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Physiologic Hypertrophy

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Physiologic hypertrophy is the growth of cardiomyocytes due to stress. It is initiated by the binding of different ligands to receptors which result in the activation of PI3K-AKT and mTOR pathways. PI3K is a heterodimeric lipid kinase that is found in the plasma membrane and is activated once VEGFB binds to its VEGFR1 or IGF1 binding to IGF1R. Activated PI3K would cause for the activation of AKT. AKT is a serine/threonine protein kinase that would directly promote protein translation through the inhibition of GSK3β activity. GSK3β would negatively regulate eukaryotic translation initiation factors. AKT also inhibits the activity of FOXO3, thus resulting in reduced protein turnover and catabolism through ubiquitin ligases. As a result, this …show more content…
Conditional Groot KO mice will be exposed to tamoxifen shortly after birth to cause for gene KO. Conditional Groot overexpression mice will be exposed to tetracycline shortly after birth as well to increase levels of GROOT. To measure protein levels of physiological and pathological hypertrophy proteins, cardiomyocytes (CM) will be harvest from each cohort and will be grown in vitro. CM will then be lysed to be used in qt-PCR. Qt-PCR will quantify the expression levels of IGF1, ANP, and β-MHC. CM protein extracts will also be used in western blot analysis to detect phosphorylated products of both physiological (RHEB, S6K) and pathological (MAPKKK and MAPKK) hypertrophy. If our hypothesis is correct than in the KO mice, we should be seeing very minimal expression of IGF1 and no change in the expression of ANP, and β-MHC. We should also be seeing very little phosphorylated products of RHEB and S6K and no change in MAPKKK and MAPKK levels (as compared to WT and OE mice). The WT mice should be seeing a larger expression of IGF1 and again no change in the expression of ANP, and β-MHC (i.e. expression levels are the same with KO mice). WT mice should also have an increase amount of RHEB and S6K and no change in MAPKKK and MAPKK levels as compared to KO. Finally, the OE mice should have the largest levels of expression of IGF1 and again no …show more content…
One cohort of mice will undergo sham surgery while another cohort of mice will undergo TAC surgery to increase pressure on the heart. Hearts will be harvested during neonatal stages and will be placed under immunohistochemical analysis to analyze the width-to-length ratio of CM. Hearts will also be cut coronally to investigate septum thickness (organized growth). CM will also be harvested from the mice and qt-PCR and western blot analysis will be used to confirm results seen in experiment 1 and 2. KO mice will have very low expression of IGF1 and high expression of ANP and β-MHC and will have very low levels of RHEB and S6K and high levels of MAPKKK and MAPKK. These mice will also have not maintained the width-to-length ratio of CM and will have an extremely thick septum. These mice will also have very high levels of fibrosis and stiffness of the heart which are associated with pathological hypertrophy. WT mice will have high expression of IGF1 and low expression of ANP and β-MHC and will have high levels of RHEB and S6K and low levels of MAPKKK and MAPKK. These mice will have maintained the width-to-length ratio of CM and will have a similar septum size to sham mice. WT mice will also have very low levels of fibrosis and stiffness in the heart due to favourable remodeling through physiological hypertrophy. OE mice will have very high expression of IGF1 and low expression of ANP and β-MHC and will

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