Repository logo
 

Effect of MicroRNA-34c on Regenerative Axon Growth in Dorsal Root Ganglion Neurons of the Peripheral Nervous System

dc.contributor.advisorMurashov, Alexander K.en_US
dc.contributor.authorRaafat, Mohamed Hussienen_US
dc.contributor.departmentBiologyen_US
dc.date.accessioned2012-05-20T15:20:31Z
dc.date.available2013-07-01T11:15:48Z
dc.date.issued2011en_US
dc.description.abstractThe peripheral nervous system is unique in its ability to regenerate axons after injury. Our lab previously assessed the miRNA expression levels of the regenerating sciatic nerve which indicated miR-34c up-regulation. MicroRNAs are non-protein coding small RNA molecules, ~20 nt in length, that have been shown to control many basic cellular processes by post-transcriptional regulation. The purpose of this study was to identify the effect of miR-34c on regenerating axons of DRG neurons. 9 to 12 week CD-1 male mice used for this study were subjected to sciatic nerve crush lesion. Five days after surgery, animals were sacrificed, and DRGs were dissected and dissociated physically and enzymatically. DRG neurons were then grouped as follows: 1) non-transfected, 2) miR-34c mimic, 3) miR-34c inhibitor, 4) mimic negative control, and 5) inhibitor negative control. Cells were grown for 48 hours on glass cover slips, fixed, and incubated with primary beta-tubulin III antibody and with secondary antibody conjugated with a fluorophore. Cells were viewed on an inverted fluorescent microscope, and images were captured using a digital camera. Axon length and branches were measured using ImageJ. Inhibition of miR-34c showed a 27% increase in axon length for naive cultures (p<0.05), and a 23% increase in regenerating cultures (p<0.05), when compared to the inhibitor negative control. Also, a potential miR-34c target named Neuron Navigator 1 (NAV1) expression showed 43% increase after inhibition (p<0.05). Over-expression of miR-34c showed a 16% decrease in branching of regenerating cultures (p<0.05), as well as a 52% decrease in NAV1 expression. In conclusion, miR-34c inhibitor promoted axon growth in regenerating cultures resulting in longer axons and increased NAV1 expression. Over-expression of miR-34c produced a degenerative response in regenerating neurons resulting in decreased branching and decreased NAV1 expression.  en_US
dc.description.degreeM.S.en_US
dc.format.extent82 p.en_US
dc.format.mediumdissertations, academicen_US
dc.identifier.urihttp://hdl.handle.net/10342/3834
dc.language.isoen_US
dc.publisherEast Carolina Universityen_US
dc.subjectPhysiologyen_US
dc.subjectBioinformaticsen_US
dc.subjectGeneticsen_US
dc.subjectAxonsen_US
dc.subjectDrgen_US
dc.subjectMicroRNA-34cen_US
dc.subjectMir-34cen_US
dc.subjectNerve regenerationen_US
dc.subjectPeripheral nervous systemen_US
dc.subjectBiology, Genetics
dc.subjectBiology, Physiology
dc.subject.lcshNerves, Peripheral--Wounds and injuries
dc.subject.lcshNervous system--Regeneration
dc.subject.lcshAxons
dc.subject.lcshRNA
dc.subject.lcshNeurogenetics
dc.titleEffect of MicroRNA-34c on Regenerative Axon Growth in Dorsal Root Ganglion Neurons of the Peripheral Nervous Systemen_US
dc.typeMaster's Thesisen_US

Files

Original bundle

Now showing 1 - 2 of 2
Loading...
Thumbnail Image
Name:
Raafat_ecu_0600M_10583.pdf
Size:
4.77 MB
Format:
Adobe Portable Document Format

Please login to access this content.

Download
Loading...
Thumbnail Image
Name:
MicroRNa-biomachinerypaper.pdf
Size:
2.3 MB
Format:
Adobe Portable Document Format

Please login to access this content.

Download