title: Differential expression of corticotropin-releasing factor (CRF) and urotensin I precursor genes, and evidence of CRF gene expression regulated by cortisol in goldfish brain.
abstract: Corticotropin-releasing factor (CRF) and urotensin I (UI) precursor cDNAs were cloned and sequenced from a goldfish brain cDNA library in order to investigate the distribution of CRF and UI mRNAs in goldfish brain and the regulation of CRF and UI gene expression. The CRF (966-bp) and UI (769-bp) cDNAs encode 163- and 146-amino acid precursors, respectively, and consist of a signal peptide sequence, a cryptic region, and a 41-amino acid mature peptide at the carboxy terminal. The deduced amino acid sequences of the CRF and UI peptides exhibit a sequence identity of 54%. Northern blot analysis revealed a single size of CRF (1.3 kb) and UI (2.0 kb) mRNAs, which are expressed in the telencephalon-preoptic, hypothalamic, optic tectum-thalamus, and posterior brain regions, but not in the pituitary. In addition, while the CRF gene is strongly expressed in the olfactory bulbs, the UI gene is not. In brain regions in which both genes are expressed, the mRNA levels of CRF were three- to sevenfold higher that those of UI. While the low expression levels of the UI gene prevented further analysis of its regulation, the regulation of CRF gene expression by cortisol was examined. In response to intraperitoneal implants of cortisol (300 microg/g BW) the level of CRF mRNA in the telencephalon-preoptic region decreased to 69% of control values at 6 and 24 h posttreatment. In sham-treated fish, in parallel with a transient injection stress-elicited increase in plasma cortisol, CRF mRNA levels declined to 72% of control value at 6 h postinjection and recovered after 24 h. Injection of the glucocorticoid antagonist, RU-486 (100 microg/g BW), prevented the reduction in CRF gene expression associated with the injection stress at 6 h and increased CRF mRNA levels to 145% of control value after 24 h. In contrast, the various implants had no effect on CRF mRNA levels in either the hypothalamus or the optic tectum-thalamus region. These results provide evidence of differential expression of the CRF and UI genes in hypothalamic and extrahypothalamic regions of goldfish brain. Furthermore, they demonstrate that stress levels of plasma cortisol can lead to a decrease in CRF gene expression that is mediated by glucocorticoid receptors in the telencephalon-preoptic region and give an indication of the regional specificity of the regulation of CRF gene expression by cortisol.
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Alberta, T6G 2E9, Canada.
authors.forename: N J
authors.initials: NJ
authors.lastname: Bernier
authors.forename: X
authors.initials: X
authors.lastname: Lin
authors.forename: R E
authors.initials: RE
authors.lastname: Peter
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D001923:Brain Chemistry; D003001:Cloning, Molecular; D003346:Corticotropin-Releasing Hormone / Q000235:genetics*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D006054:Goldfish / Q000235:genetics*; D006727:Hormone Antagonists / Q000494:pharmacology; D006801:Humans; D006854:Hydrocortisone / Q000494:pharmacology*; D008297:Male; D015735:Mifepristone / Q000494:pharmacology; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D017385:Sequence Homology; D014579:Urotensins / Q000235:genetics*
chemical_list: D006727:Hormone Antagonists; D011498:Protein Precursors; D012333:RNA, Messenger; D014579:Urotensins; D015735:Mifepristone; D003346:Corticotropin-Releasing Hormone; C029676:urotensin I; D006854:Hydrocortisone