A histochemical study on the expression and localization of growth factors

生长因子表达和定位的组织化学研究

基本信息

批准号：
06670014
负责人：
ISEKI Shoichi
金额：
$ 1.34万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

We have used both immunohistochemistry and in situ hybridization techniques to determine the sites of localization of immunoreactivity and mRNA signal of various growth factors in the entire rat body, and have obtained the following results :The expression of hepatocyte growth factor (HGF) was demonstrated in the submandibular gland of rats. Both the mRNA signal and immunoreactivity for HGF were localized to the epithelial cells of granular convoluted tubules. This finding added HGF to the list of many growth factors produced in the rodent submandibular gland.As an attempt for the electron microscopic in situ hybridization, the cryostat sections of rat submandibular gland were first treated for in situ hybridization with ^<35>S-labelled EGF probe and then embedded in epoxy resin for electron microscopic radioautography. The result suggested that this method is more useful for identifying morphologically the type of mRNA-positive cells than determining the subcellular localization of mRNA.An immunohistochemical study using an antibody against acidic fibroblast growth factor (aFGF) revealed that subpopulations of endocrine cells in the pancreas and small intestinal mucosa are positively immunostained in their secretory granules. This result suggested that an aFGF-like substance is produced by the cell populations that express glucagon-precursor.The expression of vascular endothelial growth factor (VEGF) is very low in the normal rat submandibular gland. However, a stimulation of submandibular gland to grow by isoproterenol caused a transient rise in the rate of expression of VEGF in this organ that peaked at 1 h after the stimulation. The mRNA was localized to the acinar cells. This result suggested a certain relation between isoproterenol-induced growth of submandibular gland and VEGF effects on the blood vessels.

我们采用免疫组化和原位杂交技术，确定了大鼠全身各生长因子的免疫反应性和mRNA信号的定位位点，并得到了以下结果：大鼠的颌下腺。 HGF 的 mRNA 信号和免疫反应性均定位于颗粒曲小管的上皮细胞。这一发现将HGF添加到啮齿动物颌下腺中产生的许多生长因子的列表中。作为电子显微镜原位杂交的尝试，首先用^ 35 S-处理大鼠颌下腺的低温切片进行原位杂交。标记 EGF 探针，然后嵌入环氧树脂中进行电子显微镜放射自显影。结果表明，该方法对于从形态学上鉴定 mRNA 阳性细胞的类型比确定 mRNA 的亚细胞定位更有用。使用抗酸性成纤维细胞生长因子 (aFGF) 的抗体进行的免疫组织化学研究表明，胰腺中内分泌细胞的亚群小肠粘膜的分泌颗粒免疫染色呈阳性。这一结果表明，aFGF样物质是由表达胰高血糖素前体的细胞群产生的。正常大鼠颌下腺中血管内皮生长因子(VEGF)的表达非常低。然而，异丙肾上腺素刺激下颌下腺生长导致该器官中 VEGF 表达率短暂升高，并在刺激后 1 小时达到峰值。 mRNA 定位于腺泡细胞。这一结果表明异丙肾上腺素诱导的颌下腺生长与VEGF对血管的作用之间存在一定的关系。