Measurement of cholecystokinin release in vivo in the rat spinal dorsal horn.
The microdialysis technique, used to monitor extracellular levels of transmitter substances in the central nervous system of laboratory animals as a reflection of transmitter release, is based on the ability of neurotransmitters to diffuse in the extracellular fluid from the site of release and to cross a semipermeable dialysis membrane. Even though the surgical procedure is not very complicated, the detection of released substances in the recovered dialysate may be difficult. Especially, the measurement of neuropeptide release is limited by the low extracellular concentration and of low recovery as compared to, for example, monoamines. Thus, for example, cholecystokinin (CCK), which is the most abundant neuropeptide in the central nervous system, is found at concentrations that are several orders of magnitude lower than those of classical transmitters. Therefore a highly sensitive detection method is of utmost importance. In the dorsal horn of the spinal cord CCK is found mainly in interneurons and in terminals of descending fibers. CCK seems to be involved in nociceptive transmission and CCK attenuates morphine-induced antinociception. We here describe in vivo microdialysis in the lumbar dorsal horn of the rat with subsequent quantification of the level of CCK-like immunoreactivity (-LI) by a highly sensitive radioimmunoassay.
Gustafsson
H
H
Karolinska Institute, Department of Physiology and Pharmacology, Division of Pharmacological Pain Research, S-171 77, Stockholm, Sweden.
de Araujo Lucas
G
G
Schött
E
E
Stiller
C O
CO
Alster
P
P
Wiesenfeld-Hallin
Z
Z
Brodin
E
E
D000344:Afferent Pathways; D000818:Animals; D002766:Cholecystokinin / Q000032:analysis / Q000378:metabolism*; D008297:Male; D017551:Microdialysis / Q000379:methods*; D011188:Potassium / Q000494:pharmacology; D011863:Radioimmunoassay; D051381:Rats; D017207:Rats, Sprague-Dawley; D012844:Sincalide / Q000032:analysis; D013116:Spinal Cord / Q000378:metabolism*
D002766:Cholecystokinin; D012844:Sincalide; D011188:Potassium