title: Nuclei and subnuclei gene expression profiling in mammalian brain.
abstract: Information on the neuroanatomical expression of a given gene is critical to understanding its function in the central nervous system. The integration of laser capture microdissection (LCM), T7-based RNA amplification and cDNA microarrays allows for this information to be simultaneously generated for thousands of genes. To validate this integrative approach, we catalogued the gene expression profiles of seven rat brain nuclei or subnuclei. A hundred cells from the following seven brain nuclei were analyzed: locus coeruleus (LC), dorsal raphe nucleus (DR), parvocellular division (PA) and magnocellular division (MG) of the hypothalamic paraventricular nucleus (PVN) and CA1, CA3 and dentate gyrus (DG) divisions of the hippocampal formation. Of the 2145 genes investigated, 1402 genes (65%) gave a hybridization signal statistically different from the background level that was defined by non-specific hybridizations to 15 different plant genes. Validation of our microarray data on four arbitrarily selected genes was confirmed by Real-Time PCR. Previous research showing expression patterns of 'signature' genes (n=17) for specific brain nuclei are consistent with our findings. For example, as previously shown, enriched mRNA expression encoding the serotonin transporter or tyrosine hydroxylase was found in DR and LC cells, respectively. Interestingly, expression of the serotonin 5-HT(2B) receptor mRNA was also found in DR cells. We confirmed this new finding by in-situ hybridization. The hierarchical clustering analysis of gene expression shows that the two divisions of the PVN (PA and MG) are closely related to each other, as well as the three regions of the hippocampal formation (CA1, CA3 and DG), which also showed similar gene expression profiles. This study demonstrates the importance, feasibility and utility of cellular brain nuclei profiling.
authors.affiliation: Johnson & Johnson Pharmaceutical Research & Development LLC, San Diego, CA 92121, USA. pbonave1@prdus.jnj.com
authors.forename: Pascal
authors.initials: P
authors.lastname: Bonaventure
authors.forename: Hongqing
authors.initials: H
authors.lastname: Guo
authors.forename: Bin
authors.initials: B
authors.lastname: Tian
authors.forename: Xuejun
authors.initials: X
authors.lastname: Liu
authors.forename: Anton
authors.initials: A
authors.lastname: Bittner
authors.forename: Barbara
authors.initials: B
authors.lastname: Roland
authors.forename: Ranelle
authors.initials: R
authors.lastname: Salunga
authors.forename: Xiao-Jun
authors.initials: XJ
authors.lastname: Ma
authors.forename: Fredrik
authors.initials: F
authors.lastname: Kamme
authors.forename: Bernhard
authors.initials: B
authors.lastname: Meurers
authors.forename: Margot
authors.initials: M
authors.lastname: Bakker
authors.forename: Mirek
authors.initials: M
authors.lastname: Jurzak
authors.forename: Josée E
authors.initials: JE
authors.lastname: Leysen
authors.forename: Mark G
authors.initials: MG
authors.lastname: Erlander
mesh_terms: D000818:Animals; D001923:Brain Chemistry / Q000235:genetics*; D016000:Cluster Analysis; D015870:Gene Expression / Q000502:physiology; D020869:Gene Expression Profiling / Q000379:methods / Q000706:statistics & numerical data; D005819:Genetic Markers; D006624:Hippocampus / Q000737:chemistry / Q000378:metabolism*; D017403:In Situ Hybridization; D008297:Male; D008636:Mesencephalon / Q000737:chemistry / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods / Q000706:statistics & numerical data; D010286:Paraventricular Hypothalamic Nucleus / Q000737:chemistry / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar; D044403:Receptor, Serotonin, 5-HT2B; D011985:Receptors, Serotonin / Q000096:biosynthesis / Q000235:genetics
chemical_list: D005819:Genetic Markers; D012333:RNA, Messenger; D044403:Receptor, Serotonin, 5-HT2B; D011985:Receptors, Serotonin