Toponomics analysis of drug-induced changes in arachidonic acid-dependent signaling pathways during spinal nociceptive processing.
Multi-Epitope-Ligand-Carthography (MELC) allows consecutive immunohistochemical visualization of up to 100 proteins on the same tissue sample. Subsequent biomathematical analysis of these images allows a quantitative description of changes in protein networks. We used the MELC technology to study the effect of the nonopioid analgesic drug dipyrone on protein network profiles associated with arachidonic acid-dependent signaling pathways. MELC analysis with 31 different fluorescence-labeled tags was used to compare the effect of dipyrone on protein networks in spinal cords of mice with zymosan-induced hyperalgesia, a common model for inflammatory pain. We found that the number of motifs which describe the colocalization of 5-lipoxygenase (5-LO) or 12-LO with other proteins increased disproportionally after dipyrone treatment. Activation of 5-LO and 12-LO induces their translocation to membrane compartments which was also reflected by MELC results. Although no changes in 5-LO or 12-LO expression were seen by Western blot analysis or by immunohistochemistry in spinal cords of dipyrone-treated mice, the activation of both enzymes was verified by determining LO-products. Spinal amounts of 5(S)-hydroxyeicosatetraenoic acid (HETE) and 12(S)-HETE, which are generated by 5-LO and 12-LO, respectively, were significantly increased in spinal cords of dipyrone-treated animals. In primary spinal cord neurons, dipyrone selectively and dose-dependently increased 5(S)-(HETE) and 12(S)-HETE synthesis. Thus, we show for the first time that monitoring protein network profiles by topological proteomic analysis is a useful tool to identify mechanisms of drug actions.
Linke
Bona
B
Pharmazentrum Frankfurt, ZAFES, Institute of Clinical Pharmacology, Klinikum der Goethe-Universität Frankfurt, Germany.
Pierre
Sandra
S
Coste
Ovidiu
O
Angioni
Carlo
C
Becker
Wiebke
W
Maier
Thorsten Jürgen
TJ
Steinhilber
Dieter
D
Wittpoth
Claus
C
Geisslinger
Gerd
G
Scholich
Klaus
K
D000818:Animals; D001092:Arachidonate 12-Lipoxygenase / Q000378:metabolism; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism; D016718:Arachidonic Acid / Q000378:metabolism*; D004177:Dipyrone / Q000494:pharmacology*; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice; D009619:Nociceptors / Q000378:metabolism*; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D020543:Proteome / Q000187:drug effects; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction / Q000187:drug effects*; D013116:Spinal Cord / Q000166:cytology / Q000648:ultrastructure; D018709:Statistics, Nonparametric
D020543:Proteome; D016718:Arachidonic Acid; D004177:Dipyrone; D001092:Arachidonate 12-Lipoxygenase; D001094:Arachidonate 5-Lipoxygenase; D011451:Prostaglandin-Endoperoxide Synthases