PMID- 10837866
OWN - NLM
STAT- MEDLINE
DCOM- 20000815
LR  - 20190831
IS  - 0165-0270 (Print)
IS  - 0165-0270 (Linking)
VI  - 98
IP  - 1
DP  - 2000 May 15
TI  - The use of TaqMan RT-PCR assays for semiquantitative analysis of gene expression 
      in CNS tissues and disease models.
PG  - 9-20
AB  - TaqMan reverse transcription polymerase chain reaction (RT-PCR) is a recently 
      developed technique which allows the measurement of an accumulating PCR product 
      in real time. In the present study we have validated the use of TaqMan RT-PCR for 
      mRNA localisation studies in human and rat tissues, and for the investigation of 
      gene expression changes in CNS animal models. In human brain, D(2) receptor mRNA 
      was enriched in caudate nucleus and putamen, whilst in rat brain, highest levels 
      of D(2) receptor mRNA expression were observed in striatum and nucleus accumbens, 
      consistent with the known distribution of this receptor in basal ganglia. In a 
      rat model of permanent middle cerebral artery occlusion (pMCAO), endogenous 
      interleukin-1 receptor antagonist (IL-1ra) mRNA was upregulated over 30-fold at 
      24 h post-lesion in both striatum and cortex ipsilateral to artery occlusion. 
      Brain-derived neurotrophic factor (BDNF) mRNA was transiently upregulated 
      3.7-fold at 3 h, but not at 24 h or 3 days after induction of cortical spreading 
      depression (CSD) in rats. Our observations in these two animal models using 
      TaqMan RT-PCR were consistent with previous reports using other techniques. In 
      conclusion, TaqMan RT-PCR assays provide a rapid and reliable method for 
      semi-quantitative analysis of gene expression in the nervous system.
FAU - Medhurst, A D
AU  - Medhurst AD
AD  - Department of Neuroscience Research, SmithKline Beecham Pharmaceuticals, Third 
      Avenue, Essex, CM19 5AW, Harlow, UK. andy_medhurst-1@sbphrd.com
FAU - Harrison, D C
AU  - Harrison DC
FAU - Read, S J
AU  - Read SJ
FAU - Campbell, C A
AU  - Campbell CA
FAU - Robbins, M J
AU  - Robbins MJ
FAU - Pangalos, M N
AU  - Pangalos MN
LA  - eng
PT  - Journal Article
PL  - Netherlands
TA  - J Neurosci Methods
JT  - Journal of neuroscience methods
JID - 7905558
RN  - 0 (Actins)
RN  - 0 (DNA Primers)
RN  - 0 (RNA, Messenger)
RN  - 0 (Receptors, Dopamine D2)
RN  - EC 1.2.1.- (Glyceraldehyde-3-Phosphate Dehydrogenases)
RN  - EC 2.7.7.- (Taq Polymerase)
RN  - EC 5.2.1.8 (Peptidylprolyl Isomerase)
SB  - IM
MH  - Actins/genetics
MH  - Animals
MH  - Brain Chemistry/*genetics
MH  - Cortical Spreading Depression
MH  - DNA Primers
MH  - Disease Models, Animal
MH  - Gene Expression/*physiology
MH  - Glyceraldehyde-3-Phosphate Dehydrogenases/genetics
MH  - Humans
MH  - Infarction, Middle Cerebral Artery/*genetics/physiopathology
MH  - Peptidylprolyl Isomerase/genetics
MH  - RNA, Messenger/analysis
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Receptors, Dopamine D2/genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Sensitivity and Specificity
MH  - Taq Polymerase
EDAT- 2000/06/06 09:00
MHDA- 2000/08/19 11:00
CRDT- 2000/06/06 09:00
PHST- 2000/06/06 09:00 [pubmed]
PHST- 2000/08/19 11:00 [medline]
PHST- 2000/06/06 09:00 [entrez]
AID - S0165-0270(00)00178-3 [pii]
AID - 10.1016/s0165-0270(00)00178-3 [doi]
PST - ppublish
SO  - J Neurosci Methods. 2000 May 15;98(1):9-20. doi: 10.1016/s0165-0270(00)00178-3.