Bax Deletion Further Orders the Cell Death Pathway in Cerebellar Granule Cells and Suggests a Caspase-independent Pathway to Cell Death

• DOI: 10.1083/jcb.139.1.205
• Publication Date: 2002-07-26T16:47:50Z
• Authors: Timothy M. Miller; Krista L. Moulder; C. Michael Knudson; Douglas J. Creedon; Mohanish Deshmukh; Stanley J. Korsmeyer; Eugene M. Johnson
• Keywords: Granule (geology)

Dissociated cerebellar granule cells maintained in medium containing 25 mM potassium undergo an apoptotic death when switched to with 5 potassium. Granule from mice which Bax, a proapoptotic Bcl-2 family member, had been deleted, did not apoptosis potassium, yet excitotoxic cell response stimulation 30 or 100 μM NMDA. Within 2 h after switching K+, both wild-type and Bax-deficient decreased glucose uptake <20% of control. Protein synthesis also rapidly 50% control within 12 Both Bax −/− neurons increased mRNA levels c-jun, caspase 3 (CPP32) phosphorylation the transactivation domain c-Jun K+ deprivation. Wild-type cleavage DEVD–aminomethylcoumarin (DEVD-AMC), fluorogenic substrate for caspases 2, 3, 7; contrast, cleave DEVD-AMC. These results place BAX downstream metabolic changes, changes levels, c-Jun, upstream activation indicate that is required apoptotic, but excitotoxic, death. In cells, Boc-Asp-FMK ZVAD-FMK, general inhibitors caspases, blocked DEVD-AMC increase TdT-mediated dUTP nick end labeling (TUNEL) positivity. However, these only marginal effect on preventing death, suggesting caspase-independent pathway cells.