Neuronal cell death can occur through two principal mechanisms: necrosis and apoptosis. Caspase-family proteases are the major effectors of the apoptotic program family. Pathological activation of Caspases has been demonstrated in models of brain injury, including ischemia, neurodegeneration, and neuroinflammation, making these proteases targets for potential drug discovery. Because the human genome contains 11 Caspases, however, a challenge is defining which of these proteases are critical for neuronal cell death in vivo. Caspase-8 is the apical protease in a cell death pathway activated by Tumor Necrosis Factor (TNF) family cytokine receptors. Circumstantial evidence has implicated Caspase-8 in neuronal cell death during ischemia and certain neurodegenerative conditions. Calpains are also intracellular cysteine proteases, which are activated by elevations in intracellular Ca 2+ and which have been implicated in neuronal cell death in models of ischemia and some neurodegenerative diseases. Interestingly, Calpains and Caspases cleave many of the same substrates, implying common or redundant cell death mechanisms. The human genome contains 12 Calpain-family proteases, with Calpains 1 and 2 being most clearly implicated in neuronal cell death. Attempts to directly evaluate the importance of Caspase-8 and Calpains in neuronal cell death in vivo have been hampered by the embryonic lethality that occurs upon knocking-out these genes in mice. We therefore propose to use cre-lox technology for achieving neuron-specific ablation in mice of the genes encoding Caspase-8 and the common subunit shared by Calpains 1 and 2 (capn4). These animals and cells derived from them will then be used for determining the importance of these proteases for neuronal cell death induced by cerebral ischemia and in certain neurodegenerative diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS036821-08
Application #
7002667
Study Section
Molecular, Cellular and Developmental Neurosciences 2 (MDCN)
Program Officer
Hicks, Ramona R
Project Start
1999-03-01
Project End
2006-12-31
Budget Start
2006-01-01
Budget End
2006-12-31
Support Year
8
Fiscal Year
2006
Total Cost
$452,242
Indirect Cost
Name
Sanford-Burnham Medical Research Institute
Department
Type
DUNS #
020520466
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Krajewski, Stan Stanislaw; Tsukamoto, Michelle M; Huang, Xianshu et al. (2015) Nonstripping ""Rainbow"" and Multiple Antigen Detection (MAD) Western Blotting. Methods Mol Biol 1314:287-301
Krajewski, Stan; Wang, Jeffrey; Khan, Tashmia et al. (2015) Image analysis algorithms for immunohistochemical assessment of cell death. Methods Mol Biol 1254:181-96
Krajewska, Maryla; You, Zerong; Rong, Juan et al. (2011) Neuronal deletion of caspase 8 protects against brain injury in mouse models of controlled cortical impact and kainic acid-induced excitotoxicity. PLoS One 6:e24341
Krajewski, Stan; Huang, Xianshu; Krajewska, Maryla (2009) Multiple antigen detection (MAD) western blotting. Methods Mol Biol 536:473-81
Krajewska, Maryla; Smith, Layton H; Rong, Juan et al. (2009) Image analysis algorithms for immunohistochemical assessment of cell death events and fibrosis in tissue sections. J Histochem Cytochem 57:649-63
Krajewski, Stan (2009) ""Rainbow"" western blotting. Methods Mol Biol 536:463-72
Benchoua, Alexandra; Trioulier, Yael; Diguet, Elsa et al. (2008) Dopamine determines the vulnerability of striatal neurons to the N-terminal fragment of mutant huntingtin through the regulation of mitochondrial complex II. Hum Mol Genet 17:1446-56
Krajewska, Maryla; Banares, Steven; Zhang, Eric E et al. (2008) Development of diabesity in mice with neuronal deletion of Shp2 tyrosine phosphatase. Am J Pathol 172:1312-24
Konig, Hans-Georg; Rehm, Markus; Gudorf, Daniel et al. (2007) Full length Bid is sufficient to induce apoptosis of cultured rat hippocampal neurons. BMC Cell Biol 8:7
Kim, Hyung-Ryong; Chae, Han-Jung; Thomas, Michael et al. (2007) Mammalian dap3 is an essential gene required for mitochondrial homeostasis in vivo and contributing to the extrinsic pathway for apoptosis. FASEB J 21:188-96

Showing the most recent 10 out of 38 publications