Recent advances in liquid chromatography combined with tandem MS systems made possible the detection of novel bioactive acid derivatives as well as the simultaneous monitoring of the known members of a given sub-family of endocannabinoid-like molecules. We have previously developed sensitive and specific "targeted lipidomics" methods using high resolution LC-ESI-IT-ToF (Liquid Chromatography-ElectroSpray Ionization-Ion Trap-Time of Flight) mass spectrometry for the simultaneous identification and quantification in tissues of trace amounts either of cyclooxygenase-2 (COX-2) derivatives of anandamide and 2-AG or of endocannabinoids/endovanilloids such as the N-acyl-dopamines. The application of these techniques should now make possible the study of the presence and regulation of these two poorly understood classes of lipid mediators in CNS diseases. For example, we applied one of these methods for the investigation on the occurrence and role of prostaglandin glycerol esters in an experimental model of Huntington's disease (Sagredo et al., 20th Annual meeting of the ICRS, Lund, Sweden, 2010). Here we report the results of two different studies on prostaglandin-ethanolamides (prostamides) and N-arachidonoyl-dopamine (NADA). In the first study, we investigated the possible formation of prostamides during knee inflammation induced by kaolin+carrageenan in mice. This condition was accompanied by elevation of the spinal levels of prostamide F2?, which was shown to participate in inflammation-evoked spinal hyperexcitability. Importantly, the elevation of prostamide F2? levels was counteracted by concomitant inhibition of COX-2. In the second study, we demonstrated for the first time the activity-dependent biosynthesis of the endocannabinoid/endovanilloid NADA in slices of the rat substantia nigra compacta. Both high K+ and group I glutamate receptor (mGluR) stimulation led to elevation of NADA levels in a manner sensitive to carbidopa. These findings indicate that the developed targeted lipidomics procedures are suitable for studies on the levels of N-acyl-dopamines and endocannabinoid COX-2 derivatives in nervous tissues, and represent the first reports of the detection of prostamide F2? in the CNS, and of the activity-dependent production of NADA.
Endocannabinoidomics: targeted lipidomics methods for the study of endocannabinoid-related molecules in the CNS
T Bisogno;V Di Marzo
2011
Abstract
Recent advances in liquid chromatography combined with tandem MS systems made possible the detection of novel bioactive acid derivatives as well as the simultaneous monitoring of the known members of a given sub-family of endocannabinoid-like molecules. We have previously developed sensitive and specific "targeted lipidomics" methods using high resolution LC-ESI-IT-ToF (Liquid Chromatography-ElectroSpray Ionization-Ion Trap-Time of Flight) mass spectrometry for the simultaneous identification and quantification in tissues of trace amounts either of cyclooxygenase-2 (COX-2) derivatives of anandamide and 2-AG or of endocannabinoids/endovanilloids such as the N-acyl-dopamines. The application of these techniques should now make possible the study of the presence and regulation of these two poorly understood classes of lipid mediators in CNS diseases. For example, we applied one of these methods for the investigation on the occurrence and role of prostaglandin glycerol esters in an experimental model of Huntington's disease (Sagredo et al., 20th Annual meeting of the ICRS, Lund, Sweden, 2010). Here we report the results of two different studies on prostaglandin-ethanolamides (prostamides) and N-arachidonoyl-dopamine (NADA). In the first study, we investigated the possible formation of prostamides during knee inflammation induced by kaolin+carrageenan in mice. This condition was accompanied by elevation of the spinal levels of prostamide F2?, which was shown to participate in inflammation-evoked spinal hyperexcitability. Importantly, the elevation of prostamide F2? levels was counteracted by concomitant inhibition of COX-2. In the second study, we demonstrated for the first time the activity-dependent biosynthesis of the endocannabinoid/endovanilloid NADA in slices of the rat substantia nigra compacta. Both high K+ and group I glutamate receptor (mGluR) stimulation led to elevation of NADA levels in a manner sensitive to carbidopa. These findings indicate that the developed targeted lipidomics procedures are suitable for studies on the levels of N-acyl-dopamines and endocannabinoid COX-2 derivatives in nervous tissues, and represent the first reports of the detection of prostamide F2? in the CNS, and of the activity-dependent production of NADA.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
