Dynamics of Zebrafish Heart Regeneration Using an HPLC–ESI–MS/MS Approach

Abstract

Failure to properly repair damaged due to myocardial infarction is a major cause of heart failure. In contrast with adult mammals, zebrafish hearts show remarkable regenerative capabilities after substantial damage. To characterize protein dynamics during heart regeneration, we employed an HPLC-ESI-MS/MS (mass spectrometry) approach. Myocardium tissues were taken from sham-operated fish and ventricle-resected sample at three different time points (2, 7, and 14 days); dynamics of protein expression were analyzed by an ion-current-based quantitative platform. More than 2000 protein groups were quantified in all 16 experiments. Two hundred and nine heart-regeneration-related protein groups were quantified and clustered into six time-course patterns. Functional analysis indicated that multiple molecular function and metabolic pathways were involved in heart regeneration. Interestingly, Ingenuity Pathway Analysis revealed that P53 signaling was inhibited during the heart regeneration, which was further verified by real-time quantitative polymerase chain reaction (Q-PCR). In summary, we applied systematic proteomics analysis on regenerating zebrafish heart, uncovered the dynamics of regenerative genes expression and regulatory pathways, and provided invaluable insight into design regenerative-based strategies in human hearts.

How to Stimulate Myocardial Regeneration in Adult Mammalian Heart: Existing Views and New Approaches

“…Accordingly, recent studies explored the potential of different approaches capable of stimulating cardiomyogenesis from mature CMs. For example, Ma et al [64] performed in-depth investigation of protein dynamics during heart regeneration of zebrafish using mass spectrometry. In comparison, Tahara et al [65] studied the potential effects of paracrine signals of epicardial and endocardial cells on cardiomyocyte proliferation in zebrafish, whereas Foglia and Poss [61] along with Galdos et al [66] attempted to approach the problem using heart development as a model.…”

“…For example, it was established that a link exists between the lack of telomerase activity and concomitant telomere dysfunction, which serve as natural barriers to cardiomyocyte proliferation and cardiac regeneration [67]. In addition, p53 signalling was inhibited during heart regeneration [64]. Furthermore, it was shown that the regeneration of damaged myocardium is mediated only by CMs that are adjacent to the injury area [63] whereas inflammation [68] and ROS promote regeneration [69].…”

The major vault protein is dispensable for zebrafish organ regeneration

“…Though an absolute function of cellular vaults has not been determined, several reports suggested these ribonucleoprotein particles might play a role in organ regeneration of zebrafish. Expression analysis revealed after injury, expression of MVP is induced in heart, caudal fin and spinal cord of zebrafish (Ma et al, 2018;Pan et al, 2013;Yoshinari et al, 2009). By using an morpholino-based knock-down approach, researchers observed reduced axonal regrowth after spinal cord transection and proposed MVP supports locomotor recovery and axonal regrowth in adult zebrafish (Pan et al, 2013).…”

A New Era of Cardiac Cell Therapy: Opportunities and Challenges

“…The heart of zebrafish, for example, was found to regenerate after serious injury, [7,8] due to the specificity of the myocardium environment and proliferative cardiomyocytes. [9] Additionally, neonatal mice cardiomyocytes were found to regenerate mainly through pre-existing cardiomyocytes. [4] Scientists now believe that the human heart is capable of some level of regeneration with varying degrees of myocardium renewal.…”