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Atrial Natriuretic Peptide Regulates Calcium Approach in Mouse ES Derived Cardiomyocytes

Author: ChenSuHua
Tutor: LiYunLong；JiGuangJu
School: Shandong Normal University
Course: Cell Biology
Keywords: Atrial natriuretic peptide Embryonic stem cells Differentiation Calcium current Action potential
CLC: R33
Type: Master's thesis
Year: 2007
Downloads: 71
Quote: 0
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Abstract


Purpose : L - type calcium channels are voltage-gated ion channels involved in the myocardial extracellular calcium flow in the plateau phase of the action potential , and play an important role in myocardial contraction coupling process . Atrial natriuretic peptide is a peptide secreted by the heart , which can cut the L-type calcium current , but the mechanism of action is unclear . This article aims to explore the regulatory mechanism of the L-type calcium current in cardiomyocytes of atrial natriuretic peptide on mouse embryonic stem cell differentiation . Methods : application of embryonic stem cell hanging drop culture induced to differentiate into cardiomyocytes , the use of collagenase II digestion for different periods of single beating cardiomyocytes ; using whole-cell patch clamp technique to record the L-type of atrial natriuretic peptide stimulation of myocardial cells calcium current and signal channel inhibitor or activator of L-type calcium current regulation . Immunofluorescence assay measured cell -specific protein ANP and myosin MHC expression ; detect the expression of ANP levels in different periods of mouse embryonic heart . Results : Using cultured mouse embryonic stem cells and differentiate into beating EB hanging drop method , the enzyme digestion to obtain single beating cardiomyocytes , the immunofluorescence detected cardiac specific а-MHC expression , and is detected in the early ESMC cardiomyocytes action potential and calcium current . ANP in EDS and LDS myocardial cells in action potential significant negative chronotropic effect and can reduce the basis of the EDS and LDS cardiomyocytes and Iso activate L-type calcium current . ANP reduce ILcal pathway independent of the NO pathway , PTX - sensitive G-protein (Go, Gi) and soluble the GC . ANP inhibition of ILcal through its receptor activates particulate GC , cGMP -dependent PDE II pathway PDE II hydrolyse cAMP lead concentration decreased , reducing the regulation of calcium channel opening frequency . RT-PCR results showed that ANP expression during embryonic development continues to rise , and in eight days the embryo has already expressed , 12 days ANP strongly expressed the subsequent expression of ANP basically did not change , but after birth increased secretion and expression of adult reduced. The immunofluorescence detection ANP positioning dispersed in the cytoplasm in embryonic expression , the early cellular ANP expression appears polarity .

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