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

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.