Evidence for active acetylcholine metabolism in human amniotic epithelial cells: applicable to intracerebral allografting for neurologic disease.
Human amniotic epithelial (HAE) cells have been used for allotransplantation in patients with lysosomal storage disease due to lack of expression of HLA antigens. Previously, we have reported the expression of differentiation markers for both neural stem cells, and neuron and glial cells. In the present study, we investigated the presence of choline acetyltransferase (ChAT) and acetylcholine (ACh) in HAE cells using different experimental approaches. Cultured HAE cells showed strong immunoreactivity against ChAT antibody. ChAT activity in primary cells was 24.9 +/- 8.5 pmol/mg protein/h. Using HPLC with electrochemical detection, ACh was detected in both cell incubation media and cell pellets indicating that these cells synthesize and release ACh in a time-dependent manner. Additional confirmation of this hypothesis was gained from the data obtained from RT-PCR and Western blot analyses which revealed the expression of ChAT mRNA and ChAT protein, respectively, in HAE cells. Results of the present study suggest that HAE cells can possibly be applied for intracerebral allografting to treat neurologic diseases in which cholinergic neurons are damaged.
Sakuragawa
N
N
Department of Inherited Metabolic Disease, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Tokyo, Japan. sakuraga@ncnaxp.ncnp.go.jp
Misawa
H
H
Ohsugi
K
K
Kakishita
K
K
Ishii
T
T
Thangavel
R
R
Tohyama
J
J
Elwan
M
M
Yokoyama
Y
Y
Okuda
O
O
Arai
H
H
Ogino
I
I
Sato
K
K
D000109:Acetylcholine / Q000378:metabolism*; D000653:Amniotic Fluid / Q000166:cytology*; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D017690:Cell Transplantation / Q000379:methods*; D002795:Choline O-Acetyltransferase / Q000032:analysis / Q000235:genetics / Q000276:immunology; D002799:Cholinergic Fibers / Q000201:enzymology; D004847:Epithelial Cells; D004848:Epithelium / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007150:Immunohistochemistry; D009422:Nervous System Diseases / Q000628:therapy*; D010920:Placenta / Q000166:cytology; D012333:RNA, Messenger / Q000032:analysis; D014184:Transplantation, Homologous
D000911:Antibodies, Monoclonal; D012333:RNA, Messenger; D002795:Choline O-Acetyltransferase; D000109:Acetylcholine