Effects of daunorubicin on ganglioside expression and neuronal differentiation of mouse embryonic stem cells.
Gangliosides are implicated in neuronal development processes. The regulation of ganglioside levels is closely related to the induction of neuronal cell differentiation. In this study, the relationship between ganglioside expression and neuronal cell development was investigated using an in vitro model of neural differentiation from mouse embryonic stem (mES) cells. Daunorubicin (DNR) was applied to induce the expression of gangliosides in embryoid body (EB) (4+). We observed an increase in expression of gangliosides in all stages of EBs by treatment of DNR (2microM). High-performance thin-layer chromatography (HPTLC) showed that gangliosides GD3, GD1a, GT1b, and GQ1b increased in DNR-treated 7-day-old EB (4+) [EB (4+):7]. DNR treatment significantly increased the expression of gangliosides, especially GT1b and GQ1b in comparison to control cells. Interestingly, GQ1b co-localized with microtubule-associated protein 2 (MAP-2) expressing cells in DNR-treated EB (4+):7. The co-localization of GQ1b and MAP-2 was found in neurite-bearing cells in DNR-treated 15-day-old EB (4+) [EB (4+):15], whereas no significant expression of GQ1b and less neurite formation were observed in untreated control. Also, the expression of synaptophysin and NF200, both neuronal markers associated with neruites, was increased by DNR treatment. These results demonstrate that DNR increases expression of gangliosides, especially GQ1b, in differentiating neuronal cells. Further, neurite-bearing neuronal cell differentiation can be facilitated by DNR, possibly through the induction of gangliosides. Thus, the present data suggest that DNR is beneficial for facilitating neuronal differentiation from ES cells and among the gangliosides analyzed in the present study, GQ1b is mainly involved in neurite formation.
Lee
Dae-Hoon
DH
Department of Biological Science, College of Natural Sciences, Wonkwang University, 344-2 Shinyong-dong, Iksan, Jeonbuk 570-749, Republic of Korea.
Koo
Deog-Bon
DB
Ko
Kinarm
K
Ko
Kisung
K
Kim
Sun-Mi
SM
Jung
Ji-Ung
JU
Ryu
Jae-Sung
JS
Jin
Jung-Woo
JW
Yang
Hyo-Jung
HJ
Do
Su-Il
SI
Jung
Kyu-Yong
KY
Choo
Young-Kug
YK
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003630:Daunorubicin / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005732:Gangliosides / Q000096:biosynthesis*; D007150:Immunohistochemistry; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D013997:Time Factors
D000903:Antibiotics, Antineoplastic; D005732:Gangliosides; C021298:ganglioside, GD1a; C016481:trisialoganglioside GT1; C026226:ganglioside, GD3; C039129:GQ1b ganglioside; D003630:Daunorubicin