Neuroscience 2003 Abstract
| Presentation Number: | 758.22 |
|---|---|
| Abstract Title: | Gene expression studies in genetically-labeled hypothalamic pro-opiomelanocortin (POMC) neurons harvested by laser capture microdissection (LCM). |
| Authors: |
Granneman, J. G.*1
; Yao, F.1
; Gong, L.1
; Rao, D. D.2
; Smart, J. L.3
; Rubinstein, M.
; Low, M. J.3
; MacKenzie, R. G.1
1Dept. Psychiatry, Wayne State Univ. Sch. Med, Detroit, MI 2MI, Scott Hall, 48201, 3USA, Scott Hall, 48201, |
| Primary Theme and Topics |
Techniques in Neuroscience - Molecular and genetic techniques |
| Session: |
758. Bioinformatics: Genomics Poster |
| Presentation Time: | Tuesday, November 11, 2003 2:00 PM-3:00 PM |
| Location: | Morial Convention Center - Hall F-I, Board # WW1 |
| Keywords: | QRT-PCR, arcuate nucleus, EGFP |
In the brain, neighboring neurons are often phenotypically distinct and can subserve opposing physiological processes. For example, POMC- and NPY-expressing neurons form distinct neuronal subtypes within the arcuate nucleus and have opposite effects on hypothalamic control of appetite and body weight regulation. Interpretation of gene expression profiling of these neurons, which differentially respond to hormones and metabolic state, depends on the ability to selectively label, harvest and analyze a single neuronal subtype. We have developed an approach to label, harvest and analyze targeted neuronal subtypes in the brain. In this approach, target cells are either labeled by a neuronal tracer (see MacKenzie et al., 2003 SFN Annual Meeting) or are genetically labeled as in this study. POMC neurons were labeled by EGFP expressed by the POMC promoter in transgenic mice. Mice were perfused with 4% paraformaldehyde and cryosections prepared for LCM. EGFP-positive cells were identified by direct fluorescence and individually picked using an Arcutus PixCell II LCM system. Fifteen hundred POMC neurons could be picked from the arcuate nucleus of a single transgenic mouse. RNA extracted from the neurons was used for quantitative RT-PCR (QRT-PCR) analysis of various genes known to be expressed in these POMC neurons. Results from QRT-PCR analysis indicate that the RNA from LCM-harvested POMC neurons was intact and reproducibly amplified by QRT-PCR, thereby setting the stage for mRNA profiling of hormone or metabolic state effects on POMC neurons.
Supported by MLSC-27; DK61797; DK55819; TWO1233
<B>Conflict of Interest:</B> MJ Low and M Rubinstein have intellectual property interests in the neuronal POMC system and MJL is a recipient of discretionary gift funds from Neurocrine Biosciences, Inc.
Sample Citation:
[Authors]. [Abstract Title]. Program No. XXX.XX. 2003 Neuroscience Meeting Planner. New Orleans, LA: Society for Neuroscience, 2003. Online.
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