Neuroscience 2003 Abstract
| Presentation Number: | 758.21 |
|---|---|
| Abstract Title: | Microarray analysis of fluorogold (FG)-labeled cortical and substantia nigra (SN) neurons harvested by laser capture microdissection (LCM). |
| Authors: |
MacKenzie, R. G.*1
; Yu, F.
; Yao, F.1
; Taube, D.
; Rao, D. D.
1Psychiatry and Behavioral Neurosci, Wayne State Univ. Sch. of Med., Detroit, MI |
| Primary Theme and Topics |
Techniques in Neuroscience - Molecular and genetic techniques |
| Session: |
758. Bioinformatics: Genomics Poster |
| Presentation Time: | Tuesday, November 11, 2003 1:00 PM-2:00 PM |
| Location: | Morial Convention Center - Hall F-I, Board # WW1 |
| Keywords: | dopamine, tyrosine hydroxylase, dopa decarboxylase, aldehyde dehydrogenase |
The complex mixture of cell types in brain impedes mRNA profiling of specific cellular phenotypes. We have developed approaches to label, harvest and analyze specific cellular phenotypes in the brain. Target cells are either genetically labeled (see Low et al., 2003 SFN Annual Meeting) or are labeled using a neural tracer as in this study. Cortical and dopamine-containing SN neurons were labeled by injection of FG into rat striatum. Five days post-injection rats were perfused and the brains frozen, sectioned and placed on slides. FG-labeled neurons within SN immuno-stained for tyrosine hydroxylase indicating FG had been taken up by the dopamine cells. Labeled SN and cortical cells were harvested from slides by LCM. Neurons from each site were picked, the RNA extracted and made into cDNA containing the T7 RNA polymerase promoter site. Two rounds of amplification were performed and a portion of the final aRNA was reverse transcribed to incorporate Cy-3 or Cy-5 label into cDNA. Differentially labeled cDNA derived from FG-labeled cortical or SN neurons were mixed and hybridized to Agilent Rat cDNA microarrays containing over 14,500 unique clones. Arrays were scanned using an Axon GenePix 4000B scanner and the data acquired by GenePix Pro. Data analysis indicated that tyrosine hydroxylase, dopa-decarboxylase and aldehyde dehydrogenase (all defining genes for dopamine SN neurons) were among the top ten genes differentially expressed in SN neurons relative to cortical neurons. These results validate the approach of in vivo labeling, harvesting and expression profiling of specific target cells within the brain.
Supported by DA16406-01
Sample Citation:
[Authors]. [Abstract Title]. Program No. XXX.XX. 2003 Neuroscience Meeting Planner. New Orleans, LA: Society for Neuroscience, 2003. Online.
Copyright © 2003-2026 Society for Neuroscience; all rights reserved. Permission to republish any abstract or part of any abstract in any form must be obtained in writing by SfN office prior to publication.
