Invited Symposium: Molecular Mechanisms of Ageing
Discussion and Conclusion
Summary and Conclusions
1. DDRT-PCR screening of 10-12% of brain mRNA resulted in the isolation of 60 bands that were selectively enriched in synaptoneurosome RNA. Thus, an upper limit of 600 mRNAs representing 1-5% of the total population of brain mRNAs may be available for translation by synaptic polyribosomes.
2. Three of the twenty DDRT-PCR bands that were reamplified, subcloned and sequenced were found to be selectively enriched in all three preparations of synpatoneurosomes tested; these are Ax-actin, ceramide glucosyl transferase and a zinc binding protein, ZBP89, while others, such as calmodulin bind protein a protein known to be at synapses, were enriched in two of the three preparations. Nevertheless, not all of the RNAs were found to be selectively enriched in synaptoneurosomal RNA, showing that the procedure has a degree of batch-to-batch variability that requires confirmation by RT-PCR or Northern blotting using several preparations of synaptoneurosomes.
3. Having established this system, we can now begin to delineate the effects of development, learning and ageing on the synaptoneurosome-specific mRNAs we have isolated; we will also use it to expand the library of known synaptoneurosome enriched mRNAs.
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|Sung, Y-J.; Weiler, I. J.; Greenough, W. T.; Denman, R. B.; (1998). Selectively Enriched Messenger RNAs in Rat Synaptoneurosomes. Presented at INABIS '98 - 5th Internet World Congress on Biomedical Sciences at McMaster University, Canada, Dec 7-16th. Invited Symposium. Available at URL http://www.mcmaster.ca/inabis98/higuchi/sung0691/index.html|
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