2000 Fiscal Year Final Research Report Summary
Genetic introduction of thyroid hormone transporter in Bood-Brain-Barrier
| Project/Area Number |
11557188
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B).
|
| Allocation Type | Single-year Grants |
|---|
| Section | 展開研究 |
|---|
| Research Field |
医薬分子機能学
|
|---|
| Research Institution | Tohoku University |
|---|
Principal Investigator |
ABE Takaaki Tohoku Univ., School of Med., Neurophysiology, Research Associate, 大学院・医学系研究科, 助手 (80292209)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
YAWO Hiromu Tohoku Univ., School of Med., Neurophysiology, Porfessor, 大学院・医学系研究科, 教授 (00144353)
|
|---|
| Project Period (FY) |
1999 – 2000
|
| Keywords | organic anion transporter / Thyroid hormone transporter / adenovirus / blood-brain-barrier / choroid plexus / gene transfer |
|---|
| Research Abstract |
We have established adenovirus-mediated gene transfer system into cells.
The coding region of LST-1 and LST-2, both of which have isolated by us have introduced into adenovirus vector, packaged, and transfected into HEK293 cell with adenovirus genomic DNA.The cells that produces homologous-recombinant shrieked and made plaque formation. After producing large scale infected medium, the recombinant adenovirus was purified by CsCl ultracentrifuge.
The infection titer of the purified LST-1 and LST-2 adenoviruses were measured and the IC 50 of each virus was 1.26 and 1.48 X 10^<10> pfu, respectively.
When recombinant virus was applied to cultured cell line, the LST-1 and LST-2 protein was strongly expressed within 24 hr after application. The size of the band was also confirmed by Western blot analysis. The LST-1 and LST-2 expressing mammalian cell transports various organic anions as substrates and the apparent Km value of tauro-cholate uptake is 24 micro M and 32 micro M, respectively and both of which is a comparative value of the native cells reported.
The establishment of the adenovirus-mediated gene transfer system wiil lead to the various applied sysem for screening of the brain-, liver and kidney oriented drug delivery and therapeutic tools.
|
Research Products
(12 results)
-
-
-
-
-
-
-
[Publications] Abe T., Kakyo M., Tokui T., Nakagomi R., Nishio T., Nakai D., Nomura H., Unno M., Suzuki M., Naitoh T., Matsuno S.and Yawo H.: "Identification of a novel gene family encoding human liver-specific organic anion transporter LST-1."J.Biol.Chem.. 274. 17159-17183 (1999)
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Abe T., Kakyo M., Sakagami H., Tokui T., Nishio T., Tanemoto M., Nomura H., Hebert S.C., Matsuno S., Kondo H., and Yawo H.: "Molecular characterization and tissue distribution of a new organic anion transporter subtype (oatp3) that transport thyroid hormones and taurocholate and comparison with oatp2."J.Biol.Chem.. 273. 22395-22401 (1998)
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Kakyo M., Unno M., Tokui T., Nakagomi R., Nishio T., Iwasashi H., Nakai D., Seki M., Suzuki M., Naitoh T., Matsuno S., Yawo H., and Abe T.: "Molecular characterization and functional regulation of a novel rat liver-specific organic anion transporter rlst-1"Gastroenterology. 117. 770-775 (1999)
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Kakyo M., Sakagami H., Nishio T., Nakai D., Nakagomi R., Tokui T., Naitoh T., Matsuno S., Kondo H., Abe T., and Yawo H.: "Immunohistochemical distribution and functional characterization of an organic anion transporting polypeptide2 (oatp2)."FEBS Lett.. 445. 343-346 (1999)
Description
「研究成果報告書概要(欧文)」より
-
-
[Publications] Nishio T., AdachiH., NakagomiR., OnogawaT., Fujiwara K., Sato E., Tokui T., Unno M., KakyoM., Nakai D., Tanemoto M., Okabe M., SuzukiT., Suzuki M., Ohtani H., Kondo Y., Ito S., Matsuno S., Iinuma K.Yawo H.and Abe T.: "Molecular identification of a novel organic anion transporter moatpl, which transports prostaglandin D2, leukotriene C4 and taurocholate."Biochem.Biophys.Res.Commun.. 275. 946-954 (2000)
Description
「研究成果報告書概要(欧文)」より
