Meeting Abstract
We utilize Lumbriculus variegatus, an aquatic oligochaete, to determine the effect of caffeine on the cellular and molecular mechanisms of regeneration. Utilizing bromodeoxyuridine (BrdU) labeling coupled with behavioral testing and segmentation, we determined the effects of short-term exposure to 2.5mM caffeine on epimorphic and morphallactic regeneration. Worms in both non-regenerating and regenerating populations were fed BrDU in an agarose and spirulina paste for one week before dissection to investigate stem cell migration. Regenerating populations of worms were then exposed to 2.5mM caffeine or left in Ozarka water as per Drewes, 1999. Total number of segments regenerated were determined over a three week period. Behavioral analysis was also carried out along the anterior to posterior body axis as evidence for morphallaxis and dissections were completed for stem cell migration analysis. Immunohistochemistry was performed on tissue samples with anti-BrDU and Lan 3-2 antibodies to track stem cell migration and upregulation of a glycoepitope marker of neural morphallaxis (Martinez et al., 2004). Preliminary data suggests that short-term exposure to 2.5mM caffeine reduced the number of head segments regenerated but did not appear to affect behavioral recovery along the anterior-posterior axis. Stem cell migration and Lan 3-2 expression patterns also appeared unchanged. Thus, caffeine exposure appears to reduce epimorphic regeneration but not morphallactic regeneration. With concern regarding the increased consumption of caffeine, Lumbriculus may prove to be valuable tool in fully understanding caffeine and its effects on a developing system.
