Designation and evaluation of antisense oligodeoxynucleotides targeted to glial glutamate transporter-1a.
- Author:
Li-zhe LIU
;
Min ZHANG
;
Yi-xian LIU
;
Xin CUI
;
Yu-yan HU
;
Wen-bin LI
- Publication Type:Journal Article
- MeSH: Animals; CA1 Region, Hippocampal; metabolism; Excitatory Amino Acid Transporter 2; antagonists & inhibitors; metabolism; Ischemic Preconditioning; Oligonucleotides, Antisense; genetics; RNA, Messenger; Rats; Up-Regulation
- From: Chinese Journal of Applied Physiology 2015;31(3):238-243
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEThe present study was undertaken to design antisense oligodeoxynucleotides (AS-ODNs) of glial glutamate transporter-la (GLT-1a) and to evaluate the effectiveness of the designed AS-ODNs on the expression of GLT-1a.
METHODSFive sequences of GLT-1a AS-ODNs were designed according to the C terminus specific sequences of GLT-1a mRNA using antisense design software of IDT Com- pany. Western blot analysis was used to evaluate the inhibition effects of the five GLT-1a AS-ODNs on the expression of GLT-la.
RESULTSThe sequence of GLT-1a AS-ODNs with sequence of 5'-GGTTCTTCCTCAACACTGCA-3' could specifically inhibit the expression of GLT-1a in the hippocampal CA1 subfield of rats, while it had no effect on the expression of GLT-1b. This sequence showed similar inhibition on the expression of GLT-la in sham and ceftriaxone (Cef)-treated rats. It could also significantly inhibit the cerebral ischemic preconditioning (CIP)-induced up-regulation in the expression of GLT-1a. The magnitude of the inhibition in sham, Cef- or CIP-treated rats was similar by more than 60%.
CONCLUSIONFrom the designed five sequences of GLT-1a AS-ODNs, we obtained an effective sequence which can specifically inhibit the expression of GLT-1a.