当前位置: X-MOL 学术Microsc. Res. Tech. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Immunohistochemical detection of hepatocyte nuclear factor-4α in vertebrates
Microscopy Research and Technique ( IF 2.5 ) Pub Date : 2021-07-01 , DOI: 10.1002/jemt.23848
Shuying Jiang 1, 2 , Toshiya Tanaka 3 , Ren Yagami 4 , Go Hasegawa 5 , Hajime Umezu 6 , Yukio Fujiyoshi 7 , Tatsuhiko Kodama 3 , Makoto Naito 8 , Yoichi Ajioka 2
Affiliation  

Hepatocyte nuclear factor-4α (HNF4α) presents in multiple isoforms generated using alternative promoter (P1 and P2) and splicing. Neither conservation of tissue distribution of HNF4α isoforms, nor presence of alternative promoter usage is known. In this study, to detect the expression of HNF4α in some species of animals, we have applied monoclonal antibodies against P1 (K9218) and P2 (H6939) promoter-driven and P1/P2 promoter-driven H1415 HNF4α for immunohistochemistry and western blot analysis. Antibody K9218 was observed in the hepatocytes, proximal tubules of the kidney, and epithelial cells in the mucosa of the small intestine and colon of rats, chicken, and tortoise, whereas antibody H6939 signal were detected in the stomach, pancreas, bile duct, and pancreatic duct of human and rats. The signal for antibody K9218 was recognized in tissues of a wide range of mammals, bird, reptile, amphibian, and fish as well. Antibody H1415 displayed a positive reaction in hepatocytes and intestinal epithelial cells in chicken and tortoise, whereas the bile duct, mucosal epithelial cells in the stomach, or pancreas in these animals were negative. Western blotting showed the binding of the antibody with HNF4α protein from each animal. The sequence of human HNF4α was 100% identical to murine and rat HNF4α, 88.9% to chicken, 77.8% to Xenopus HNF4α, and 81.5% to medaka. However, the specific part of human and invertebrate Drosophila HNF4 shares only 14.8% sequence identity. This antibody is useful for detecting HNF4α isoforms in a wide range of vertebrates, and suggests many insights into animal evolution.

中文翻译:

脊椎动物肝细胞核因子-4α的免疫组化检测

肝细胞核因子 4α (HNF4α) 以使用替代启动子(P1 和 P2)和剪接产生的多种亚型存在。既不保存 HNF4α 同种型的组织分布,也不知道是否存在替代启动子使用。在本研究中,为了检测某些动物物种中 HNF4α 的表达,我们应用了针对 P1 (K9218) 和 P2 (H6939) 启动子驱动和 P1/P2 启动子驱动的 H1415 HNF4α 的单克隆抗体进行免疫组织化学和蛋白质印迹分析。在大鼠、鸡和陆龟的肝细胞、肾脏近端小管和小肠和结肠黏膜上皮细胞中观察到抗体 K9218,而在胃、胰腺、胆管和陆龟中检测到抗体 H6939 信号。人和大鼠的胰管。抗体 K9218 的信号也在多种哺乳动物、鸟类、爬行动物、两栖动物和鱼类的组织中得到识别。抗体 H1415 在鸡和陆龟的肝细胞和肠上皮细胞中呈阳性反应,而在这些动物的胆管、胃粘膜上皮细胞或胰腺中呈阴性反应。Western印迹显示抗体与来自每只动物的HNF4α蛋白结合。人HNF4α序列与鼠和大鼠HNF4α序列100%相同,与鸡88.9%,与鸡77.8%相同。Western印迹显示抗体与来自每只动物的HNF4α蛋白结合。人HNF4α序列与鼠和大鼠HNF4α序列100%相同,与鸡88.9%,与鸡77.8%相同。Western印迹显示抗体与来自每只动物的HNF4α蛋白结合。人HNF4α序列与鼠和大鼠HNF4α序列100%相同,与鸡88.9%,与鸡77.8%相同。非洲爪蟾HNF4α 和 81.5% 到青鳉。然而,人类和无脊椎动物果蝇HNF4的特定部分仅具有 14.8% 的序列同一性。该抗体可用于检测多种脊椎动物中的 HNF4α 同种型,并为动物进化提供了许多见解。
更新日期:2021-07-01
down
wechat
bug