Proteomic Analyses of the Developing Chicken Cardiovascular System

Up until today, no proteomics approaches have been described for heart muscle development. We describe a proteomics method to study the proteome of different heart structures at three stages of chicken embryonic development. For this purpose, a combination of gel separation, nanoLC separation and mass spectrometry was used. With this method, we identified in total 267 proteins in different tissue structures of chicken heart. We observed differences in protein abundance for a number of proteins between the different tissue structures and time points of development using spectral counting as a semiquantitative measure of protein abundance. For myosin-heavy chain 6, myosin-heavy chain 7, titin, connectin, collagen alpha-1, and xin, differences in protein levels for the different stages and structures (great arteries, outflow tract and ventricles) have been observed. A pathway analysis is performed in which the identified proteins are related to theoretical protein networks. Most prominent was the ‘cardiovascular system development and function’ network with the abundantly present proteins myosin 6 and myosin 7. We showed that myosin 6 is highly regulated in a stage and heart tissue specific manner. In conclusion, this method can be used to study changes in protein levels of chicken heart tissue in a spatiotemporal manner.