PMID- 20828614
OWN - NLM
STAT- MEDLINE
DCOM- 20110705
LR  - 20211028
IS  - 1095-9327 (Electronic)
IS  - 1044-7431 (Linking)
VI  - 46
IP  - 1
DP  - 2011 Jan
TI  - D-amino acid oxidase knockdown in the mouse cerebellum reduces NR2A mRNA.
PG  - 167-75
LID - 10.1016/j.mcn.2010.08.018 [doi]
AB  - Virus mediated RNA-interference (RNAi) is a powerful approach to study genes in 
      vivo. Here we report a method using lentivirus-delivered RNAi to knockdown the 
      glial enzyme, D-amino acid oxidase (DAO), in the mouse cerebellum. After initial 
      characterisation in vitro, we achieved a 40-50% reduction of DAO mRNA in the 
      cerebellum 7 and 28 days after a single injection of lentivirus encoding a 
      DAO-specific, short-hairpin RNA. Injections also decreased DAO immunoreactivity 
      (-33%). The major substrate for DAO is D-serine, an N-methyl-D-aspartate receptor 
      (NMDAR) co-agonist. Thus, we also measured whether DAO knockdown impacted on 
      d-serine, or expression of NMDAR subunits, and found that DAO RNAi led to 
      increased cerebellar D-serine levels (+77%), and decreased NMDAR subunit NR2A 
      mRNA (-22%), but did not affect NR1 or NR2C mRNAs. These data demonstrate the 
      utility of lentiviruses to deliver RNAi to glial cells within the cerebellum, and 
      confirm the role of DAO in D-serine metabolism. They also provide a tool to 
      investigate DAO, an enzyme currently of considerable interest in the 
      pathophysiology and therapy of schizophrenia.
CI  - Copyright A(c) 2010 Elsevier Inc. All rights reserved.
FAU - Burnet, Philip W J
AU  - Burnet PW
AD  - Department of Psychiatry, University of Oxford, Warneford Hospital, Oxford OX3 
      7JX, UK. phil.burnet@psych.ox.ac.uk
FAU - Anderson, Patrick N
AU  - Anderson PN
FAU - Chen, Li
AU  - Chen L
FAU - Nikiforova, Natalia
AU  - Nikiforova N
FAU - Harrison, Paul J
AU  - Harrison PJ
FAU - Wood, Matthew J A
AU  - Wood MJ
LA  - eng
GR  - BB/D017912/1/BB_/Biotechnology and Biological Sciences Research Council/United 
      Kingdom
GR  - G0500822/MRC_/Medical Research Council/United Kingdom
GR  - G0801352/MRC_/Medical Research Council/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20100907
PL  - United States
TA  - Mol Cell Neurosci
JT  - Molecular and cellular neurosciences
JID - 9100095
RN  - 0 (NR2A NMDA receptor)
RN  - 0 (Protein Subunits)
RN  - 0 (RNA, Messenger)
RN  - 0 (Receptors, N-Methyl-D-Aspartate)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - 452VLY9402 (Serine)
RN  - EC 1.4.3.- (Dao1 protein, mouse)
RN  - EC 1.4.3.3 (D-Amino-Acid Oxidase)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Cerebellum/*enzymology
MH  - D-Amino-Acid Oxidase/*genetics/*metabolism
MH  - *Gene Knockdown Techniques
MH  - Genetic Vectors/genetics/metabolism
MH  - Green Fluorescent Proteins/genetics/metabolism
MH  - Lentivirus/genetics/metabolism
MH  - Male
MH  - Mental Disorders/physiopathology
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Protein Subunits/genetics/metabolism
MH  - RNA Interference
MH  - RNA, Messenger/*metabolism
MH  - Rats
MH  - Receptors, N-Methyl-D-Aspartate/*genetics/metabolism
MH  - Serine/metabolism
EDAT- 2010/09/11 06:00
MHDA- 2011/07/06 06:00
CRDT- 2010/09/11 06:00
PHST- 2010/03/23 00:00 [received]
PHST- 2010/08/27 00:00 [revised]
PHST- 2010/08/30 00:00 [accepted]
PHST- 2010/09/11 06:00 [entrez]
PHST- 2010/09/11 06:00 [pubmed]
PHST- 2011/07/06 06:00 [medline]
AID - S1044-7431(10)00209-5 [pii]
AID - 10.1016/j.mcn.2010.08.018 [doi]
PST - ppublish
SO  - Mol Cell Neurosci. 2011 Jan;46(1):167-75. doi: 10.1016/j.mcn.2010.08.018. Epub 
      2010 Sep 7.