TY - JOUR
T1 - Evidence for active acetylcholine metabolism in human amniotic epithelial cells
T2 - Applicable to intracerebral allografting for neurologic disease
AU - Sakuragawa, Norio
AU - Misawa, Hidemi
AU - Ohsugi, Keiko
AU - Kakishita, Kouji
AU - Ishii, Takashi
AU - Thangavel, Ramasamy
AU - Tohyama, Jun
AU - Elwan, Mohamed
AU - Yokoyama, Yasunobu
AU - Okuda, Osamu
AU - Arai, Hajime
AU - Ogino, Ikuko
AU - Sato, Kiyoshi
N1 - Funding Information:
The authors thank Dr. M. Yamamoto (Shrive Center) and Dr. R. McKay (NIH) for their respective gifts of antibodies, RC1 and nestin, respectively. Dr. Y. Takashima (NCNP) kindly offered the brain tissue as a positive control for RT-PCR. This work is supported in part by a Research Grant for Highly Advanced Medical Treatment, a Grant from Japan Foundation for Neuroscience and Mental Health and a Science Grant (5A-6) for Nervous and Mental Disorders from the Ministry of Health and Welfare.
PY - 1997/8/22
Y1 - 1997/8/22
N2 - 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.
AB - 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.
KW - Acetylcholine
KW - Choline acetyltransferase
KW - Human amniotic epithelial cells
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U2 - 10.1016/S0304-3940(97)00570-3
DO - 10.1016/S0304-3940(97)00570-3
M3 - Article
C2 - 9292890
AN - SCOPUS:0030764188
SN - 0304-3940
VL - 232
SP - 53
EP - 56
JO - Neuroscience Letters
JF - Neuroscience Letters
IS - 1
ER -
