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Hiding Inside? Intracellular Expression of Non-Glycosylated C-Kit Protein in Cardiac Progenitor Cells

Hiding Inside? Intracellular Expression of Non-Glycosylated C-Kit Protein in Cardiac Progenitor Cells

Huilin Shi, Christopher A Drummond, Xiaoming Fan, Steven T Haller, Jiang Liu, Deepak Malhotra, Jiang Tian

Stem Cell Res. 2016 May;16(3):795-806.

PMID: 27161312

Abstract:

Cardiac progenitor cells including c-kit(+) cells and cardiosphere-derived cells (CDCs) play important roles in cardiac repair and regeneration. CDCs were reported to contain only small subpopulations of c-kit(+) cells and recent publications suggested that depletion of the c-kit(+) subpopulation of cells has no effect on regenerative properties of CDCs. However, our current study showed that the vast majority of CDCs from murine heart actually express c-kit, albeit, in an intracellular and non-glycosylated form. Immunostaining and flow cytometry showed that the fluorescent signal indicative of c-kit immunostaining significantly increased when cell membranes were permeabilized. Western blots further demonstrated that glycosylation of c-kit was increased during endothelial differentiation in a time dependent manner. Glycosylation inhibition by 1-deoxymannojirimycin hydrochloride (1-DMM) blocked c-kit glycosylation and reduced expression of endothelial cell markers such as Flk-1 and CD31 during differentiation. Pretreatment of these cells with a c-kit kinase inhibitor (imatinib mesylate) also attenuated Flk-1 and CD31 expression. These results suggest that c-kit glycosylation and its kinase activity are likely needed for these cells to differentiate into an endothelial lineage. In vivo, we found that intracellular c-kit expressing cells are located in the wall of cardiac blood vessels in mice subjected to myocardial infarction. In summary, our work demonstrated for the first time that c-kit is not only expressed in CDCs but may also directly participate in CDC differentiation into an endothelial lineage.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP73465437	1-Deoxymannojirimycin hydrochloride		73465-43-7	Price

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