiological stimulation and recording studies has been instrumental to our current understanding of neuroplasticity and brain networks in behaving animals. Ever-increasing knowledge about optimizing surgical techniques in rodents^1-4, public awareness concerning animal welfare issues and stringent legislation (e.g., the 2010 European Union Directive on the use of laboratory animals⁵) prompted us to refine these surgical procedures, particularly with respect to implementing new procedures for oxygen supplementation and the continuous monitoring of blood oxygenation and heart rate levels during the surgery as well as introducing a standardized protocol for post-surgical care. Our observations indicate that these modifications resulted in an increased survival rate and an improvement in the general condition of the animals after surgery (e.g. less weight loss and a more active animal). This video presentation will show the general procedures involved in this type of stereotaxic surgery with special attention to our several modifications. We will illustrate these surgical procedures in rats, but it is also possible to perform this type of surgery in mice or other small laboratory animals by using special adaptors for the stereotaxic apparatus⁶.

Protocol

Note: Antiseptic techniques should be employed during the whole procedure. All the instruments and materials (cotton-tipped swabs, gauze, etc.) that will be used during the surgery should be sterilized by autoclaving. A surgical mask, hair bonnet and sterile gloves should be worn. The working area and the stereotaxic apparatus should be cleaned thoroughly, and disinfected with a 70% ethanol solution.

1. Pre-surgical procedures

1. Set up the stereotaxic apparatus and all materials needed. Pre-warm the heating pad.
2. Place the cannula in its support and check if it is straight.
3. Turn on the gas system - mixture of am....

Discussion

The main purpose of this video presentation is to familiarize behavioral neuroscientists with the basic principles of stereotaxic surgery. Researchers who are already performing stereotaxic surgery might also benefit from this video and consider some of the procedural refinements for use in their own laboratory. An ever-increasing knowledge about optimizing surgical techniques^1-3, the development of new anesthetics and analgesics for use in human and veterinary medicine^4,12, public awareness concern.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Alcohol 70%	VWR international
Antisedan (atipamezole)	Orion
Atropine sulfate 0,5	Teva Pharmachemie
Bulldog haemostatic clamp	Aesculap
Cannulas	Component Supply Co.
Chlorhexidine 0.5%	APP Pharmaceuticals
Cleaning powder	Alconox, Inc.
Cotton sticks	Hartmann BV
Dexdomitor (dexmedetomidine)	Orion
Drill	Dremel
Duratears Z	Alcon
Durogrip Naaldv–rder converse 130mm	Aesculap
Durotip Fijne schaar ret.sp/st 110 mm	Aesculap
Gauze	Medicomp (5x5)
Heating pad	Harvard Apparatus
Insect pins (stylets)	Entosphinx (Czech Republik)
Ketamine 10% (ketamine)	Alfasan
Micro-halsted straight tip	Aesculap
Molinea	Hartmann BV
NaCl 0,9%	Baxter Internationl Inc.
Needles (25G)	Terumo Medical Corp.
Oximeter	Edan Instruments, Inc.
Pentobarbital	Pharmacy of the UMCG
Rimadyl	Pfizer Pharma GmbH
Scalpel blade No. 23	Swann-Morton
Scalpelholder NR. 4 133 mm	Aesculap
Screw driver	Any Supplier
Simplex Rapid (dental cement)	Kemdent
Skull screws	Component Supply Co.
Spatula	VWR international
Spongestan special	Johnson & Johnson
Stereotacts	Kopf Instruments
Surgical forceps 100mm	Aesculap
Suture material Safil 5/0	Aesculap
Syringe 10 ml	Omnifix
Syringe 1ml	Terumo Medical Corp.
Syringe 5ml	Omnifix
Xylocaine (lidocaine/adrenaline)	AstraZeneca