Dataset Information

The N-cadherin interactome in primary cardiomyocytes as defined by quantitative proximity proteomics

ABSTRACT: Low levels of Cdh2-BioID2 were expressed in mouse primary neonatal cardiomyocytes using an adenoviral expression system. Infected cells were treated with biotin for 24 hours before lysis. Biotinylated proteins were precipitated from lysates using streptavidin beads. Protein samples were in-gel trypsin digested and the resulting tryptic peptides were analyzed on an LTQ Orbitrap Velos.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mus Musculus (ncbitaxon:10090)

SUBMITTER: Adam V. Kwiatkowski

PROVIDER: MSV000083362 | MassIVE | Thu Jan 24 18:34:00 GMT 2019

SECONDARY ACCESSION(S): PXD012496

REPOSITORIES: MassIVE

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Publications

Fascial space priority approach in laparoscopy: lateral pelvic lymph node dissection for advanced low rectal cancer.

Sun Y Y Zhang Z Z Zhou Y Y Zhang X X

Techniques in coloproctology 20200227 4

PMID: 32107685

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Project description:Cardiomyocytes exhibit differential growth patterns throughout development. In fetal life the increase in cardiac mass is associated with hyperplastic growth and cardiomyocyte proliferation. The majority of fetal cardiomyocytes are also mononucleated. During the early neonatal period in mice there is a switch from hyperplastic to hypertrophic growth of cardiomyocytes. This period is characterized by bi-nucleation and polyploidization of cardiomyocyte nuclei and a decreased capacity for cardiomyocytes to proliferate and complete cytokinesis. Increases in myocardial mass occur predominantly via hypertrophic growth. Adult mammalian cardiomyocytes are generally accepted to have little or no proliferative capacity and to be terminally withdrawn from the cell cycle. The vast majority of adult murine cardiomyocytes are bi-nucleated. The present study sought to accurately establish the growth pattern of cardiomyocytes throughout development in mice and identify genes associated with the switch from hyperplastic to hypertrophic growth. These cell cycle associated genes are crucial to the understanding of the mechanisms of bi-nucleation, polyploidization and hypertrophy in the neonatal period. Cardiomyocytes were FACS sorted from the hearts of ED11-12 embryos, neonatal day 3-4 and adult (10 week) eGFP ?-MHC mice whereby GFP expression is driven constitutively by the ?-MHC promoter. Gene analyses identified genes whose expression was predicted to be particular to day 3 -4 neonatal cardiomyocytes, compared to embryonic or adult cells. Cell cycle associated genes are crucial to the understanding of the mechanisms of bi-nucleation and hypertrophy in the neonatal period, and offer attractive candidates for manipulation. Total RNA obtained from isolated cardiomyocytes from ED11-12; Neonatal day 3-4 and adult timepoints compared with each other. Several hearts per sample, RNA was pooled within samples. FACS samples were prepared in the following manner: embryonic, neonatal and adult hearts were dissected and dissociated to single cell solution with Liberase Blendzyme 3 (0.1 mg/ml) (Roche Diagnostics), washed and spun down and resuspended in cardiomyocyte isolation buffer (130 mM NaCl; 5 mM KCl; 1.2 mM KH2PO4; 6 mM HEPES; 5 mM NaHCO3; 1 mM MgCl2; 5 mM Glucose).