In vivo proximity labeling identifies cardiomyocyte protein networks during zebrafish heart regeneration

Abstract

Strategies have not been available until recently to uncover interacting protein networks specific to key cell types, their subcellular compartments, and their major regulators during complex in vivo events. Here we apply BioID2 proximity labeling to capture protein networks acting within cardiomyocytes during a key model of innate heart regeneration in zebrafish. Transgenic zebrafish expressing a promiscuous BirA2 localized to the entire myocardial cell or membrane compartment were generated, each identifying distinct proteomes in adult cardiomyocytes that became altered during regeneration. BioID2 profiling for interactors with ErbB2, a co-receptor for the cardiomyocyte mitogen Nrg1, implicated Rho A as a target of ErbB2 signaling in cardiomyocytes. Blockade of Rho A during heart regeneration, or during cardiogenic stimulation by the mitogenic influences Nrg1, Vegfaa or Vitamin D, disrupted muscle creation. Our findings reveal proximity labeling as a useful resource to interrogate cell proteomes and signaling networks during tissue regeneration in zebrafish.

Identification of the centrosomal maturation factor SSX2IP as a Wtip-binding partner by targeted proximity biotinylation

“…Pulldowns with neutravidin beads contained many additional bands in the sample from myc-BirA-WtipN-expressing embryos ( Fig 1E ), presumably corresponding to WtipN-interacting proteins. These results suggest that BirA can be used directly for proximity biotinylation in Xenopus , complementing previous observations for this and other models [ 25 – 28 ].…”

Turning gray selenium into a nanoaccelerator of tissue regeneration by PEG modification

“…Recently, zebrafish, a nonmammalian vertebrate, has become one of the top disease models for investigation because the similarity of its genome reaches as high as 70% with that of humans [ 33 ]. It can regenerate its heart [ 34 , 35 ], retina [ 36 ], kidney [ 37 , 38 ], liver [ 39 , 40 ], and appendants, for instance, the fins [ [41] , [42] , [43] ], throughout its entire life, which makes zebrafish one of the most popular models for studying tissue regeneration. Among regenerable organs, the tail fin, which is well organized with vessels, nerves, and stromal structures and comprises skin, mesenchyme, and bone frames, has become a popular model for studying tissue regeneration due to its repeatability, ease of manipulation, and timesaving [ 44 ].…”

The Role of Macrophages During Zebrafish Injury and Tissue Regeneration Under Infectious and Non-Infectious Conditions

“…These two types of regeneration, epimorphic and compensatory, are both required to regain a functional heart ( 24 ). Recently, the proliferation of cardiomyocytes during the regeneration of the zebrafish heart was shown to depend on myocardial Tp53 suppression by mitogen-induced Mdm2 and Rho activity ( 79 , 80 ).…”