Identification of neuroendocrine-specific protein as an ethanol-regulated gene with mRNA differential display.
Chronic ethanol exposure produces changes in behavior that may result from effects of ethanol on gene expression. To identify potentially ethanol-regulated genes, mRNA differential display was used to screen the expressed genes in whole brain of mice chronically exposed to ethanol vapors. Mice genetically selected for susceptibility (Withdrawal Seizure-Prone; WSP) or resistance (Withdrawal Seizure-Resistant; WSR) to ethanol withdrawal convulsions were exposed to either ethanol vapor (ETOH group) or air (CTL group) for 72 h. A putative ethanol-regulated product was isolated; nucleotide sequence analysis of this product revealed >85% nucleotide identity to human neuroendocrine-specific protein (NSP) gene. Northern analysis of the expression of this product revealed hybridization to two transcripts ( approximately 3.0 kb and 1.4 kb) on blots containing whole brain RNA, consistent with the transcript sizes of hNSP. Ethanol-induced regulation of mNSP was suggested in whole brain of WSP mice, but not in WSR mice, by Northern blot analysis. One transcript (3.0 kb) suggests a 26% increase in relative abundance in whole brain of ethanol-exposed WSP mice, while there was no effect of ethanol on abundance of the 1.4-kb transcript in WSP mice. No effects of ethanol were observed for WSR mice. These preliminary findings suggest that mNSP represents a novel ethanol-induced gene in mice selected for genetic susceptibility to severe ethanol withdrawal.
Schafer
G L
GL
Oregon Health Sciences University, Portland Alcohol Research Center, and Veterans Affairs Medical Center, Portland, Oregon 97201, USA.
Crabbe
J C
JC
Wiren
K M
KM
D000818:Animals; D015152:Blotting, Northern; D002492:Central Nervous System Depressants / Q000494:pharmacology*; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D000431:Ethanol / Q000494:pharmacology*; D005786:Gene Expression Regulation; D005796:Genes / Q000187:drug effects* / Q000235:genetics; D008297:Male; D051379:Mice; D009419:Nerve Tissue Proteins / Q000187:drug effects* / Q000235:genetics; D012333:RNA, Messenger / Q000187:drug effects* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment
D002492:Central Nervous System Depressants; D018076:DNA, Complementary; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; C489914:Rtn1 protein, mouse; D000431:Ethanol