Rotation Projects to Examine the Molecular Mechanism of Apoptosis in Neurons

We are interested in understanding the molecular mechanism by which neurons activate the programmed cell death pathway and undergo apoptosis. Knowledge of how neurons undergo apoptosis is important for developing rational therapies that may prevent cell loss in several pathological situations such as stroke, spinal cord injury, and neurodegenerative diseases. We utilize various cell culture models, including cultures of sympathetic and cerebellar granule neurons to study the neuronal apoptotic pathway.

We are particularly interested in understanding how caspases become activated during neuronal apoptosis. The caspase proteases are critical executors of apoptosis; once activated, caspases induce rapid cell death. Caspase activation in most cells, including neurons, involves the release of cytochrome c from the mitochondria and the resulting formation of the apoptosome complex (comprising of Apaf-1 and procaspase-9). We have identified a novel pathway that regulates caspase activation in neurons and have recently focused our effort on the inhibitor of apoptosis (IAP) family of proteins as important components of this pathway. In addition to the ongoing research, we are interested in developing the following new projects:

1. Using immobilized IAP-binding peptides to identify novel IAPs from neuronal lysates
2. Using the Affymatrix technology to identify new genes that are induced in neurons undergoing apoptosis
3. Using RNAi technology to inhibit signaling pathways that control apoptosis
4. Investigating whether the apoptotic pathways that induce both soma (neuronal cell bodies) and neurite degeneration are identical
5. Constructing GFP fusions of apoptotic proteins to assess their movement within intracellular compartments during apoptosis with neuronal imaging studies

In addition to using the standard molecular and cell biological techniques (e.g. DNA manipulation, transfections, immunofluorescence, Western Blotting), the rotating students can learn how to isolate and culture neurons. They will also become familiar with various cell death assays and single-cell microinjections into neurons. Lastly, the students will be exposed to the latest literature on apoptosis through participation in our regular journal clubs.

For more information also visit our website at: http://deshmukhlab.web.unc.edu/#masthead