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The Roles of Telomerase in Regeneration during Aging and the Telomeric DNA Sequence Identification in Aeolosoma viride

Telomeres are the functional chromosome elements that protect the ends of eukaryotic linear chromosomes from degradation and fusion. Incomplete telomere DNA replication results in length shortening during every round of cell division. On the other hand, the enzyme telomerase can extend the ends of telomeres and prevent this situation to happen in certain tissues. With essential function for genome integrity and cell survival, it has been reported that short telomeres and defective telomerase activity are involved in several age‐related diseases. Moreover, recent studies suggest that telomerase may be important for tissue renewal and regeneration after injury. Aeolosoma viride is a kind of fresh water annelid. It has strong ability in regeneration and is easy to manipulate for experiments. After anterior four segments decapitation, A. viride can regenerate its lost parts within five days. However, the regeneration successful rate decreases when the animal gets older and older. The regulation of telomerase gene expression in regeneration and how aging affecting this process are not well understood, so we would like to explore this mysterious aspect. In this study, we cloned the gene of telomerase protein component, avi‐tert, from our A. viride transcriptome database. The results of in situ hybridization and quantitative PCR showed that avi‐tert would express at the regenerating site, the blastema, where cell proliferation is undergoing. Besides, to identify the telomere repeat sequence, we searched the telomerase database (http://telomerase.asu.edu/) and chose two possible sequences, which are TTAGGG and TTAGG, according to the phylogenetic relationship of our species. Then, we extracted the genomic DNA for dot blotting assay and terminal restriction fragment (TRF) analysis. The results suggested that the telomere sequence of A. viride is most possible to be “TTAGGG” (same as human's) rather than “TTAGG” (in some invertebrates). In the next step, we will measure the gene expression profile in the animals of different ages and try to determine whether there are any changes during regeneration process.