PMID- 14659827
OWN - NLM
STAT- MEDLINE
DCOM- 20040301
LR  - 20190901
IS  - 0165-0270 (Print)
IS  - 0165-0270 (Linking)
VI  - 131
IP  - 1-2
DP  - 2003 Dec 30
TI  - Quantification of layer-specific gene expression in the hippocampus: effective 
      use of laser microdissection in combination with quantitative RT-PCR.
PG  - 83-91
AB  - Laser microdissection in combination with quantitative RT-PCR is now widely 
      appreciated as an excellent tool for quantifying mRNA levels in defined cell 
      populations. It may be particularly useful in the hippocampal formation, where 
      principal cells form distinct and readily identifiable cell layers. Here we are 
      presenting an optimized protocol for labeling hippocampal principal cells on 
      foil-mounted sections for microdissection with the Leica AS LMD system and 
      discuss potential further applications and pitfalls. Employing this optimized 
      method, we studied changes in brain-derived neurotrophic factor (BDNF) mRNA 
      expression in granule cells of the mouse dentate gyrus following unilateral 
      entorhinal cortex lesion. In this lesioning paradigm, changes in BDNF mRNA 
      expression have previously been reported in the rat. Using laser microdissection, 
      the granule cell layers ipsi- and contralateral to the lesion were collected and 
      changes in BDNF levels were quantified using quantitative RT-PCR. BDNF mRNA 
      levels were five-fold higher on the ipsilateral side compared to levels found on 
      the contralateral side or in controls. The development of this optimized method 
      for laser microdissection and subsequent quantitative RT-PCR allows 
      layer-specific quantification of gene expression levels in the hippocampus and 
      may be similarly employed in other brain areas or tissues with a laminar 
      arrangement or high density of cells.
FAU - Burbach, Guido J
AU  - Burbach GJ
AD  - Institute of Clinical Neuroanatomy, Johann Wolfgang Goethe-University, 
      Theodor-Stern-Kai 7, D-60590 Frankfurt am Main, Germany. 
      Burbach@em.uni-frankfurt.de
FAU - Dehn, Doris
AU  - Dehn D
FAU - Del Turco, Domenico
AU  - Del Turco D
FAU - Deller, Thomas
AU  - Deller T
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - J Neurosci Methods
JT  - Journal of neuroscience methods
JID - 7905558
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (RNA, Messenger)
RN  - 0 (RNA, Ribosomal, 18S)
SB  - IM
MH  - Animals
MH  - Brain-Derived Neurotrophic Factor/genetics/metabolism
MH  - Gene Expression
MH  - Hippocampus/cytology/*metabolism
MH  - Histocytochemistry/methods
MH  - *Lasers
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Microdissection/*methods
MH  - RNA, Messenger/*analysis/biosynthesis
MH  - RNA, Ribosomal, 18S/genetics/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction/*methods
EDAT- 2003/12/09 05:00
MHDA- 2004/03/03 05:00
CRDT- 2003/12/09 05:00
PHST- 2003/12/09 05:00 [pubmed]
PHST- 2004/03/03 05:00 [medline]
PHST- 2003/12/09 05:00 [entrez]
AID - S0165027003002322 [pii]
AID - 10.1016/s0165-0270(03)00232-2 [doi]
PST - ppublish
SO  - J Neurosci Methods. 2003 Dec 30;131(1-2):83-91. doi: 
      10.1016/s0165-0270(03)00232-2.