Botryllus schlosseri is a colonial chordate species that exhibits robust stem cell-mediated regeneration capacities throughout life. It grows through a process of colonial budding known as blastogenesis, in addition to its sexual reproduction mode, embryogenesis. In natural conditions, colony growth peaks in the summer and drastically decreases in the winter due to various seasonal factors. We have established and optimized a method to rear B. schlosseri animals in landlocked laboratories where the controlled conditions allow colonies to grow continuously and exponentially through asexual reproduction under constant temperature, salinity, light, and nutrient conditions. During the weekly blastogenic cycle, all asexually derived parental zooids synchronously regress within one day and are replaced by a new generation. To enable comprehensive molecular phenotyping of this species we established a quantitative proteomics workflow and spectral library for Liquid chromatography/Tandem mass spectrometry (LCMS) data-independent acquisition (DIA). In this study, we quantified the relative abundances of several thousand proteins representing molecular phenotypes of B. schlosseri. This workflow enables us to use proteomics to characterize zooid development during its regenerative cycle, including the short critical period of the takeover stage, during which degenerating zooids undergo massive apoptosis while succeeding buds quickly mature and migrate into place. From this data, protein networks associated with regeneration and degeneration can be created, offering insights into tissue developmental pathways in vivo and future cell immortalization strategies in vitro. Furthermore, the establishment of quantitative proteomics workflows for B. schlosseri coupled with its unique life cycle features promotes the use of this model organism for the study of phenotypic plasticity and evolution of aging, stem cells, and mechanisms of regeneration and cell differentiation. This project is funded by NSF Grant MCB — 2127516. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.