The 2003 Annual Meeting of OASYS_NEW

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Differential Gene Expression in Human Peripheral Nerve and Neuromas

Siu HYH1, Mainprize TG2, Taylor MD2, Munro CA1, and Midha R1. (1) Division of Neurosurgery, Sunnybrook & Women's College HSC, University of Toronto, R106, 2075 Bayview Ave, Toronto, ON, Canada, (2) Brain Tumour Research Centre, Hospital for Sick Children, University of Toronto, Toronto, ON, Canada

Neuromas are the result of frustrated regeneration of axons in the PNS after injury to the nerve. Painful neuromas and neuromas in continuity that are not capable of successful regeneration are traditionally treated via surgical resection. The advent of microarray technology has allowed for large-scale simultaneous analysis of gene expression between two different tissue samples. The long-term goal of this project is to ascertain genes that are differentially expressed in human neuroma tissue. Such genes may include axonal guidance genes (eg: LAR and Shh), neuronal repair genes (eg: MAPK and ApoD), growth factors (eg: neurotrophins) and their receptors, and Schwann cell markers (eg: S100 and Krox20). In addition to these genes, novel genes are expected to be discovered as well. How these novel genes are involved in axonal regeneration, neuroma formation and pain generation has the potential to lead to future studies of specific genes.

METHODS: All tissue samples were collected at the time of surgery and immediately flash frozen in liquid nitrogen. Samples were then stored at –80oC until the RNA was extracted. RNA from human peripheral nerve (n=5) and neuroma tissue (n=8) was extracted via the TRIzol method. The RNA was amplified and fluorescently labelled via RT-PCR. After overnight hybridization to a 19K microarray chip, confocal laser microscopy and computer analysis yielded a list of differentially expressed genes.

RESULTS: Preliminary results show that there are differentially expressed genes between the two tissue types. However, due to the vast amount of information obtained, definitive results are not yet available. Ongoing analysis will allow precise determination of the differential expressed genes and gene families; this information will be available to present at the meeting.

NEXT STEPS: Immunohistochemistry will provide protein confirmation of the differentially expressed genes. Additional molecular analysis will facilitate the development of possible genetic hypotheses aimed at explaining neuroma formation and maintenance.


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