Effective Rapid Blood Perfusion inXenopus

Abstract

ABSTRACTXenopushas been a powerful model organism for understanding vertebrate development and disease for over a hundred years. Here we define a rapid blood perfusion protocol inXenopusaimed at a consistent and drastic reduction of blood across tissues. Perfusion is done by inserting a needle directly into the ventricle and pumping heparin in PBS through the vascular system. The whole procedure should take about 10 minutes per frog. Blood is dominated by a few highly abundant proteins and cell types which create numerous issues by masking most other molecules and cell types of interest. Reproducible characterization of adultXenopustissues with quantitative proteomics and single cell transcriptomics will gain from applying this protocol prior to organ dissections defined in companion papers. The procedure is aimed at standardization of practice across the animals of different gender, age andXenopusspecies, specificallyX.laevisandX.tropicalis.SUMMARYAn effective rapid blood perfusion protocol to prepare tissue samples for transcriptomics and proteomics studies.