GASTROINTESTINAL SIGNIFICANCE OF GASTRIN REGULATION
胃泌素调节的胃肠意义
基本信息
- 批准号:3232637
- 负责人:
- 金额:$ 23.77万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1984
- 资助国家:美国
- 起止时间:1984-07-01 至 1989-06-30
- 项目状态:已结题
- 来源:
- 关键词:Zollinger Ellison syndrome amidation /deamidation carboxyl group cell transformation clone cells complementary DNA fibroblasts gastrins genetic translation hormone biosynthesis hormone regulation /control mechanism laboratory rat messenger RNA methylprednisolone orphan disease /drug peptic ulcer secretion thymidine kinase tissue /cell culture
项目摘要
Although the physiological importance of gastrin in regulating gastric acid
secretion has been established, and thus the relationship between gastrin
and the pathogenesis of peptic ulcer disease implied, little is known about
the mechanisms by which the synthesis of this gastrointestinal hormone is
regulated. Recent development of molecular biological and cell culture
techniques has made it possible to examine the control of hormone synthesis
directly. I will use these techniques to study the biosynthesis,
post-translational processing and secretion of gastrin. To determine the
structure of the human gastrin precursor, gastrin cDNA will be synthesized
from human antral and Zollinger-Ellison tumor mRNA with reverse
transcriptase using the gastrin-specific synthetic oligodeoxynucleotide
primer, d(G-A-A-G-T-C-C-A-T-C-C-A). The cDNA will be sequenced and used as
a probe for selection of full-length gastrin cDNA clones from a cDNA
library constructed using antral and Zollinger-Ellison tumor mRNA. The
biosynthesis of gastrin will be examined with a newly developed antibody
(GL-9), which recognized gastrin precursor, and with antibody 5135, which
is specific for the carboxyl terminus of gastrin. Three systems will be
used for biosynthetic studies: 1) cell-free translation of mRNA with wheat
germ extract, 2) primary cultures of isolated canine antral gastrin cells,
and 3) gastrin secreting single cell clones formed by hybridization of
Zollinger-Ellison tumor cells with thymidine kinase deficient mouse
fibroblasts. The critical step in post-translational processing of gastrin
precursor to its active form is the amidation of the carboxyl terminal
Phe. The enzyme responsible for this amidation will be identified and
purified using a synthetic probe (Tyr-Gly-Trp-Met-Asp-Phe-Gly-Arg-Arg), an
antibody specific for this probe (GL-9), and an antibody (5135) specific
for amidated gastrin. Intermediates formed during amidation will be
identified by high pressure liquid chromatography. The pathophysiological
importance of alterations in gastrin synthesis and processing will be
examined in vivo in rats treated chronically with methyl prednisolone, or
in vitro in chronically stimulated somatic cell hybrids. In the vivo
studies, induced alterations in gastrin mRNA content, biosynthesis,
processing, and release will be examined and related to the development of
gastric ulcers.
尽管胃泌素在调节胃酸中的生理重要性
已经建立了分泌,因此胃泌素与
以及消化性溃疡疾病的发病机制,人们对此知之甚少
这种胃肠激素的合成机制是
受监管的。分子生物学和细胞培养的最新进展
技术使研究激素合成的控制成为可能。
直接去吧。我将使用这些技术来研究生物合成,
胃泌素的翻译后加工和分泌。要确定
将合成人胃泌素前体的结构,胃泌素基因
人胃窦部和Zollinger-Ellison肿瘤基因的逆转录
使用胃泌素特异性合成寡核苷酸的转录酶
D(G-A-A-G-T-C-C-A-T-C-C-A)。该基因将被测序并用于
一种从cDNA中筛选胃泌素全长cDNA克隆的方法
用胃窦和Zollinger-Ellison肿瘤基因构建文库。这个
胃泌素的生物合成将用一种新开发的抗体进行检测
(gl-9),识别胃泌素前体;与抗体5135,其
是胃泌素羧基末端所特有的。三个系统将是
用于生物合成研究:1)小麦基因的无细胞翻译
胚芽提取液,2)犬胃窦胃泌素细胞原代培养,
和3)通过杂交形成分泌胃泌素的单细胞克隆
胸苷激酶缺陷小鼠的Zollinger-Ellison肿瘤细胞
成纤维细胞。胃泌素翻译后加工的关键步骤
其活性形式的前体是羧基末端的酰胺化。
Phe.负责这一酰胺化的酶将被鉴定并
用合成探针(Tyr-Gly-Trp-Met-Asp-Phe-Gly-Arg-Arg)纯化,
该探针的特异性抗体(GL-9)和特异性抗体(5135)
用于胺化胃泌素。在酰胺化过程中形成的中间体将
经高压液相色谱鉴定。病理生理学
胃泌素合成和加工中的改变的重要性将是
在长期使用甲基强的松龙治疗的大鼠体内进行检查,或
在体外,在慢性刺激的体细胞杂交。在体内
研究,诱导胃泌素mRNA含量的变化,生物合成,
将检查处理和发布,并与开发相关
胃溃疡。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Tadataka Yamada其他文献
Tadataka Yamada的其他文献
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{{ truncateString('Tadataka Yamada', 18)}}的其他基金
GASTROINTESTINAL SIGNIFICANCE OF GASTRIN REGULATION
胃泌素调节的胃肠意义
- 批准号:
3153130 - 财政年份:1984
- 资助金额:
$ 23.77万 - 项目类别:
GASTROINTESTINAL SIGNIFICANCE OF GASTRIN REGULATION
胃泌素调节的胃肠意义
- 批准号:
3232641 - 财政年份:1984
- 资助金额:
$ 23.77万 - 项目类别:
GASTROINTESTINAL SIGNIFICANCE OF GASTRIN REGULATION
胃泌素调节的胃肠意义
- 批准号:
3232640 - 财政年份:1984
- 资助金额:
$ 23.77万 - 项目类别:
GASTROINTESTINAL SIGNIFICANCE OF GASTRIN REGULATION
胃泌素调节的胃肠意义
- 批准号:
3232638 - 财政年份:1984
- 资助金额:
$ 23.77万 - 项目类别: