title: Comparison of global brain gene expression profiles between inbred long-sleep and inbred short-sleep mice by high-density gene array hybridization.
abstract: BACKGROUND
  Inbred long-sleep (ILS) and short-sleep (ISS) mice show significant central nervous system-mediated differences in sleep time for sedative dose of ethanol and are frequently used as a rodent model for ethanol sensitivity. In this study, we have used complementary DNA (cDNA) array hybridization methodology to identify genes that are differentially expressed between the brains of ILS and ISS mice.
  
  
  METHODS
  To carry out this analysis, we used both the gene discovery array (GDA) and the Mouse GEM 1 Microarray. GDA consists of 18,378 nonredundant mouse cDNA clones on a single nylon filter. Complex probes were prepared from total brain mRNA of ILS or ISS mice by using reverse transcription and 33P labeling. The labeled probes were hybridized in parallel to the gene array filters. Data from GDA experiments were analyzed with SQL-Plus and Oracle 8. The GEM microarray includes 8,730 sequence-verified clones on a glass chip. Two fluorescently labeled probes were used to hybridize a microarray simultaneously. Data from GEM experiments were analyzed by using the GEMTools software package (Incyte). Differentially expressed genes identified from each method were confirmed by relative quantitative reverse transcription-polymerase chain reaction (RT-PCR).
  
  
  RESULTS
  A total of 41 genes or expressed sequence tags (ESTs) display significant expression level differences between brains of ILS and ISS mice after GDA, GEM1 hybridization, and quantitative RT-PCR confirmation. Among them, 18 clones were expressed higher in ILS mice, and 23 clones were expressed higher in ISS mice. The individual gene or EST's function and mapping information have been analyzed.
  
  
  CONCLUSIONS
  This study identified 41 genes that are differentially expressed between brains of ILS and ISS mice. Some of them may have biological relevance in mediation of phenotypic variation between ILS and ISS mice for ethanol sensitivity. This study also demonstrates that parallel gene expression comparison with high-density cDNA arrays is a rapid and efficient way to discover potential genes and pathways involved in alcoholism and alcohol-related physiologic processes.
authors: Department of Pharmacology, University of Colorado Health Sciences Center, Denver Colorado 80262, USA .
authors: Y
authors: Y
authors: Xu
authors: M
authors: M
authors: Ehringer
authors: F
authors: F
authors: Yang
authors: J M
authors: JM
authors: Sikela
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D000954:Antigens, Surface / Q000235:genetics; D001921:Brain / Q000378:metabolism*; D018076:DNA, Complementary / Q000032:analysis; D004798:Enzymes / Q000235:genetics; D000431:Ethanol / Q000494:pharmacology; D015870:Gene Expression; D016147:Genes, Tumor Suppressor; D007473:Ion Channels / Q000235:genetics; D051379:Mice; D008815:Mice, Inbred Strains; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D009857:Oncogenes / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012890:Sleep / Q000235:genetics*; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics
chemical_list: D000954:Antigens, Surface; D018076:DNA, Complementary; D004798:Enzymes; D007473:Ion Channels; D014157:Transcription Factors; D000431:Ethanol