Abstract

Introduction

Materials & Methods

Results

Discussion & Conclusion

References

Discussion
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Local translation at synapses has been implicated in neuronal plasticity, learning and memory formation. Conversely, deficits in synaptic translation may be involved in ageing. We have used DDRT-PCR to identify the subset of mRNAs that are selectively enriched at synapses. Comparisons were made between rat cortex RNA and RNA preparations obtained from rat synaptoneurosomes. To date, approximately 10% of rat mRNA has been screened and 60 synaptoneurosome-enriched bands identified. Twenty of these bands were cloned and sequenced. Messenger RNAs required for translation (L18a & S29 ribosomal proteins), cell structure (AX actin), cell signalling (calmodulin vesicle-associated protein kinase), lipid modification (ceramide glucosyltransferase) and cell shape and remodelling (osteonectin) were among those found in the initial DDRT-PCR pool. Interestingly, a mRNA with homology to a zinc finger binding protein (ZBP-89) was present. Finally, two previously unidentified messages and four expressed sequence tags (ESTs) were also found. RT-PCR with primers designed to the coding sequence (CDS) of seven of the known mRNAs showed that Ax-actin, ZBP-89, and ceramide glucosyltransferase were selectively enriched in 3 synaptoneurosome preparations. Nested primers designed to three of the ESTs revealed that two of these were also differentially expressed in two synaptoneurosome preparations. These data begin to delineate the local pool of mRNAs that are available to respond to modulatory events at synapses. This work was supported by the FRAXA Foundation.

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Presentation Number SAsung0691
Keywords: ddrt-pcr, synaptoneurosomes, mRNA