Translocation of cytochrome c during cerebellar degeneration in Lurcher and weaver mutant mice.

Cytochrome c translocation from the inner mitochondrial membrane into the cytosol is the initial step of the intrinsic apoptotic pathway. As no evidence was ever presented for cytochrome c translocation during cerebellar degeneration in Lurcher (Lc/+) and weaver (wv/wv) mutant mice, we searched for the presence of such a process in cerebellar homogenates of mutant and wild-type mice from postnatal day (P)1 to P56. Here we present the first documented time course of cytochrome c translocation spanning the entire period of neurodegeneration in both mutant types. We identified cytochrome c with Western blotting and monitored cell loss in the cerebellum with Calbindin D-28k immunohistochemistry, Nissl-staining and morphometry. No cytochrome c translocation was ever detected in wild-types at any age investigated. Translocated cytochrome c appeared between P13 and P21 in Lc/+ and between P5 and P6 in wv/wv. These two intervals precisely coincide with the respective periods of maximal neuronal death in the cerebellum. Secondary translocation was also observed at a later stage between P42 and P49 in Lc/+ and from P22 onwards in wv/wv. Since no substantial neuronal loss has ever been observed in Lc/+ and wv/wv mutants at these postnatal ages, the delayed translocation may correspond to cytochrome c of extraneuronal, presumably glial origin. Observations of an increased expression of glial fibrillary acidic protein and sustained remodeling of the astrocytic network in the cerebellum of both mutants, long after the cessation of neuronal death make this assumption rather plausible.