TY - JOUR
T1 - Multiple apoptogenic proteins are involved in the nuclear translocation of Apoptosis Inducing Factor during transient focal cerebral ischemia in rat
AU - Chaitanya, Ganta Vijay
AU - Babu, Phanithi Prakash
N1 - Funding Information:
Funding from DST, DBT and ICMR, India is gratefully acknowledged. The authors thank Dr. Abira Sarkar UoH, India, for HSP-70 antibody and Dr. Siva Kumar for Cy5 conjugated rabbit secondary antibody. The authors acknowledge Mr. Murty, Central Instruments Laboratory for his help in confocal microscopy. Financial support to Ganta Vijay Chaitanya in the form of SRF from UCG is gratefully acknowledged.
PY - 2008/12/30
Y1 - 2008/12/30
N2 - Apoptosis Inducing Factor is a mitochondrial protein which upon translocation to nucleus causes large scale DNA fragmentation. The stimulus for the cytosolic release and nuclear translocation for this protein still remains to be understood. The role of calpains, cathepsin-b, Poly ADP (ribose) Polymerase and granzyme-b in the nuclear translocation of AIF has been investigated in the pathology of cerebral ischemia. Calpains, cathepsin-b and PARP-1 which were mostly confined to cytosol, lysosomes and nucleus respectively were found to be elevated in the mitochondrial fraction interacting with AIF in the western blot analysis and double immunofluorescence analysis. Western blot and immunohistochemical analysis revealed elevated levels of granzyme-b secreted by cytotoxic T lymphocytes and natural killer cells in the infarct of ischemic mouse brain. Co-immunoprecipitation revealed and western blot analysis the interaction and break down of Heat Shock Protein-70 an endogenous inhibitor of AIF into signature fragments by granzyme-b facilitating the nuclear translocation of AIF. Break down of HSP-70 correlated with the nuclear translocation of AIF observed in western and immunohistochemical analysis. These results indicate that multiple proteases were involved in the nuclear translocation of AIF during the pathology of cerebral ischemia.
AB - Apoptosis Inducing Factor is a mitochondrial protein which upon translocation to nucleus causes large scale DNA fragmentation. The stimulus for the cytosolic release and nuclear translocation for this protein still remains to be understood. The role of calpains, cathepsin-b, Poly ADP (ribose) Polymerase and granzyme-b in the nuclear translocation of AIF has been investigated in the pathology of cerebral ischemia. Calpains, cathepsin-b and PARP-1 which were mostly confined to cytosol, lysosomes and nucleus respectively were found to be elevated in the mitochondrial fraction interacting with AIF in the western blot analysis and double immunofluorescence analysis. Western blot and immunohistochemical analysis revealed elevated levels of granzyme-b secreted by cytotoxic T lymphocytes and natural killer cells in the infarct of ischemic mouse brain. Co-immunoprecipitation revealed and western blot analysis the interaction and break down of Heat Shock Protein-70 an endogenous inhibitor of AIF into signature fragments by granzyme-b facilitating the nuclear translocation of AIF. Break down of HSP-70 correlated with the nuclear translocation of AIF observed in western and immunohistochemical analysis. These results indicate that multiple proteases were involved in the nuclear translocation of AIF during the pathology of cerebral ischemia.
KW - Apoptosis Inducing Factor
KW - Calpain
KW - Cathepsin-b
KW - Cerebral ischemia
KW - Granzyme-b
KW - Heat Shock Protein-70
KW - Poly ADP (ribose) Polymerase
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U2 - 10.1016/j.brainres.2008.09.075
DO - 10.1016/j.brainres.2008.09.075
M3 - Article
C2 - 18938146
AN - SCOPUS:57249103355
SN - 0006-8993
VL - 1246
SP - 178
EP - 190
JO - Brain Research
JF - Brain Research
ER -