Enzyme immunoassay for tachykinin-like immunoreactivity in the guinea pig spinal cord.
A solid-phase enzyme immunoassay for quantitation of tachykinin-like immunoreactivity (TK-LI) is presented. Because the antiserum K-12 recognizes various tachykinins, such as neurokinin A (100%), kassinin (103%), eledoisin (51%), neurokinin B (18%), physalaemin (0.7%), and substance P (0.7%), the immunoreactivity detected in this enzyme immunoassay has been termed TK-LI. The assay was performed on 96-well microtiter plates coated with a mouse monoclonal second antibody. After preincubation of soluble neurokinin A or samples and K-12 antiserum for 3 h at room temperature, acetylcholinesterase-labelled neurokinin A was allowed to react overnight at 4 degrees C. Samples were finally incubated with Ellman's reagent for 2 h and the absorbance was measured at 414 nm. The threshold for detection of TK-LI was 2 fmol/well. TK-LI release from guinea pig dorsal spinal cord slices was evoked by capsaicin or high K+ medium. The capsaicin-evoked TK-LI release was increased in the presence of thiorphan, but not in that of captopril.
Renzi
D
D
Department of Clinical Pathophysiology, University of Florence, Italy.
Maillet
S
S
Grassi
J
J
Mantellini
P
P
Santicioli
P
P
Theodorsson
E
E
Surrenti
C
C
Geppetti
P
P
D000110:Acetylcholinesterase; D000818:Animals; D002211:Capsaicin / Q000494:pharmacology; D004600:Eledoisin / Q000032:analysis; D006168:Guinea Pigs; D007124:Immunoenzyme Techniques; D015286:Kassinin / Q000032:analysis; D008297:Male; D015288:Neurokinin A / Q000032:analysis; D015287:Neurokinin B / Q000032:analysis; D010803:Physalaemin / Q000032:analysis; D011188:Potassium / Q000494:pharmacology; D011863:Radioimmunoassay; D013116:Spinal Cord / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D013373:Substance P / Q000032:analysis; D015320:Tachykinins / Q000032:analysis* / Q000378:metabolism; D015244:Thiorphan / Q000494:pharmacology
D015320:Tachykinins; D010803:Physalaemin; D013373:Substance P; D015286:Kassinin; D004600:Eledoisin; D015288:Neurokinin A; D015287:Neurokinin B; D015244:Thiorphan; D000110:Acetylcholinesterase; D011188:Potassium; D002211:Capsaicin