INSULIN INDUCTION OF PROLACTIN MRNA IN GH3 CELLS
GH3 细胞中催乳素 mRNA 的胰岛素诱导
基本信息
- 批准号:3235082
- 负责人:
- 金额:$ 14.49万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1986
- 资助国家:美国
- 起止时间:1986-02-01 至 1991-01-31
- 项目状态:已结题
- 来源:
- 关键词:DNA methylation chromatin cyclic AMP gel electrophoresis genetic manipulation genetic transcription glycosyltransferase insulin insulin receptor insulinlike factor messenger RNA molecular cloning nucleoproteins peptide hormone biosynthesis phosphorylation pituitary gland plasmids prolactin tissue /cell culture triiodothyronine
项目摘要
The GH cells are rat pituitary derived cell lines (GH1, GC, GH3, and GH4)
which produce growth hormone and or prolactin in response to many
hormones. Preliminary data demonstrate that insulin and triiodothyronine
(T3) under serum free conditions induce prolactin synthesis and prolactin
mRNA levels 3- to 14-fold in GH3 cells. Together these hormones induce a
dramatic 20- to 100-fold synergistic response. First, this research will
attempt to define this response. The kinetics will be determined by in
vitro transcription and in vivo [3H]uridine labeling studies using filter
hybridization to determine the levels of total prolactin specific
sequences. The specificity of the response for insulin or other
insulin-like growth factors (IGF) will then be determined by dose response
studies with insulin, insulin analogs, IGF-I and IGF-II. Studies to
determine the effects of insulin and T3 on prolactin RNA processing and
half-life are also proposed. Finally, the deinduction of the response
after hormone removal will be investigated. Second, we will attempt to
define possible insulin regulation of prolactin mRNA accumulation by
studying effects of insulin and T3 on cis-active DNA regulatory sequences,
trans-active regulatory molecules, or chromatin structure. Cis-active
regulatory sequences will be studied by construction of a chimeric plasmid
of 5' prolactin genomic DNA and the bacterial xanthine-guanine
phosphoribosyl transferase (XGPRT) gene and studying hormonal regulation of
XGPRT in cells transfected with this plasmid and with deletion mutants of
this plasmid. Attempts will be made to identify any role of trans-acting
regulatory molecules on this system by inhibitor studies and by two
dimensional gel electrophoresis of labeled nuclear proteins. Finally, we
will attempt to determine hormonal effects on the chromatin structure of
the prolactin gene by DNase I sensitivity studies and examination of the
methylatin state of the gene.
GH细胞是大鼠垂体衍生的细胞系(GH 1、GC、GH 3和GH4)
它们产生生长激素和/或催乳素,
荷尔蒙 初步数据表明胰岛素和三碘甲状腺原氨酸
(T3)在无血清条件下诱导催乳素合成,
在GH 3细胞中mRNA水平为3- 14倍。 这些激素共同诱导
惊人的20到100倍的协同反应。 首先,这项研究将
尝试定义这个答案。 动力学将通过以下方式确定:
使用滤膜进行体外转录和体内[3 H]尿苷标记研究
杂交以确定总催乳素特异性
序列的 对胰岛素或其他药物反应的特异性
然后通过剂量反应测定胰岛素样生长因子(IGF)
胰岛素、胰岛素类似物、IGF-I和IGF-II的研究。 研究来
确定胰岛素和T3对催乳素RNA加工的影响,
还提出了半衰期。 最后,反应的反诱导
激素去除后将进行调查。 第二,我们将努力
定义胰岛素对催乳素mRNA积累的可能调节,
研究胰岛素和T3对顺式活性DNA调控序列的影响,
反式调节分子或染色质结构。 顺式活性
将通过构建嵌合质粒来研究调控序列
5'催乳素基因组DNA和细菌黄嘌呤-鸟嘌呤
磷酸核糖转移酶(XGPRT)基因和研究激素调节
XGPRT在用该质粒和用XGPRT的缺失突变体转染的细胞中的表达
这个质粒 将尝试确定任何作用的trans-acting
通过抑制剂研究和两个
标记的核蛋白的三维凝胶电泳。 最后我们
将试图确定激素对染色质结构的影响
催乳素基因的DNA酶I敏感性研究和检查
基因的甲基化状态。
项目成果
期刊论文数量(4)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Stimulation of prolactin gene expression by insulin.
- DOI:10.1016/s0021-9258(18)37725-1
- 发表时间:1988-09
- 期刊:
- 影响因子:0
- 作者:F. Stanley
- 通讯作者:F. Stanley
The homeodomain protein, Pit-1/GHF-1, is capable of binding to and activating cell-specific elements of both the growth hormone and prolactin gene promoters.
同源域蛋白 Pit-1/GHF-1 能够结合并激活生长激素和催乳素基因启动子的细胞特异性元件。
- DOI:10.1210/mend-4-7-1069
- 发表时间:1990
- 期刊:
- 影响因子:0
- 作者:Fox,SR;Jong,MT;Casanova,J;Ye,ZS;Stanley,F;Samuels,HH
- 通讯作者:Samuels,HH
Transcriptional regulation of prolactin gene expression by thyroid hormone--alternate suppression and stimulation in different GH cell lines.
甲状腺激素对催乳素基因表达的转录调节——不同 GH 细胞系中的交替抑制和刺激。
- DOI:10.1210/mend-3-10-1627
- 发表时间:1989
- 期刊:
- 影响因子:0
- 作者:Stanley,F
- 通讯作者:Stanley,F
c-erbA protooncogenes mediate thyroid hormone-dependent and independent regulation of the rat growth hormone and prolactin genes.
c-erbA 原癌基因介导大鼠生长激素和催乳素基因的甲状腺激素依赖性和独立调节。
- DOI:10.1210/mend-2-10-902
- 发表时间:1988
- 期刊:
- 影响因子:0
- 作者:Forman,BM;Yang,CR;Stanley,F;Casanova,J;Samuels,HH
- 通讯作者:Samuels,HH
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FREDERICK M STANLEY其他文献
FREDERICK M STANLEY的其他文献
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{{ truncateString('FREDERICK M STANLEY', 18)}}的其他基金
Fox proteins mediate insulin-increasedPAI-1 transcription
Fox 蛋白介导胰岛素增加的 PAI-1 转录
- 批准号:
7140661 - 财政年份:2005
- 资助金额:
$ 14.49万 - 项目类别:
Fox proteins mediate insulin-increasedPAI-1 transcription
Fox 蛋白介导胰岛素增加的 PAI-1 转录
- 批准号:
7030592 - 财政年份:2005
- 资助金额:
$ 14.49万 - 项目类别:
INSULIN STIMULATION OF PROLACTIN GENE EXPRESSION
胰岛素刺激催乳素基因表达
- 批准号:
2142953 - 财政年份:1992
- 资助金额:
$ 14.49万 - 项目类别:
INSULIN STIMULATION OF PROLACTIN GENE EXPRESSION
胰岛素刺激催乳素基因表达
- 批准号:
3244748 - 财政年份:1992
- 资助金额:
$ 14.49万 - 项目类别:
INSULIN STIMULATION OF PROLACTIN GENE EXPRESSION
胰岛素刺激催乳素基因表达
- 批准号:
2331429 - 财政年份:1992
- 资助金额:
$ 14.49万 - 项目类别:
INSULIN STIMULATION OF PROLACTIN GENE EXPRESSION
胰岛素刺激催乳素基因表达
- 批准号:
2654506 - 财政年份:1992
- 资助金额:
$ 14.49万 - 项目类别:
INSULIN STIMULATION OF PROLACTIN GENE EXPRESSION
胰岛素刺激催乳素基因表达
- 批准号:
2142955 - 财政年份:1992
- 资助金额:
$ 14.49万 - 项目类别:
INSULIN STIMULATION OF PROLACTIN GENE EXPRESSION
胰岛素刺激催乳素基因表达
- 批准号:
3244747 - 财政年份:1992
- 资助金额:
$ 14.49万 - 项目类别:
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