Disulfiram administration affects substance P-like immunoreactive and monoaminergic neural systems in rodent brain.
The biosynthetic enzyme peptidylglycine alpha-amidating monooxygenase catalyzes the formation of a variety of biologically active alpha-amidated peptides from respective COOH-terminal glycine-extended peptide precursors. Peptidylglycine alpha-amidating monooxygenase activity is dependent on copper, ascorbate, and molecular oxygen and is inhibited by the relatively selective copper chelator N,N-diethyldithiocarbamate or its disulfide dimer disulfiram (Antabuse). In the present study, chronic disulfiram treatment (100 mg/kg/day, for 12-25 days) resulted in significant changes in several neurochemical parameters in the mouse central nervous system, including levels of substance P-like, unamidated substance P-Gly-like, and protease-generated substance P-Gly-Lys-like immunoreactivities (SP-LI, SP-G-LI, and SP-G-K-LI, respectively). Combined high performance liquid chromatography/radioimmunoassay analyses of the extracted SP-LI, SP-G-LI, and SP-G-K-LI species indicated very similar chromatographic and immunochemical behavior as demonstrated for chemically authentic peptide standards. Additionally, changes in levels of monoamines and their metabolites were observed after drug administration. Complementary immunohistochemical analyses using affinity-purified anti-SP-G sera localized these drug-induced changes in levels of immunoreactive unamidated precursor to neural elements that normally express SP. As a functional corollary to alterations in neurochemical parameters, we observed significant disulfiram-induced increases in pain thresholds, potentiated by capsaicin treatment. Overall, our results indicate that the observed changes in steady state levels of immunoreactive SP and of the immature COOH-terminal extended forms of SP may reflect compensatory biosynthetic and posttranslational processing events in SP-containing neural systems after pharmacological challenge.
Department of Anesthesiology, Tufts University School of Medicine, Boston, Massachusetts 02111.
J E
JE
Marchand
K
K
Hershman
M S
MS
Kumar
M L
ML
Thompson
R M
RM
Kream
D000595:Amino Acid Sequence; D000818:Animals; D015306:Biogenic Monoamines / Q000378:metabolism*; D001921:Brain / Q000187:drug effects / Q000378:metabolism*; D002211:Capsaicin / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D004221:Disulfiram / Q000494:pharmacology*; D004357:Drug Synergism; D007033:Hypothalamus, Middle / Q000378:metabolism; D007150:Immunohistochemistry; D008526:Medulla Oblongata / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D010147:Pain Measurement; D011149:Pons / Q000378:metabolism; D011301:Preoptic Area / Q000378:metabolism; D011863:Radioimmunoassay; D013116:Spinal Cord / Q000378:metabolism; D013373:Substance P / Q000378:metabolism*
D015306:Biogenic Monoamines; D013373:Substance P; D002211:Capsaicin; D004221:Disulfiram