Localization of cytosolic phospholipase A2 messenger RNA mainly in neurons in the rat brain.
Ca2(+)-sensitive 85,000 mol. wt cytosolic phospholipase A2 plays an essential role in the selective and stimulus-dependent release of arachidonic acid from membrane phospholipids. Cytosolic phospholipase A2-catalysed lipid mediators including arachidonic acid and its metabolites have been suggested to be involved in a variety of neuronal functions in the CNS. Since the cellular localization of cytosolic phospholipase A2 is still controversial and obscure, we tried an improved method of rapid processing of each specimens and succeeded in obtaining intense signals of cytosolic phospholipase A2 messenger RNA in the normal rat brain by northern blot analysis and in situ hybridization. Northern blot analysis showed the abundant distribution of cytosolic phospholipase A2 messenger RNA in most regions of the brain, with intense signals observed in the pineal gland and pons. Macroautoradiographs prepared after in situ hybridization with three different antisense riboprobes gave essentially similar patterns of localization; significant signals were widely detected in the gray matter of various regions, i.e. the olfactory bulb, cerebral cortex, hippocampus, amygdala, several thalamic and hypothalamic nuclei and cerebellum. Microautoradiographs showed that most of the intense signals were predominant in neurons, and that faint signals were from glial cells and other non-neuronal cells in the choroid plexus, inner surface cells of veins and the leptomeninges. In addition, the cycloheximide treatment increased the cytosolic phospholipase A2 messenger RNA level in the same cell populations originally possessing messenger RNA signals. Predominant expression of cytosolic phospholipase A2 messenger RNA in neurons may provide the basis for the contribution of cytosolic phospholipase A2-catalysed lipid mediators to a variety of neurotransmission and synaptic functions in the CNS.
Kishimoto
K
K
Department of Neuroscience, Osaka Bioscience Institute, Suita-shi, Japan.
Matsumura
K
K
Kataoka
Y
Y
Morii
H
H
Watanabe
Y
Y
D000818:Animals; D001345:Autoradiography; D015152:Blotting, Northern; D001921:Brain / Q000378:metabolism*; D003513:Cycloheximide / Q000494:pharmacology; D003600:Cytosol / Q000201:enzymology*; D017403:In Situ Hybridization; D008297:Male; D008970:Molecular Weight; D009474:Neurons / Q000378:metabolism*; D010741:Phospholipases A / Q000737:chemistry / Q000235:genetics; D054467:Phospholipases A2; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D013116:Spinal Cord / Q000378:metabolism; D014018:Tissue Distribution / Q000502:physiology
D012333:RNA, Messenger; D003513:Cycloheximide; D010741:Phospholipases A; D054467:Phospholipases A2