Furthermore, we observed enhanced mortality in older transgenic dams, which may possibly be explained by multiple pregnancies related cardiac remodeling, exaggerating the existing cardiac hypertrophy. Transgenic mice from both lines exhibit marked cardiac hypertrophy as evident from gross pathology and radiographs. Heart excess weight tobody fat ratios have been comparable in transgenic and wild style mice as much as postnatal day six.five, with a substantial 54 improve at P9.5 . Heart bodyweight to tibia length ratio distinctions reached a highest of about a fold distinction at 10 12 weeks of age , right after which slight, age related parallel increases in the heart excess weight to tibia length ratio had been observed in the two transgenic and wild kind mice . Protein levels of ErbB2in hearts of transgenic mice have been measured by Western blot analysis , and comparable amounts of ErbB2 protein have been observed in both lines . Lysates through the left ventricles of 8 weeks outdated transgenic mice and wild style littermates had been compared.
The ErbB2 protein compound screening levels were around 40 instances greater in left ventricles of transgenic animals when compared with wild variety littermates, even though ErbB2 amounts in other organs were not altered . There have been no variations in protein expression involving male and female mice or between E3 and E6 lines, thus E6 males had been made use of for your vast majority of subsequent experiments. We evaluated ErbB2 protein expression in wild form and ErbB2 transgenic mice at days P and P6.five . Wild form mice have greater ErbB2 expression at P in comparison with wild variety mice at P6.5 days. In ErbB2 transgenic mice resulting from the transgene promoter activation, ErbB2 expression dramatically increases between P and P6.5 days. At P, heart weights are usually not various between wild sort and ErbB2 transgenic mice, but at P9.
5 the main difference gets to be vital. At P, ErbB2 protein expression in transgenic hearts is increased compared to wild form hearts prior to the hypertrophy gets to be evident, but the relative ErbB2 protein expression among wild form and ErbB2 transgenic mice is a great deal greater in P6.five and in adult animals compared to P mice reflective of the myosin informative post hefty chain promoter transgene activation. ErbB2 In excess of expression Triggers Activation of Prohypertrophic Gene System and Professional survival and Translational Pathways We confirmed by qRT PCR that grownup mice have elevated amounts of mRNA encoding for atrial natriuretic peptide and b myosin heavy chain , two characteristic mRNAs and proteins that raise in cardiac hypertrophy .
To investigate signaling pathways impacted by ErbB2 overexpression, we performed a series of experiments to assess ranges of proteins and amounts of protein phosphorylation for primary parts of the ErbB2 pathway. Left ventricles in the two lines of ErbB2 transgenic mice showed remarkably related patterns of protein expression or pathway activation.