Analysis of genes related to expression of aromatase and estradiol-regulated genes during sex differentiation in Xenopus embryos.
We analyzed the genes that exhibit transcriptional changes during sex differentiation in Xenopus, using fluorescent differential display (FDD). Search was then undertaken for sequences that were homologous to the differentially displayed DNA. In this report, trans-acting factors of activating transcription factor 4 (ATF 4) and heat shock proteins were selected, on the basis of homology, from candidate genes thought to be involved in the expression cascade of aromatase and estrogen receptor genes. The stage and tissue specificities and the effect of estradiol treatment on the expression of these genes were then examined using real-time quantitative polymerase chain reaction (RQ-RT-PCR). The expression of ATF 4, a member of the ATF/cAMP-responsive element-binding protein (CREB) family of genes, peaked in the gonads at stage 50 of development. Interestingly, expression of the genes encoding the heat shock cognate protein70. II (Hsc70. II) and the heat shock protein 70 (Hsp70) binding protein was strongly activated at stages 50 and 48 of development, respectively. The three genes revealed a higher transcription activity in the gonads than in other tissues. Although the expression of all of the genes encoding ATF 4, aromatase, Hsc70. II, and Hsp70 binding protein was activated in vitro by estrogen treatment, that of Hsc70. II and Hsp70 binding protein was found to be transient.
Akatsuka
Naoya
N
Laboratory of Biochemistry, Department of Chemistry, College of Humanities and Sciences, Nihon University, 3-25-40 Sakurajosui, Setagaya-ku, Tokyo 156-8550, Japan.
Kobayashi
Hiroshi
H
Watanabe
Emi
E
Iino
Teruhiko
T
Miyashita
Kiichi
K
Miyata
Shohei
S
D051701:Activating Transcription Factor 4; D000818:Animals; D001141:Aromatase / Q000096:biosynthesis* / Q000235:genetics; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000235:genetics / Q000502:physiology*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics / Q000502:physiology*; D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012733:Sex Differentiation / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014981:Xenopus / Q000196:embryology* / Q000235:genetics / Q000378:metabolism; D029867:Xenopus Proteins
D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; C490436:Heat shock cognate protein 70, Xenopus; D014157:Transcription Factors; D029867:Xenopus Proteins; D051701:Activating Transcription Factor 4; D004958:Estradiol; D012313:RNA; D001141:Aromatase