Cytochrome P450 aromatase in grey mullet: cDNA and promoter isolation; brain, pituitary and ovarian expression during puberty

Nocillado, Josephine N., Elizur, Abigail, Avitan, Ayelet, Carrick, Frank and Levavi-Sivan, Berta (2007) Cytochrome P450 aromatase in grey mullet: cDNA and promoter isolation; brain, pituitary and ovarian expression during puberty. Molecular and Cellular Endocrinology, 263 1-2: 65-78. doi:10.1016/j.mce.2006.08.013


Author Nocillado, Josephine N.
Elizur, Abigail
Avitan, Ayelet
Carrick, Frank
Levavi-Sivan, Berta
Title Cytochrome P450 aromatase in grey mullet: cDNA and promoter isolation; brain, pituitary and ovarian expression during puberty
Journal name Molecular and Cellular Endocrinology   Check publisher's open access policy
ISSN 0303-7207
Publication date 2007-01
Year available 2006
Sub-type Article (original research)
DOI 10.1016/j.mce.2006.08.013
Volume 263
Issue 1-2
Start page 65
End page 78
Total pages 14
Editor I. T. Huhtaniemi
W. E. Rainey
Place of publication Clare, ireland
Publisher Elsevier Ireland Ltd
Collection year 2008
Language eng
Subject C1
279999 Biological Sciences not elsewhere classified
780105 Biological sciences
Abstract In a study towards elucidating the role of aromatases during puberty in female grey mullet, the cDNAs of the brain (muCyp19b) and ovarian (muCyp19a) aromatase were isolated by RT-PCR and their relative expression levels were determined by quantitative real-time RT-PCR. The muCyp19a ORF of 1515 bp encoded 505 predicted amino acid residues, while that of muCyp19b was 1485 bp and encoded 495 predicted amino acid residues. The expression level of muCyp19b significantly increased in the brain as puberty advanced; however, its expression level in the pituitary increased only slightly with pubertal development. In the ovary, the muCyp19a expression level markedly increased as puberty progressed. The promoter regions of the two genes were also isolated and their functionality evaluated in vitro using luciferase as the reporter gene. The muCyp19a promoter sequence (650bp) contained a consensus TATA box and putative transcription factor binding sites, including two half EREs, an SF-1, an AhR/Arnt, a PR and two GATA-3s. The muCyp19b promoter sequence (2500 bp) showed consensus TATA and CCAAT boxes and putative transcription binding sites, namely: a PR, an ERE, a half ERE, a SP-1, two GATA-binding factor, one half GATA-1, two C/EBPs, a GRE, a NFkappaB, three STATs, a PPAR/RXR, an Ahr/Arnt and a CRE. Basal activity of serially deleted promoter constructs transiently transfected into COS-7, alpha T3 and TE671 cells demonstrated the enhancing and silencing roles of the putative transcription factor binding sites. Quinpirole, a dopamine agonist, significantly reduced the promoter activity of muCyp19b in TE671. The results suggest tissue-specific regulation of the muCyp19 genes and a putative alternative promoter for muCyp19b. (c) 2006 Elsevier Ireland Ltd. All rights reserved.
Keyword Cell Biology
Endocrinology & Metabolism
promoter
dopamine
estrogen
real-time quantitative RT-PCR
mullet
Bass Dicentrarchus-labrax
Halibut Hippoglossus-hippoglossus
Tilapia Oreochromis-niloticus
Immunosorbent-assay Elisa
Ribonucleic-acid Levels
Medaka Oryzias-latipes
Beta-casein Gene
Dopamine-receptor
Differential Expression
Clarias-gariepinus
Q-Index Code C1
Q-Index Status Confirmed Code

 
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Created: Mon, 18 Feb 2008, 16:02:50 EST