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.2000 Aug 15;97(17):9543-8.
doi: 10.1073/pnas.160270997.

Gene expression profiling in the human hypothalamus-pituitary-adrenal axis and full-length cDNA cloning

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Gene expression profiling in the human hypothalamus-pituitary-adrenal axis and full-length cDNA cloning

R M Hu et al. Proc Natl Acad Sci U S A..

Abstract

The primary neuroendocrine interface, hypothalamus and pituitary, together with adrenals, constitute the major axis responsible for the maintenance of homeostasis and the response to the perturbations in the environment. The gene expression profiling in the human hypothalamus-pituitary-adrenal axis was catalogued by generating a large amount of expressed sequence tags (ESTs), followed by bioinformatics analysis (http://www.chgc.sh.cn/ database). Totally, 25,973 sequences of good quality were obtained from 31,130 clones (83.4%) from cDNA libraries of the hypothalamus, pituitary, and adrenal glands. After eliminating 5,347 sequences corresponding to repetitive elements and mtDNA, 20,626 ESTs could be assembled into 9, 175 clusters (3,979, 3,074, and 4,116 clusters in hypothalamus, pituitary, and adrenal glands, respectively) when overlapping ESTs were integrated. Of these clusters, 2,777 (30.3%) corresponded to known genes, 4,165 (44.8%) to dbESTs, and 2,233 (24.3%) to novel ESTs. The gene expression profiles reflected well the functional characteristics of the three levels in the hypothalamus-pituitary-adrenal axis, because most of the 20 genes with highest expression showed statistical difference in terms of tissue distribution, including a group of tissue-specific functional markers. Meanwhile, some findings were made with regard to the physiology of the axis, and 200 full-length cDNAs of novel genes were cloned and sequenced. All of these data may contribute to the understanding of the neuroendocrine regulation of human life.

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Figures

Figure 1
Figure 1
Gene expression profile in whole HT, NP, and AD. (A) Schematic chart of ESTs from HT (8,876 ESTs), NP (7,221 ESTs), and AD (9,875 ESTs) libraries. (B) The order of gene expression levels of five classical NP hormones. Corticotropin was grouped into POMC. Real-time PCR performed for GH, POMC, and TSH generated gene expression levels (32 × 105, 5.9 × 105, and 2.4 × 105 copies/100 ng RNA, respectively,P value all < 0.001) proportional to those estimated by EST copy number. (C) The order of steroids biosynthesis-related enzymes and proteins in AD. StAR, steroidogenic acute regulatory protein; 3βHSD, type II 3β hydroxysteroid dehydrogenase/5-δ-4-δ isomerase; P450 17α, cytochrome P450, subfamily XVII (steroid 17α-hydroxylase); P45011b2, cytochrome P450 XIB2 (aldosterone synthase); ADR, adrenodoxin; P45°C21B, 21-hydroxylase B; P45011β, cytochrome P450 11β; P450scc, cholesterol side-chain cleavage enzyme P450scc; STD, dehydroepiandrosterone sulfotransferase; 17βHSD: 17-β-hydroxysteroid dehydrogenase; P45°C21, steroid 21-hydroxylase [P450(C21)]; P45021A, 21-hydroxylase A; EST, estrogen sulfotransferase. (D) Overlapping of EST clusters corresponding to known genes, known ESTs, and novel ESTs expressed in HT, NP, and AD.
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