title: Mechanisms of apoptosis in embryonic cortical neurons (E6 and E7) in culture involve lipid signalling, protein phosphorylation and caspase activation.
abstract: Cultured embryonic (E7) chick neurons, derived from cerebral hemispheres, underwent apoptosis in response to inhibitors of protein kinase C (staurosporine) and phosphatidylinositol-3-kinase (wortmannin and LY294002), in a dose- and time-dependent manner. This was monitored by loss of cell viability, increased DNA fragmentation, and activation of caspase-3-like activity, all of which were partially reversed by elevating the level of cAMP in the cells with Bt(2)cAMP or (Sp)cAMPS. Further studies revealed that an early step in apoptosis was the formation of ceramide from sphingomyelin, resulting from the activation of a neutral pH sphingomyelinase activity. Thus inhibitors of protein kinase C and phosphatidylinositol-3-kinase increased ceramide levels in the same time-frame as caspase-3 activation and DNA fragmentation. Neurons could also be killed by the addition of either water-soluble C2-ceramide (30 microM) or natural C22/24 ceramide (0.5 microM). In contrast to the apparent protective effect of ser/thr protein phosphorylation, a pro-apoptotic role for tyrosine phosphate phosphorylation was suggested by the ability of protein tyrosine phosphate phosphatase inhibitor, Bis(maltolato)oxovanadium (IV) (BMOV), to induce apoptosis in E7CH neurons. Thus BMOV (25 microM) killed 50% of E7CH neurons and B lymphocytes but not glial cells, or T-lymphocytes, suggesting the existence of a common apoptotic pathway in neurons and B-cells. We conclude that the major pathway for programmed cell death in embryonic chick neurons has many elements in common with that described for other cells but that there may be some unique aspects which can be used to protect embryonic neurons from opioid and other drug-enhanced apoptosis.
authors.affiliation: Department of Pediatrics, University of Chicago, Chicago, IL, USA. dawg@midway.uchicago.edu
authors.forename: G
authors.initials: G
authors.lastname: Dawson
mesh_terms: D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D003994:Bucladesine / Q000494:pharmacology; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism*; D002478:Cells, Cultured; D002518:Ceramides / Q000378:metabolism*; D002540:Cerebral Cortex / Q000166:cytology / Q000196:embryology; D002642:Chick Embryo; D002867:Chromones / Q000494:pharmacology; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005333:Fetus / Q000166:cytology; D009025:Morpholines / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009474:Neurons / Q000166:cytology* / Q000201:enzymology; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010766:Phosphorylation; D011753:Pyrones / Q000494:pharmacology; D015398:Signal Transduction / Q000502:physiology*; D013110:Sphingosine / Q000031:analogs & derivatives / Q000494:pharmacology; D019311:Staurosporine / Q000494:pharmacology; D013873:Thionucleotides / Q000494:pharmacology; D014638:Vanadates / Q000494:pharmacology; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D002518:Ceramides; D002867:Chromones; D004791:Enzyme Inhibitors; D009025:Morpholines; C064769:N-acetylsphingosine; C113227:N-docosanoyl-16-methylheptadecasphing-4-enine; D009419:Nerve Tissue Proteins; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011753:Pyrones; D013873:Thionucleotides; C016957:adenosine-3',5'-cyclic phosphorothioate; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; C074212:bis(maltolato)oxovanadium(IV); D014638:Vanadates; D003994:Bucladesine; D000242:Cyclic AMP; D053148:Caspase 3; D020169:Caspases; D019311:Staurosporine; D013110:Sphingosine; D000077191:Wortmannin