deoxycholic-acid and Neuroblastoma

Acetylcholinesterase (AChE) exists in various molecular forms, depending on alternative splicing of its transcripts and association with structural proteins. Tetramers of the 'tailed' variant (AChE(T)), which are anchored in the cell membrane of neurons by the PRiMA (Proline Rich Membrane Anchor) protein, constitute the main form of AChE in the mammalian brain. In the mouse brain, stress and anticholinesterase inhibitors have been reported to induce expression of the unspliced 'readthrough' variant (AChE(R)) mRNA which produces a monomeric form. To generalize this observation, we attempted to quantify AChE(R) and AChE(T) after organophosphate intoxication in the mouse brain and compared the observed effects with those of stress induced by swimming or immobilization; we also analyzed the effects of heat shock and AChE inhibition on neuroblastoma cells. Active AChE molecular forms were characterized by sedimentation and non-denaturing electrophoresis, and AChE transcripts were quantified by real-time PCR. We observed a moderate increase of the AChE(R) transcript in some cases, both in the mouse brain and in neuroblastoma cultures, but we did not detect any increase of the corresponding active enzyme.

Topics: Acetylcholinesterase; Alternative Splicing; Animals; Blotting, Western; Brain; Cell Line, Tumor; Cholinesterase Inhibitors; Deoxycholic Acid; Detergents; Drug Interactions; Hot Temperature; Male; Mice; Mice, Inbred BALB C; Neuroblastoma; Octoxynol; Proto-Oncogene Proteins c-fos; Receptors, Cholinergic; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Soman; Stress, Physiological; Time Factors

Topics: Adenosine Triphosphatases; Animals; Bucladesine; Cell Differentiation; Cells, Cultured; Clone Cells; Deoxycholic Acid; Hydrogen-Ion Concentration; Insulin; Magnesium; Mice; Neuroblastoma; Potassium; Sodium; Sucrose