Meeting Abstract

P1.59  Jan. 4  A novel methodology for localized exposure of chorioallantoic membrane vessels to toxins affecting angiogenesis. KUMORDZIE, A*; BURRGREN, W; Univ. of North Texas; Univ. of North Texas

The chorioallantoic membrane (CAM) of the avian egg is a model vascular bed for understanding how toxins affect angiogenesis. Localized delivery of toxins is crucial to prevent indirect effects by heart rate (fH) alteration. We developed a technique for local delivery of alcohol (EtOH) to the CAM of Day 3 chicken embryo. EtOH is of particular interest because of its demonstrated negative effects on embryonic vessel growth. However, broad application of EtOH to the body and vasculature of the embryo is likely to have broad ramifications, hence our technique, applicable to the CAM of shell-less cultured chicken embryos. A delivery system was formed from a 2mm diameter cotton “wick” in the tip of a 1mL syringe. The syringe barrel was suspended through a plastic cover over the culture dish and adjusted to come into gentle, non-deforming contact with the CAM vessels of the growing embryo. A 100µL test solution was then placed into the barrel, wetting the wick in contact with the CAM remote from the embryo itself. Test solutions containing saline with 0.1% Evans Blue (sham) or EtOH (to final dilutions of 10%, 30%, 50%). The dye was essential to show the extent of spread of test solution from the wick. In these preliminary experiments, fH was monitored hourly over the first 24hr interval after initial exposure. EtOH did not significantly affect fH at any dose, despite migration of the solution for up to 10mm from the wick, as evidenced by spread of the dye. That EtOH, known to depress embryonic cardiac activity, did not change fH in our experiments indicates that its delivery was indeed localized to the CAM vessels. That is, EtOH did not spread systemically in concentrations high enough to depress fH, validating this technique for future examination of EtOH-angiogenesis interactions.