Proteomic analysis of temperature-dependent developmental plasticity within the ventricle of juvenile Atlantic salmon (Salmo salar).

In teleosts, cardiac plasticity plays a central role in mediating thermal acclimation. Previously, we demonstrated that exposure to elevated temperatures throughout development (+4°C) improved acute thermal tolerance of the heart in juvenile Atlantic salmon. Fish raised in a warmer thermal regime also displayed higher proportions of compact myocardium within their ventricles. In the present study, we investigated the molecular mechanisms supporting this temperature-specific phenotype by comparing relative protein abundance in ventricular tissue from the same experimental fish using mass spectrometry. We provide the first description of the ventricular proteome in juvenile Atlantic salmon and identify 79 proteins displaying differential abundance between developmental treatments. The subset of proteins showing higher abundance in fish raised under elevated temperatures was significantly enriched for processes related to ventricular tissue morphogenesis, and changes in protein abundance support a hypertrophic model of compact myocardium growth. Proteins associated with the vasculature and angiogenesis also showed higher abundance in the warm-developmental group, suggesting capillarization of the compact myocardium in the hearts of these fish. Proteins related to oxidative metabolism and protein homeostasis also displayed substantive shifts in abundance between developmental treatments, underscoring the importance of these processes in mediating thermal plasticity of cardiac function. While rapid growth under warm developmental temperatures has been linked to cardiomyopathies in farmed salmon, markers of cardiac pathology were not implicated in the present study. Thus, our findings offer a molecular footprint for adaptive temperature-dependent plasticity within the ventricle of a juvenile salmonid.

Modulation of RECK levels in Xenopus A6 cells: effects on MT1-MMP, MMP-2 and pERK levels.

BACKGROUND: MT1-MMP is a cell-surface enzyme whose regulation of pro-MMP-2 and ERK activation position it as a key facilitator of ECM remodelling and cell migration. These processes are modulated by endogenous MMP inhibitors, such as RECK, a GPI-anchored protein which has been shown to inhibit both MT1-MMP and MMP-2 activity. Our previous studies have revealed a link between MT1-MMP levels, and pro-MMP-2 and ERK activation in mammalian cells, as well as MT1-MMP and RECK co-localization in Xenopus embryos. We here investigated how modulation of RECK would impact MT1-MMP and MMP-2 levels, as well as ERK signalling in Xenopus A6 cells. RESULTS: We used a Morpholino approach to knockdown RECK, plasmid transfection to overexpress RECK, and PI-PLC treatment to shed RECK from the cell surface of Xenopus A6 cells. RECK reduction did not alter pERK or MT1-MMP levels, nor MMP-2 activity as measured by zymography; thus RECK-knockdown cells maintained the ability to remodel the ECM. RECK overexpression and PI-PLC treatment both increased ECM remodelling potential through increased MT1-MMP protein and relative MMP-2 activation levels. CONCLUSIONS: RECK changes that reduce the ability of the cell to remodel the ECM (overexpression and cell surface shedding) are compensated for by increases in MT1-MMP, and MMP-2 levels as seen by zymography.