Dataset Information

NanoLC-MSMS TMT comparison of in vitro cultured/native reticulocytes and erythrocytes

ABSTRACT: Triplicate samples of donor-matched native reticulocytes (CD71+ cells), erythrocytes (CD71- cells) and in vitro culture-derived reticulocytes (derived from CD34+ cells) were analysed through TMT-based quantitative proteomics to examine possible protein abundance changes underlying reticulocyte maturation.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Erythrocyte, Blood Cell, Cell Culture, Reticulocyte

SUBMITTER: Pedro Moura

LAB HEAD: Ashley Mark Toye

PROVIDER: PXD009023 | Pride | 2018-08-03

REPOSITORIES: Pride

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Publications

Non-muscle myosin II drives vesicle loss during human reticulocyte maturation.

Moura Pedro L PL Hawley Bethan R BR Mankelow Tosti J TJ Griffiths Rebecca E RE Dobbe Johannes G G JGG Streekstra Geert J GJ Anstee David J DJ Satchwell Timothy J TJ Toye Ashley M AM

Haematologica 20180803 12

The process of maturation of reticulocytes into fully mature erythrocytes that occurs in the circulation is known to be characterized by a complex interplay between loss of cell surface area and volume, removal of remnant cell organelles and redundant proteins, and highly selective membrane and cytoskeletal remodeling. However, the mechanisms that underlie and drive these maturational processes <i>in vivo</i> are currently poorly understood and, at present, reticulocytes derived through <i>in vi ...[more]

PMID: 30076174

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Project description:The molecular interactions between the maternal environment and the developing embryo that are key for early pregnancy success are known to be influenced by factors such as maternal metabolic status. Our understanding of the mechanism(s) through which these individual nutritional stressors alter endometrial function and the in utero environment for early pregnancy success is, however, limited. Here we report, for the first time, the use of an endometrium-on-a-chip microfluidics approach to produce a multi-cellular endometrium in vitro, that was exposed to glucose and insulin concentrations associated with maternal metabolic stressors. Following isolation of endometrial cells (epithelial and stromal) from the uteri of non-pregnant cows in the early-luteal phase (Day 4-7 approximately), epithelial cells were seeded in the upper chamber of the device (4-6 104 cells/mL) and stromal cells seeded in the lower chamber (1.5-2 104 cells/mL). Three different concentration of glucose (0.5, 5.0 or 50 mM) or insulin (Vehicle, 1 or 10 ng/mL) were performed in the endometrial cells at a flow rate of 1µL/min for 72 hr to mimic the rate of secretion in vivo. Quantitative differences in the transcriptomic response of the cells and the secreted proteome of in vitro-derived uterine luminal fluid (ULF) were determined by RNA-sequencing and Tandem Mass Tagging Mass Spectrometry (TMT), respectively. Changes in maternal glucose altered 21 and 191 protein-coding genes in epithelial and stromal cells, respectively (p<0.05), with a dose-dependent quantitative change in the protein secretome (1 and 23 proteins in epithelial and stromal cells, respectively). Altering insulin concentrations resulted in limited transcriptional changes including transcripts for insulin-like binding proteins that were cell specific (5, 12, and 20) but altered the quantitative secretion of 196 proteins including those involved in extracellular matrix-receptor interaction and proteoglycan signaling in cancer. Collectively, these findings highlight one potential mechanism by which changes to maternal glucose and insulin associated with metabolic stress may alter uterine function.

Dataset Information

NanoLC-MSMS TMT comparison of in vitro cultured/native reticulocytes and erythrocytes

Publications

Non-muscle myosin II drives vesicle loss during human reticulocyte maturation.

Similar Datasets

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