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Microdialysis in Nonhuman Primates
Publication Name:
Current Protocols in Neuroscience
Unit Number:
Unit 7.3
DOI:
10.1002/0471142301.ns0703s00
Online Posting Date:
May, 2001 Abstract
This unit presents a technique that allows for routine repeated microdialysis experiments in the monkey using a sedated preparation or, with further minor modification, in the awake behaving animal. Protocols are provided for construction and in vitro calibration of dialysis probes, obtaining magnetic resonance imaging (MRI) scans of the monkey brain, surgically attaching a guide holder to the skull, and performing postoperative MRI scanning, in vivo dialysis sample collection, anatomical verification of the probes, and neurochemical analysis. An alternate protocol describes microdialysis in awake, behaving monkeys; this requires substantial preparatory work in training the monkeys to sit quietly in a restraining chair and/or to perform a series of behavioral tasks.
Table of Contents
- Unit Introduction
- Basic Protocol 1: Preparation of Dialysis Probe
- Basic Protocol 2: Attachment of Guide Assembly to the Monkey Skull
- Support Protocol 1: Determination of Skull Coordinates by MRI
- Support Protocol 2: Postoperative Verification of Guide Assembly Position by MRI
- Basic Protocol 3: In Vivo Microdialysis
- Support Protocol 3: Anatomical Verification of the Probes
- Alternate Protocol: Modifications for Using In Vivo Microdialysis in the Awake Monkey
- Reagents and Solutions
- Commentary
- Bibliography
- Figures
Materials
Basic Protocol 1: Preparation of Dialysis Probe
Materials
- Epoxy
- Superglue
- 5% to 10% methanol
- Small bags of CaCl
2 - Monkey artificial cerebrospinal fluid (aCSF; appendix
- 3% to 4% formalin (optional)
- MRI scan of monkey brain (see
- Fused silica tubing (o.d. 425 µm, i.d. 320 µm for outer barrel of probe; o.d. 166, i.d. 101 µm for inner barrel of probe)
- Polyethylene (PE) 10 and 50 tubing
- 25-G needle, blunt
- 50-ml syringe
- 35° and 37°C ovens
- Dialysis membrane tubing (e.g., Filtral 20, AN 69, Hospal Medica)
- Dissecting microscope
- 1-ml Hamilton syringe; gas-tight
- Microinfusion pump
- Blue connectors (Bioanalytical Systems)
- 4-O suture thread (optional)
- 700-µl (7 × 40–mm) conical-bottom amber glass vials
- Additional reagents and equipment for analyzing dialysate by HPLC (unit
Basic Protocol 2: Attachment of Guide Assembly to the Monkey Skull
Materials
- 6- to 12.5-kg rhesus monkey (Macaca mulatta)
- Ketamine
- Isoflurane
- Ringer's solution
- Bone cement
- Guide holder
- Intubation tube
- Venous catheter
- i.v. setup
- Vital sign monitor and pads
- MRI-compatible stereotaxic instrument, equipped with manipulators
- 37°C heating blanket
- Surgical instruments for aseptic neurosurgery
- Guide block, plastic cap, and screws
- Marking pen, sterile
- Drill, hand-held or dental, 14 to 20 mm × 7 to 9 mm
- Rongeurs (optional)
- Plastic I-shaped anchors
- Additional reagents and equipment for determining skull coordinates by MRI (see
Support Protocol 1: Determination of Skull Coordinates by MRI
Additional Materials (also see Basic Protocol 2 )
- Xylazine
- Container for transporting monkey
- GE/Signa 1.5 T MRI scanner or equivalent
Support Protocol 2: Postoperative Verification of Guide Assembly Position by MRI
Additional Materials (also see Basic Protocol 2 and Support Protocol 1 )
- Vitamin E capsules
- Soft clay
- Fused silica tubing (o.d. 425 µm, i.d. 320 µm)
Basic Protocol 3: In Vivo Microdialysis
Materials
- Monkey artificial cerebrospinal fluid (aCSF; see appendix
- Ketamine hydrochloride
- Monkey with attached dialysis probe guide assembly (see
- Isoflurane
- Ophthalmic antibiotic ointment
- Dialysis probes (see
- 700-µl (7 × 40–mm) conical-bottom amber glass vials
- 1-ml Hamilton syringe, gas-tight
- Microinfusion pump
- MRI-compatible stereotaxic instrument
- 25-G needle
- 25-G stainless steel tubing (o.d. 0.020 in., i.d. 0.010 in.; Hypo tube, Small Parts)
- Blue connectors (Bioanalytical Systems)
- Polyethylene (PE) 10 tubing
- 4-O suture thread (optional)
- Additional reagents and equipment for performing aseptic neurosurgery (see
Alternate Protocol: Modifications for Using In Vivo Microdialysis in the Awake Monkey
Additional Materials (also see Basic Protocol 3 )
- Primate chair
- Stainless steel post
- Steel straps
- Bone cement
- No. 8 screws
- Steel rod
- Crossing rod
- 4 × 8–in. thermoplastic sheet
- Nuts
Looking for materials? Suppliers Guide
Figures
-
Figure 7.3.1Diagramatic representations of the guide assembly and dialysis probe. (A) Spacing of the guide holes into six rows of 15 holes (<1 mm in diameter) with the holes spaced 1 mm apart. (B) View of the guide holder, guide block, and cap which attach together with plastic screws. (C) Schematic view of the dialysis probe: 1, AN 69 dialysis membrane; 2, fused silica inner barrel; 3, fused silica outer barrel; 4, plastic sleeve tubing (PE 50); 5, fused silica barrel of outlet tube; 6, inlet and outlet tubing. (D) Example of probe positioned through the guide assembly. -
Figure 7.3.2MRI scans of two coronal sections of monkey brain at the level of the caudate nucleus. The white arrows indicate positions of vitamin E–filled silica tubes in the guide assembly during the MRI scan. The position of the guides confirms placement of the guide holder above the appropriate area, with several guide holes able to direct dialysis probes into the caudate nucleus. -
Figure 7.3.3Photograph of the sedate monkey during a dialysis session. The monkey is placed in a stereotaxic head holder with the pump, collection vials, and liquid switches in close proximity to the head and probes. In this experiment, there are six probes positioned in the brain: two in the parietal cortex, two in the prefrontal cortex, and two in the caudate nucleus.
Literature Cited
| Literature Cited | |
| Kolachana, B.S., Saunders, R.C., and Weinberger, D.R. 1994. An improved methodology for routine in vivo microdialysis in nonhuman primates. J. Neurosci. Methods 55:1-6. | |
| Kolachana, B.S., Saunders, R.C., and Weinberger, D.R. 1996. In vivo characterization of extracellular GABA release in the caudate nucleus and prefrontal cortex of the rhesus monkey. Synapse 25:1-8. | |
| NIH (National Institutes of Health). 1985. Guide for the Care and Use of Laboratory Animals. NIH Publication no. 85-23. NIH, Bethesda, Md. | |
| Saunders, R.C., Aigner, T.G., and Frank, J.A. 1990 Magnetic resonance imaging of the rhesus monkey brain: Use for stereotactic neurosurgery. Exp. Brain Res. 81:443-446. | |
| Saunders, R.C., Kolachana, B.S., and Weinberger, D.R. 1993. Local pharmacological manipulation of dopamine in prefrontal cortex and caudate in the rhesus monkey: An in vivo microdialysis study. Exp. Brain Res. 98:44-52. | |
| Wang, J., Skirboll, S., Aigner, T.G., Saunders, R.C., Hsiao, J., and Bankiewicz, K.S. 1990. Methodology of microdialysis of neostriatum in hemiparkinsonian nonhuman primates. Exp. Neurol. 110:181-186. | |
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