 | Gulden Kaplan The College of New Jersey BiologyMentor(s)Monica Driscoll Ph.D. Shaunak Kamat Department of Molecular Biology and Biochemistry Rutgers University |
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| Screening for roles of conserved Caenorhabditis elegans microRNAs in models of injury-induced neuronal necrosis | ||
| Micro-RNAs are small (~22nt) noncoding RNA molecules that bind to partially homolous sites on mRNA targets and suppress gene expression by degradation of the message or translational repression. miRNA regulation has many functional outcomes, including modulation of canonical pathways of cell death such as p53-mediated apoptosis. The Driscoll lab studies molecular mechanisms of necrotic neuronal death, with a major focus on toxicity of hyperactivated ion channels as occurs in stroke. Our goal is to test the influence of miRNAs in non-canonical death paradigms such as necrotic cell death induced by Ca2+ excitotoxicity, the underlying cause of neuronal damage in the event of trauma or seizure. A key initial aim is to normalize genetic backgrounds of C. elegans miRNA deletion strains by outcrossing with wild-type N2 nematodes. The outcrossed mutant strains will then be introduced in a mec-4(d) genetic background, which expresses a mutant hyperactive MEC-4 cation channel that remains constitutively open and causes necrosis by excessive Ca2+ influx into the cell. Necrosis is assayed in the six mechanosensory neurons of C. elegans by visualizing the Pmec-4::GFP reporter strain in impacted neurons under a standard epifluorescence microscope. Thus, miRNA deletion mutants miR-45(n4280) and miR-124(n4255), which are miRNAs that are conserved between nematodes and humans, may not only be necrosis suppressors, but also may identify a link between miRNAs and the necrotic cell death pathway. Our results should provide greater insight into the molecular mechanisms underlying the poorly understood process of neuronal necrosis and may hold implications for the treatment of neuronal injury following stroke, trauma or seizure. |