Asunto(s)
Enfermedad de Alzheimer/líquido cefalorraquídeo , Proteínas del Líquido Cefalorraquídeo/análisis , Proteínas del Tejido Nervioso/análisis , Proteínas del Tejido Nervioso/líquido cefalorraquídeo , Enfermedad de Parkinson/líquido cefalorraquídeo , Proteómica/métodos , Enfermedad de Alzheimer/diagnóstico , Enfermedad de Alzheimer/fisiopatología , Apolipoproteínas E/análisis , Apolipoproteínas E/líquido cefalorraquídeo , Biomarcadores/análisis , Biomarcadores/líquido cefalorraquídeo , Encéfalo/metabolismo , Encéfalo/fisiopatología , Complemento C4a/análisis , Complemento C4a/líquido cefalorraquídeo , Diagnóstico Diferencial , Proteínas del Ojo/análisis , Proteínas del Ojo/líquido cefalorraquídeo , Humanos , Complejos Multienzimáticos/análisis , Complejos Multienzimáticos/líquido cefalorraquídeo , Factores de Crecimiento Nervioso/análisis , Factores de Crecimiento Nervioso/líquido cefalorraquídeo , Enfermedad de Parkinson/diagnóstico , Enfermedad de Parkinson/fisiopatología , Fosfodiesterasa I/análisis , Fosfodiesterasa I/líquido cefalorraquídeo , Hidrolasas Diéster Fosfóricas , Valor Predictivo de las Pruebas , Pirofosfatasas/análisis , Pirofosfatasas/líquido cefalorraquídeo , Serpinas/análisis , Serpinas/líquido cefalorraquídeo , Superóxido Dismutasa/análisis , Superóxido Dismutasa/líquido cefalorraquídeo , Superóxido Dismutasa-1 , Regulación hacia Arriba/fisiologíaAsunto(s)
Técnicas para Inmunoenzimas/métodos , Complejos Multienzimáticos/líquido cefalorraquídeo , Complejos Multienzimáticos/inmunología , Fosfodiesterasa I/líquido cefalorraquídeo , Fosfodiesterasa I/inmunología , Pirofosfatasas/líquido cefalorraquídeo , Pirofosfatasas/inmunología , Humanos , Hidrolasas Diéster FosfóricasRESUMEN
We previously reported that nerve injury-induced neuropathic pain and its underlying mechanisms are initiated by lysophosphatidic acid. In the present study, by measuring cell-rounding in a biological assay using lysophosphatidic acid 1 receptor-expressing B103 cells, we evaluated the molecular mechanism underlying lysophosphatidic acid biosynthesis following intense stimulation of primary afferents. Lysophosphatidic acid production was induced by treatment of spinal cord slices with capsaicin (10 microM), an intense stimulator of primary afferents, in the presence of recombinant autotaxin, but not in its absence. Lysophosphatidic acid was also induced by combination treatment of slices with high doses (10 and 30 microM) of substance P and NMDA, but not by other combinations of substance P, NMDA, calcitonin gene-related peptide and alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (30 microM each) in the presence of recombinant autotaxin. We also found that following neurokinin 1 and NMDA receptor activation, activation of both cytosolic phospholipase A(2) and calcium-independent intracellular phospholipase A(2) signalling pathways through protein kinase C and mitogen-activated protein/extracellular signal-regulated kinase activation and intracellular calcium elevation were required for lysophosphatidic acid production. These findings suggest that simultaneous intense stimulation of neurokinin 1 and NMDA receptors in the spinal dorsal horn triggers lysophosphatidic acid production from lysophosphatidylcholine through extracellular autotaxin.
Asunto(s)
Lisofosfatidilcolinas/metabolismo , Lisofosfolípidos/metabolismo , Complejos Multienzimáticos/metabolismo , Fosfodiesterasa I/metabolismo , Pirofosfatasas/metabolismo , Receptores de N-Metil-D-Aspartato/fisiología , Receptores de Neuroquinina-1/fisiología , Médula Espinal/metabolismo , Vías Aferentes/fisiología , Análisis de Varianza , Animales , Péptido Relacionado con Gen de Calcitonina/farmacología , Capsaicina/farmacología , Línea Celular Transformada , Relación Dosis-Respuesta a Droga , Agonistas de Aminoácidos Excitadores/farmacología , Técnicas In Vitro , Lisofosfolípidos/farmacología , Ratones , Complejos Multienzimáticos/líquido cefalorraquídeo , N-Metilaspartato/farmacología , Fosfodiesterasa I/líquido cefalorraquídeo , Fosfolipasas A2/metabolismo , Hidrolasas Diéster Fosfóricas , Proteína Quinasa C/metabolismo , Pirofosfatasas/líquido cefalorraquídeo , Receptores del Ácido Lisofosfatídico/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Médula Espinal/efectos de los fármacos , Sustancia P/farmacología , Ácido alfa-Amino-3-hidroxi-5-metil-4-isoxazol Propiónico/farmacologíaRESUMEN
Abstract Cerebrospinal fluid (CSF) induced neurite retraction of differentiated PC12 cells; the action was observed in 15 min (a rapid response) and the activity further increased until 6 h (a long-acting response) during exposure of CSF to the cells. The CSF action was sensitive to monoglyceride lipase and diminished by homologous desensitization with lysophosphatidic acid (LPA) and by pretreatment with an LPA receptor antagonist Ki16425. Although fresh CSF contains LPA to some extent, the LPA content in the medium was increased during culture of PC12 cells with CSF. The rapid response was mimicked by exogenous LPA, and a long-acting response was duplicated by a recombinant autotaxin, lysophospholipase D (lyso-PLD). Although the lyso-PLD substrate lysophosphatidylcholine (LPC) was not detected in CSF, lyso-PLD activity and an approximately 120-kDa autotaxin protein were detected in CSF. On the other hand, LPC but not lyso-PLD activity was detected in the conditioned medium of a PC12 cell culture without CSF. Among neural cells examined, leptomeningeal cells expressed the highest lyso-PLD activity and autotaxin protein. These results suggest that leptomeningeal cells may work as one of the sources for autotaxin, which may play a critical role in LPA production and thereby regulate axonal and neurite morphological change.
