KMID : 0620920090410120935
|
|
Experimental & Molecular Medicine 2009 Volume.41 No. 12 p.935 ~ p.945
|
|
The roles of glycosphingolipids in the proliferation and neural differentiation of mouse embryonic stem cells
|
|
Jung Ji-Ung
Ko Ki-Sung Choo Young-Kug Ko Ki-Narm Chang Kyu-Tae Lee Dae-Hoon
|
|
Abstract
|
|
|
Glycosphingolipids including gangliosides play important regulatory roles in cell proliferation and differentiation. UDP-glucose:ceramide glucosyltransferase (Ugcg) catalyze the initial step in glycosphingolipids biosynthesis pathway. In this study, Ugcg expression was reduced to approximately 80% by short hairpin RNAs (shRNAs) to evaluate the roles of glycosphingolipids in proliferation and neural differentiation of mouse embryonic stem cells (mESCs). HPTLC/immunofluorescence analyses of shRNAtransfected mESCs revealed that treatment with Ugcg-shRNA decreased expression of major gangliosides, GM3 and GD3. Furthermore, MTT and Western blot/immunofluorescence analyses demonstrated that inhibition of the Ugcg expression in mESCs resulted in decrease of cell proliferation (P £¼ 0.05) and decrease of activation of the ERK1/2 (P £¼ 0.05), respectively. To further investigate the role of glycosphingolipids in neural differentiation, the embryoid bodies formed from Ugcg-shRNA transfected mESCs were differentiated into neural cells by treatment with retinoic acid. We found that inhibition of Ugcg expression did not affect embryoid body (EB) differentiation, as judged by morphological comparison and expression of early neural precursor cell marker, nestin, in differentiated EBs. However, RT-PCR/immunofluorescence analyses showed that expression of microtubule- associated protein 2 (MAP-2) for neurons and glial fibrillary acidic protein (GFAP) for glial cells was decreased in neural cells differentiated from the shRNA-transfected mESCs. These results suggest that glycosphingolipids are involved in the proliferation of mESCs through ERK1/2 activation, and that glycosphingolipids play roles in differentiation of neural precursor cells derived from mESCs.
|
|
KEYWORD
|
|
cell differentiation, ceramide glucosyltransferase, extracellular signal-regulated MAP kinases, glycosphingolipids, embryonic stem cells, neurons
|
|
FullTexts / Linksout information
|
|
|
|
Listed journal information
|
|
|
|