RESUMEN
We studied the distribution of 12-lipoxygenase mRNA in glial cells. First, mRNA was detected from cellular extracts by soluble-phase reverse transcriptase-polymerase chain reaction (RT-PCR). Taking into account that cell culture populations could not be 100% homogeneous, we then developed, for the first time, an in situ RT-PCR combined with immunocytochemistry with cell specific markers. Using this procedure we showed that 12-lipoxygenase mRNA was expressed both in mature oligodendrocytes and astrocytes.
Asunto(s)
Araquidonato 12-Lipooxigenasa/biosíntesis , Astrocitos/enzimología , Oligodendroglía/enzimología , ARN Mensajero/biosíntesis , Animales , Animales Recién Nacidos , Secuencia de Bases , Células Cultivadas , Inmunohistoquímica , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , ADN Polimerasa Dirigida por ARN , Ratas , Fijación del TejidoRESUMEN
12-hydroxy 5,8,14-cis 10-trans eicosatetraenoic acid (12-HETE) and its derivatives are the principal lipoxygenase (Lox) products of the mammalian brain. These metabolites have been proposed to play a key role as second messengers in synaptic transmission and might function as retrograde messengers in learning and memory processes: the long-term potentiation. The exact source(s) of 12-HETE and neuronal implication have not been definitively established. The present work was therefore designed to study 12-Lox mRNA expression in neural cell cultures. Detection of this mRNA from cellular extract was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR) and localization in neurons by in situ RT-PCR. These results argue for 12-Lox neuronal production.