2005 Fiscal Year Final Research Report Summary
Establishment and analysis aria vitro model neurons lifaLEVEed from transgenic ES cells, in which a FALS-linked mutant SOD1 is introduced
| Project/Area Number |
16590845
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Neurology
|
|---|
| Research Institution | Keio University |
|---|
Principal Investigator |
ABE Yokbiro Keio University, School of Medicine, Instructor, 医学部, 助手 (10317331)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
NIIKURA Takaio Keio University School of Meclecine, Instructor, 医学部, 助手 (10301491)
|
|---|
| Project Period (FY) |
2004 – 2005
|
| Keywords | embryonic stem cell / amyotrophic lateral sclerosis / disease model / differentiation / spinal motor neurons / SOD1 / ALS |
|---|
| Research Abstract |
An inducible transgene that expresses FALS-linked mutant SOD1 was introduced into both NSC34 cells and mouse embryonic stem (ES) cells and several stable clones were obtained. Cre recombinase was transiently expressed using adenovirus vector in both the transgenic NSC 34 cells or neurons derived from the transgenic ES cells to induce the mutant SOD1 in these cells. However, we were unable to detect any significant cell death in these cells.
To establish more specific disease model, we decided to differentiate mouse ES cells into spinal motor neurons, which is one of the most vulnerable neurons in ALS. For this purpose, we developed three types of stable CHO-cell lines expressing Sonic hedgehog (Shh) with distinct lipid modification(s) (N-terminal palmitoylation and C-terminal chlesterylation). We are now trying to combine these Shh-producing cells with our original protocol for differentiation of ES cells into neuronal cells.
In addition, using these Shh-producing CHO-cell lines and transient transfection system of several mammalian cells, we examined intracellular trafficking and secretion of Shh from the cells and found that the palmitoylated Shh are hardly secreted probably because they are highly accumulated in some intracellular compartment, especially in ER. Extracellular carrier was also necessary for Shh to be secreted efficiently. We also found a negative regulator for palmitoylation of Shh.
|
Research Products
(2 results)
