Publication Date

2008-09-30

Availability

Open access

Degree Type

Dissertation

Degree Name

Doctor of Philosophy (PHD)

Department

Neuroscience (Medicine)

Date of Defense

2008-05-22

First Committee Member

Abigail S. Hackam - Committee Chair

Second Committee Member

Robert W. Keane - Committee Member

Third Committee Member

John L. Bixby - Committee Member

Fourth Committee Member

Vance P. Lemmon - Mentor

Abstract

Several factors contribute to the failure of the central nervous system (CNS) to regenerate after injury. These include inhibition of axonal growth by myelin and glial scar associated molecules, as well as the intrinsic inability of adult CNS neurons to grow long axons in environments that are permissive for younger neurons. Neurons in the peripheral nervous system (PNS) display a much higher capacity to regenerate after injury than CNS neurons, as shown by conditioning lesion experiments and by microtransplantation of dorsal root ganglia neurons into CNS white matter tracts. Our central hypothesis is that neurons of the PNS express specific regeneration associated genes that mediate their enhanced growth response after injury. We have employed a combination of subtractive hybridization, microarray comparison and promoter analysis to probe for genes specific to neurons of the dorsal root ganglia (DRG), using cerebellar granule neurons (CGN) as a reference. We have identified over a thousand different genes, many of whose products form interaction networks and signaling pathways. Moreover, we have identified several dozen transcription factors that may play a role in establishing DRG neuron identity and shape their responses after injury. One of these transcription factors is Signal Transducer and Activator of Transcription 3 (STAT3), previously known to be upregulated in the PNS after a conditioning lesion but not known to be specific to the PNS. Using a real time PCR and immunochemical approaches we have shown that STAT3 is constitutively expressed and selectively active in DRG neurons both in culture and in vivo. We show that the overexpression of wild type STAT3 in cerebellar granule neurons leads to the formation of supernumerary neurites, whereas the overexpression of constitutively active STAT3-C leads to a 20% increase in total neurite outgrowth. It is hoped that the genetic delivery of STAT3-C, potentially combined with co-activators of transcription, will improve functional regeneration of CNS axons in vivo.

Keywords

Microarray; Subtraction; CNS; Axon Regeneration; Bioinformatics; Transcription; PNS; Dorsal Root Ganglia; DRG; Cerebellum

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