Use of laser-capture microdissection for the identification of marker genes for the ventromedial hypothalamic nucleus Journal Article
| Authors: | Segal, J. P.; Stallings, N. R.; Lee, C. E.; Zhao, L.; Socci, N.; Viale, A.; Harris, T. M.; Soares, M. B.; Childs, G.; Elmquist, J. K.; Parker, K. L.; Friedman, J. M. |
| Article Title: | Use of laser-capture microdissection for the identification of marker genes for the ventromedial hypothalamic nucleus |
| Abstract: | The ventromedial hypothalamic nucleus (VMH) plays an important role in the control of feeding and energy homeostasis. In contrast to other hypothalamic nuclei that are also known to regulate energy balance, there is a paucity of nucleus-specific marker genes for the VMH, limiting the application of molecular approaches for analyzing VMH information processing, function, and circuitry. Here, we report the use of laser-capture microdissection to isolate a set of cDNAs that are enriched in the VMH relative to two adjacent hypothalamic nuclei, the arcuate and dorsomedial hypothalamus. The relative expression levels of nine of the 12 most robustly expressed VMH-enriched genes were confirmed by real-time PCR analysis using separate RNAs from these three nuclei. Three of these VMH-enriched genes were further characterized by in situ hybridization histochemistry, including pituitary adenylate cyclase activating polypeptide, cerebellin 1, and an expressed sequence tag named LBH2. Finally, to test whether some of these genes were coordinately regulated, we monitored their expression in steroidogenic factor 1 (SF-1) knock-out mice. SF-1 is a transcription factor that controls the development of the VMH. The RNA levels for four of these genes were reduced in these knock-out animals, further suggesting that they are direct or indirect targets of this orphan nuclear receptor. The VMH-enriched genes identified here provide a basis for a functional analysis of VMH neuronal subpopulations via the use of bacterial artificial chromosome transgenics and related technologies. These results also demonstrate the utility of laser-capture microdissection coupled with microarray technology to identify nucleus-specific transcriptional networks. Copyright © 2005 Society for Neuroscience. |
| Keywords: | controlled study; unclassified drug; nonhuman; flow cytometry; biological markers; mouse; animals; mice; mice, knockout; animal tissue; gene expression; nerve tissue proteins; protein; transcription factor; homeodomain proteins; mice, inbred c57bl; transcription factors; rna; gene expression regulation; in situ hybridization; reverse transcriptase polymerase chain reaction; rna, messenger; microarray analysis; expressed sequence tag; gene identification; nucleotide sequence; lasers; hypothalamus; histochemistry; real time polymerase chain reaction; microarray; blotting, northern; marker gene; knockout mouse; complementary dna; microdissection; receptors, cytoplasmic and nuclear; protein precursors; expression; marker; laser capture microdissection; lcm; ventromedial; vmh; cerebellin 1; hypophysis adenylate cyclase activating polypeptide; lbh2 protein; steroidogenic factor 1; hypothalamus nucleus; hypothalamus ventromedial nucleus; ventromedial hypothalamic nucleus |
| Journal Title: | The Journal of Neuroscience |
| Volume: | 25 |
| Issue: | 16 |
| ISSN: | 0270-6474 |
| Publisher: | Society for Neuroscience |
| Date Published: | 2005-04-20 |
| Start Page: | 4181 |
| End Page: | 4188 |
| Language: | English |
| DOI: | 10.1523/jneurosci.0158-05.2005 |
| PUBMED: | 15843621 |
| PROVIDER: | scopus |
| PMCID: | PMC6724958 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 44" - "Export Date: 24 October 2012" - "CODEN: JNRSD" - "Molecular Sequence Numbers: GENBANK: AA030506, AA982708, AB010149, AB025411, AF144629, AF174583, AF336850, BC024787, BC055730, S65878, X76653, XM_354697;" - "Source: Scopus" |
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