Low-density cDNA array-coupled to PCR differential display identifies new estrogen-responsive genes during the postnatal differentiation of the rat hypothalamus

被引:17
作者
Choi, EJ
Ha, CM
Choi, JG
Kang, SS
Choi, WS
Park, SK
Kim, KJ
Lee, BJ [1 ]
机构
[1] Univ Ulsan, Coll Nat Sci, Dept Biol Sci, Ulsan 680749, South Korea
[2] Gyeongsang Natl Univ, Coll Med, Dept Anat, Chinju 660280, South Korea
[3] Univ Ulsan, Coll Med, Dept Pediat, Ulsan 682060, South Korea
[4] Seoul Natl Univ, Sch Biol Sci, Seoul 151742, South Korea
来源
MOLECULAR BRAIN RESEARCH | 2001年 / 97卷 / 02期
关键词
estrogen-sterilized rat; hypothalamic sexual differentiation; differential display PCR; low-density cDNA array; female puberty;
D O I
10.1016/S0169-328X(01)00302-3
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
To identify estrogen (E)-responsive genes that may play important roles in the sexual differentiation and 1 maturation of the neuroendocrine hypothalamus, we used mRNA differential display PCR to analyze hypothalamic RNA derived from estrogen-sterilized rats (ESRs). Neonatal rats were s.c.-injected with 100 mug of 17beta-estradiol-benzoate (EB) for 5 days. Approximately 300 out of more than 2000 RNAs examined displayed a differential expression pattern between hypothalami of the ESR females compared to their 60-day-old controls. EB-dependent expression of these genes was further analyzed by low-density cDNA array using cDNA probe sets reverse-transcribed from the same groups; 98 genes were confirmed to be differentially expressed. We selected 41 clones that showed higher density differences between the two probe sets than mean density difference in control cyclophilin cDNA blots in the cDNA array. After being cloned into pGEM-T vectors, their sequences were analyzed. Homology searches identified four genes as a protein kinase C (PKC)-binding protein, NELL2 (clone 6-1), a thyroid nuclear factor, TTF-1 (9-1), Munc18-1 (17-6), and leuserpin-2 (18-5). The other 22 genes were similar to reported genes or cDNAs such as mouse kinesin-associated protein 3 (KAP3, 8b), mouse IgE binding lectin (15-1), normalized rat brain cDNA (5-1), rat cDNA (8-1) and rat embryonic cDNA (17-1). Fifteen clones such as clone 7-3 showed no match in the GenBank Database. Further characterization of eight clones (17-1, 7-3, 8-1, 5-1, NELL2, KAP3 homolog, IgE binding lectin homolog, and TTF-1) showed that their expression in the adult female rat hypothalamus is sensitive to neonatal treatment with EB. They showed brain-specific expression and moreover, showed an increase in their mRNA level before the initiation of puberty. Some of them showed gender differences in their different postnatal expression pattern. We speculate that further study will demonstrate that many of the E-regulated genes identified in the present study play important roles in the regulation of the sexual differentiation and E-dependent maturation of the hypothalamus. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:115 / 128
页数:14
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