Protocol
Jugular vein catheterization for repeated blood sampling in the unrestrained conscious rat

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Abstract

The ability to obtain repeated, low-stress blood samples from adult rats enables the design of complex experiments in which time course information or evaluation of repeated treatments is necessary. Furthermore, it reduces the number of animals necessary to acquire such information and, thus, facilitates compliance with the animal use 3Rs (reduction, refinement and replacement). To this end, a microsurgical technique to collect blood samples from the right atrium through a catheter (cannula) implanted into the right external jugular vein of adult rats is described. Rats tolerate this simple and efficient vascular access technique as evidenced by the absence of overt morbidity or abnormal behaviors. Blood is easily sampled while the rats reside in their home cages. Because the sample volume is replaced, repeated sampling is possible without compromising blood volume. Successful adoption of this procedure by other investigators will be aided by the photographic illustrations accompanying this detailed description of the procedure. Application of this technique to monitor temporal changes in plasma stress hormones during stressor paradigms as well as after behavioral and pharmacological challenges is discussed.

Section snippets

Type of research

Microsurgical technique for rat external jugular vein catheterization.

Time required

30 min, catheter preparation.

Overnight, catheter sterilization.

45–60 min, preoperative preparation: catheter, anesthetics, clean area, instruments, rat.

10–15 min (per animal) surgery for catheter implantation.

90–120 min postoperative recovery.

Surgical

An overview of the materials for the implantation of the jugular catheter is shown in Fig. 1. For the microsurgical technique, the specifications of the surgical instruments are more critical than their source. Accordingly, the name of a specific vender is not presented with each item. Items are obtained from one or more of the following sources: Biomedical Research Instruments (www.biomedinstr.com), Fine Science Tools (www.finescience.com), Fisher Scientific (www.fishersci.com), George Tiemann

Microsurgical technique

All procedures are approved by Emory University Institutional Animal Care and Use Committee (IACUC) and comply with NIH (http://grants.nih.gov/grants/olaw/olaw.htm) recommendations based on National Research Council guidelines [25]. For the illustration of the technique, an adult male Long Evans rat (300–325 g BW, Charles River, Portage, MI, USA) is used. However, the technique is applicable to rats of either sex, weighing 250 g and above. In the rat, the external jugular is a major superficial

Control of pain and distress

It is recognized that any surgical intervention including single housing will have prolonged effects and could affect experimental outcome [21]. This is controlled to a limited extent when experiments are carried out after the rats are brought to a defined state of physiological normalcy. Accordingly, postoperative care is an important part of overall experimental design.

When the catheterization is carried out as described, under aseptic conditions with minimal tissue damage, postoperative

Discussion

A surgical technique for catheterization of the external jugular vein for repeated blood sampling in conscious rats is described in detail with appropriate illustrations. This technique has been successfully used by the investigators for more than a decade. In fact, it would not have been possible to evaluate temporal pattern of hypothalamic–pituitary–adrenal (HPA) axis responses to stressors in conscious rats without the use of this repeated blood sampling technique [2], [8], [14], [16], [27],

Quick procedure

  • 1.

    Prepare sterile catheter and space for aseptic surgery.

  • 2.

    Prepare rat for the surgery.

  • 3.

    Isolate right jugular vein by blunt dissection.

  • 4.

    Introduce the catheter and secure with ligatures.

  • 5.

    Exteriorize through an incision at the nape.

  • 6.

    Fill with gentamicin and occlude the back flow.

  • 7.

    Allow to recover in a temperature controlled chamber.

  • 8.

    House in the home cage after ambulatory.

Essential literature references

[6], [15], [23], [25], [31], [35], [42]

Acknowledgements

The authors thank Dr. Michael J. Huerkamp, DVM, Director, Division of Animal Resources for critically reviewing this manuscript and making helpful suggestions. Supported by MH-50113 and the Emory University Conte Center for the Neuroscience of Mental Retardation MH-58922.

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