PMID- 18276015
OWN - NLM
STAT- MEDLINE
DCOM- 20080718
LR  - 20080331
IS  - 0165-0270 (Print)
IS  - 0165-0270 (Linking)
VI  - 170
IP  - 1
DP  - 2008 May 15
TI  - Development of an in vivo adeno-associated virus-mediated siRNA approach to 
      knockdown tyrosine hydroxylase in the lateral retrochiasmatic area of the ovine 
      brain.
PG  - 56-66
LID - 10.1016/j.jneumeth.2007.12.018 [doi]
AB  - We developed a new technique of gene knockdown (KD) in a specific brain area of 
      the ewe using an adeno-associated virus (AAV)-mediated short interfering RNA 
      (siRNA) method to elucidate the importance of key factors of seasonal 
      reproduction. Two 19-nucleotide sequences (TH1 or TH2) were chosen from the 
      tyrosine hydroxylase (TH) gene. TH1, TH2 or a random sequence (TH3) was 
      incorporated into an eGFP expressing AAV vector. Firstly, 5 microl of AAV-TH1 or 
      AAV-TH2 solutions (8-9 x 10(11)Vg/ml) were stereotaxically injected into one A15 
      nucleus while the other received a control treatment. Ewes were killed after 15 
      or 75 days. The number of TH neurons was 49% and 36% lower on the AAV-TH1 treated 
      side than on the control side 15 and 75 days post-injection, respectively. 
      AAV-TH2 did not induce a significant variation in TH cell population. Finally, in 
      order to increase the KD, two groups of ewes received 10 microl of AAV-TH1 either 
      in a bolus injection or in two 5 microl inoculations carried out 2 weeks apart. 
      Only ewes receiving a bolus injection showed a larger KD reaching 66% 2 months 
      after inoculation. This method proved effective in reducing TH expression and 
      will be further developed to understand cellular mechanisms driving seasonal 
      functions.
FAU - Dufourny, Laurence
AU  - Dufourny L
AD  - UMR85 Physiologie de la Reproduction et des Comportements, Institut National de 
      la Recherche Agronomique, F-37380 Nouzilly, France. dufourny@tours.inra.fr
FAU - Migaud, Martine
AU  - Migaud M
FAU - Thiery, Jean-Claude
AU  - Thiery JC
FAU - Malpaux, Benoit
AU  - Malpaux B
LA  - eng
PT  - Journal Article
DEP - 20080106
PL  - Netherlands
TA  - J Neurosci Methods
JT  - Journal of neuroscience methods
JID - 7905558
RN  - 0 (RNA, Small Interfering)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - EC 1.14.16.2 (Tyrosine 3-Monooxygenase)
SB  - IM
MH  - Adenoviridae/*genetics
MH  - Animals
MH  - Animals, Genetically Modified
MH  - Brain/anatomy & histology/*enzymology
MH  - Female
MH  - *Gene Transfer Techniques
MH  - Genetic Vectors
MH  - Green Fluorescent Proteins/genetics
MH  - Optic Chiasm/anatomy & histology/*enzymology
MH  - Ovariectomy
MH  - RNA, Small Interfering/*pharmacology
MH  - Reproduction/physiology
MH  - Seasons
MH  - Sheep
MH  - Tyrosine 3-Monooxygenase/*genetics
EDAT- 2008/02/16 09:00
MHDA- 2008/07/19 09:00
CRDT- 2008/02/16 09:00
PHST- 2007/08/23 00:00 [received]
PHST- 2007/11/21 00:00 [revised]
PHST- 2007/12/22 00:00 [accepted]
PHST- 2008/02/16 09:00 [pubmed]
PHST- 2008/07/19 09:00 [medline]
PHST- 2008/02/16 09:00 [entrez]
AID - S0165-0270(08)00006-X [pii]
AID - 10.1016/j.jneumeth.2007.12.018 [doi]
PST - ppublish
SO  - J Neurosci Methods. 2008 May 15;170(1):56-66. doi: 
      10.1016/j.jneumeth.2007.12.018. Epub 2008 Jan 6.