肉仔鸡肠黏膜紧密连接典型蛋白mRNA表达量的测定

E. A. Garcia, Carlos Ignacio Soto Zárate, Francisco Rodolfo González Díaz, J. Bordes, G. Fariña, C. Tovar
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引用次数: 0

摘要

紧密连接主要由两种类型的蛋白质形成;claudins和occludin,这两种物质都是维持肠上皮完整性和屏障功能的基础。这种屏障功能允许营养物质的吸收,主要是通过胞吞作用;然而,在鸟类中,90%的物质被细胞旁机制吸收。尽管如此,在成年鸡肠道的不同部位存在的claudins尚不清楚,更不用说它们的功能作用了。本研究旨在通过RT-PCR测定鸡(Gallus Gallus domesticus)肠道不同区域(十二指肠、空肠、回肠、盲肠和直肠)中claudins 1、2、3、5、10、12、16和occludin的mRNA的存在。为了实现这一目标,使用了准备屠宰的7周龄公鸡和孵化16天的鸡胚(阳性对照)。对于所有处理过的样品,获得预期大小的扩增子;claudin 1(662 pb)、claudin 2(162 pb)、claudin 3(185 pb)、cliudin 5(224 pb)、cloudin 10(687 pb)、克劳迪12(738 pb),claudin 16(191 pb)和occludin(430 pb)。为了证实这些发现,对获得的扩增子进行测序,随后对NCBI进行基本比对,在所有分析样本中获得与原始序列100%的相关性。据我们所知,本工作代表了关于在7周龄家鸡的整个肠道中存在与紧密连接形成有关的主要蛋白质的mRNA的第一份书面报告。这些发现将有助于阐明每种报道的蛋白质在鸡细胞旁吸收过程中的特定功能。
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Determination of mRNA Expression of Typical Proteins of Tight Junctions in the Intestinal Mucosa of Broilers (Gallus gallus domesticus)
Tight junctions are mainly formed by two types of proteins; claudins and occludin, both of which are fundamental to maintain the integrity and barrier function of the intestinal epithelium. This barrier function allows for the absorption of nutrients, mainly by transcytosis; however, in birds, 90% of the substances are absorbed by paracellular mechanisms. Despite this, claudins present in the different parts of the intestinal tract of adult chickens are not known, much less their functional role. This study aimed to determine the presence of mRNA of claudins 1, 2, 3, 5, 10, 12, 16 and occludin, in the different regions of the intestine (duodenum, jejunum, ileum, cecum, and rectum) in chickens (Gallus gallus domesticus) through RT-PCR. To meet this goal, 7 weeks old roosters destined for slaughter and chicken embryos of 16 days of incubation (positive control) were used. For all the processed samples, amplicons of the expected size were obtained; claudin 1 (662 pb), claudin 2 (162 pb), claudin 3 (185 pb), claudin 5 (224 pb), claudin 10 (687 pb), claudin 12 (738 pb), claudin 16 (191 pb) and occludin (430 pb). To corroborate these findings, obtained amplicons were sequenced and, subsequently, a basic alignment was performed on the NCBI, obtaining a correlation of 100% with the original sequences in all analyzed samples. To our knowledge, the present work represents the first written report regarding the presence of mRNA of the main proteins involved in tight junction formation throughout the intestinal tract of domestic chickens of 7 weeks of age. These findings will allow elucidating the specific function of each of the reported proteins in the process of paracellular absorption in chickens.
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