SpringerProtocols: Abstract: 13. In Situ Brain Perfusion Technique

13. In Situ Brain Perfusion Technique

By: Quentin R. Smith³, David D. Allen³

Abstract

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Many techniques have been developed to study transport across the blood-brain barrier (BBB). One that offers particular advantage is the in situ brain perfusion technique. The primary objective of the in situ brain perfusion technique is to take over the circulation to the brain via direct infusion of artificial blood, plasma, or saline into the heart or major vessels leading to the brain, such as the carotid arteries (1–4). The solute of interest is mixed into the perfusion medium at a known concentration and delivered to the brain via the perfusion fluid for a defined interval. At preselected times, the perfusion is stopped and the amount of solute in brain is determined. From these measurements, the kinetics of brain uptake are analyzed and appropriate transport or permeability constants calculated (5).

Affiliation(s): (3) Department of Pharmaceutical Sciences, Texas Tech University Health Sciences Center, Amarillo, TX

Book Title: The Blood-Brain Barrier: Biology and Research Protocols

Series: Methods in Molecular Medicine | Volume: 89 | Pub. Date: Aug-06-2003 | Page Range: 209-218 | DOI: 10.1385/1-59259-419-0:209

Subject: Neuroscience

References

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1.	Takasato, Y., Rapoport, S. I., and Smith, Q. R. (1982) A new method to determine cerebrovascular permeability in the anesthetized rat. Soc. Neurosci. Abstr. 8, 850.

2.	Takasato Y., Rapoport S. I., and Smith Q. R. (1984) An in situ brain perfusion technique to study cerebrovascular transport in the rat. Am. J. Physiol. 247, H484–H493.

3.	Smith, Q. R. (1996) Brain perfusion systems for studies of drug uptake and metabolism in the central nervous system. Pharm. Biotechnol. 8, 285–307.

4.	Smith, Q. R., and Nagura, H. (2001) Fatty acid uptake and incorporation in brain. J. Mol. Neurosci. 16, 167–172.

5.	Smith, Q.R. (1989) Quantitation of blood-brain barrier permeability. In: Implications of the Blood-Brain Barrier and Its Manipulation. Vol. 1. Neuwelt, E. A., ed. Plenum Press, New York, pp 85–118.

6.	Hokari, M., Wu, H. Q., Schwarcz, R., and Smith, Q. R. (1996) Facilitated brain uptake of 4-chlorokynureine and conversion to 7-chlorokynurenic acid. NeuroReport 8, 15–18.

7.	Simpson, I. A., Appel, N. M., Hokari, M., et al. (1999) Blood-brain barrier glucose transporter: Effects of hypoglycemia and hyperglycemia revisited. J. Neurochem. 72, 238–247.

8.	Momma, S., Aoyagi, M., Rapoport, S. I., and Smith, Q. R. (1987) Phenylalanine transport across the blood-brain barrier as studied with the in situ brain perfusion technique. J. Neurochem. 48, 1291–1300.

9.	Takada, Y., Vistica, D. T., Greig, N. H., Purdon, D., Rapoport, S. I., and Smith, Q. R. (1992) Rapid high-affinity transport of a chemotherapeutic amino acid across the blood-brain barrier. Cancer Res. 52, 2191–2196.

10.	Allen, D. D., and Smith, Q. R. (2001) Characterization of the blood-brain barrier choline transporter using the in situ rat brain perfusion technique. J. Neurochem. 76, 1032–1041.

11.	Triguero, D., Buciak, J., and Pardridge, W. M. (1990) Capillary depletion method for quantification of blood-brain barrier transport of circulating peptides and plasma proteins, J. Neurochem. 54, 1882–1888.

12.	Rabin, O., Hegedus, L., Bourre, J. M., and Smith, Q. R. (1993) Rapid brain uptake of manganese(II) across the blood-brain barrier. J. Neurochem. 61, 509–517.

13.	Thompson, A. M., Robertson, R. C, and Bauer, T. A. (1968) A rat head-perfusion technique developed for the study of brain uptake of materials, J. Appl. Physiol. 24, 407–411.

14.	Greenwood, J., Luthert, P. J., Pratt, O. E., and Lantos, P. L. (1985) Maintenance of the integrity of the blood-brain barrier in the rat during an in situ saline-based perfusion, Neurosci. Lett. 56, 223–227.

15.	Luthert, P. J., Greenwood, J., Pratt, O. E., and Lantos, P. L. (1987) The effect of metabolic inhibitor upon the properties of the cerebral vasculature during whole head saline perfusion of the rat, Quart. J. Exp. Physiol. 72, 129–141.

16.	Allen, D. D., and Smith, Q. R. (1999) Blood-brain barrier choline transport in the senescent rat. Neuroscience Letters. 277, 198–202.

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