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肉牛小肠上皮细胞中核苷转运体mRNA的基础表达在不同个体间存在差异,且瘤胃或皱胃灌注淀粉水解产物会使其发生不同程度的改变。

Basal expression of nucleoside transporter mRNA differs among small intestinal epithelia of beef steers and is differentially altered by ruminal or abomasal infusion of starch hydrolysate.

作者信息

Liao S F, Alman M J, Vanzant E S, Miles E D, Harmon D L, McLeod K R, Boling J A, Matthews J C

机构信息

Department of Animal and Food Sciences, University of Kentucky, Lexington, KY 40546, USA.

出版信息

J Dairy Sci. 2008 Apr;91(4):1570-84. doi: 10.3168/jds.2007-0763.

Abstract

In ruminants, microbial-derived nucleic acids are a major source of N and are absorbed as nucleosides by small intestinal epithelia. Although the biochemical activities of 2 nucleoside transport systems have been described for cattle, little is known regarding the regulation of their gene expression. This study was conducted to test 2 hypotheses: (1) the small intestinal epithelia of beef cattle differentially express mRNA for 3 concentrative (CNT1, 2, 3) and 2 equilibrative (ENT1, 2) nucleoside transporters (NT), and (2) expression of these NT is responsive to small intestine luminal supply of rumen-derived microbes (hence, nucleosides), energy (cornstarch hydrolysate, SH), or both. Eighteen ruminally and abomasally catheterized Angus steers (260 +/- 17 kg of BW) were fed an alfalfa cube-based diet at 1.33x NE(m) requirement. Six steers in each of 3 periods were blocked by BW (heavy vs. light). Within each block, 3 steers were randomly assigned to 3 treatments (n = 6): ruminal and abomasal water infusion (control), ruminal SH infusion/abomasal water infusion, or ruminal water infusion/abomasal SH infusion. The dosage of SH infusion amounted to 20% of ME intake. After a 14-or 16-d infusion period, steers were slaughtered, and duodenal, jejunal, and ileal epithelia were harvested for total RNA extraction and the relative amounts of mRNA expressed were determined using real-time RT-PCR quantification methodologies. All 5 NT mRNA were found expressed by each epithelium, but their abundance differed among epithelia. Specifically, jejunal expression of all 5 NT mRNA was higher than that by the ileum, whereas jejunal expression of CNT1, CNT3, and ENT1 mRNA was higher, or tended to be higher, than duodenal expression. Duodenal expression of CNT2, CNT3, and ENT2 mRNA was higher than ileal expression. With regard to SH infusion treatments, ruminal infusion increased duodenal expression of CNT3 (67%), ENT1 (51%), and ENT2 (39%) mRNA and ileal expression of CNT3 (210%) and ENT2 (65%) mRNA. Abomasal infusion increased (54%) ileal expression of ENT2 mRNA and tended to increase (50%) jejunal ENT2 mRNA expression. This study has uniquely characterized the pattern of NT mRNA expression by growing beef cattle and found that the mRNA abundance for CNT3, ENT1, and ENT2 in small intestinal epithelia can be increased by increasing the luminal supply of nucleotides (CNT3, ENT1, ENT2) or glucose (ENT2).

摘要

在反刍动物中,微生物来源的核酸是氮的主要来源,并以核苷形式被小肠上皮吸收。尽管已经描述了牛的两种核苷转运系统的生化活性,但对其基因表达的调控知之甚少。本研究旨在验证两个假设:(1)肉牛的小肠上皮细胞对3种浓缩型(CNT1、2、3)和2种平衡型(ENT1、2)核苷转运体(NT)的mRNA表达存在差异;(2)这些NT的表达对瘤胃来源微生物(即核苷)、能量(玉米淀粉水解物,SH)或两者的小肠腔供应有反应。18头安装了瘤胃和皱胃导管的安格斯阉牛(体重260±17 kg)按照1.33倍维持净能(NE(m))需求饲喂基于苜蓿块的日粮。在3个时期内,每个时期的6头阉牛按体重(重与轻)进行分组。在每个组内,3头阉牛被随机分配到3种处理(n = 6):瘤胃和皱胃灌注水(对照)、瘤胃灌注SH/皱胃灌注水,或瘤胃灌注水/皱胃灌注SH。SH灌注剂量相当于代谢能摄入量的20%。在14或16天的灌注期后,屠宰阉牛,采集十二指肠、空肠和回肠上皮用于提取总RNA,并使用实时RT-PCR定量方法测定表达的mRNA相对量。所有5种NT mRNA均在每个上皮中表达,但它们的丰度在不同上皮之间存在差异。具体而言,所有5种NT mRNA的空肠表达均高于回肠,而CNT1、CNT3和ENT1 mRNA的空肠表达高于或趋于高于十二指肠表达。CNT2、CNT3和ENT2 mRNA的十二指肠表达高于回肠表达。关于SH灌注处理,瘤胃灌注增加了十二指肠中CNT3(67%)、ENT1(51%)和ENT2(39%)mRNA的表达以及回肠中CNT3(210%)和ENT2(65%)mRNA的表达。皱胃灌注增加了回肠中ENT2 mRNA的表达(54%),并趋于增加空肠中ENT2 mRNA的表达(50%)。本研究独特地描述了生长肉牛NT mRNA的表达模式,并发现通过增加核苷酸(CNT3、ENT1、ENT2)或葡萄糖(ENT2)的肠腔供应,可以增加小肠上皮中CNT3、ENT1和ENT2的mRNA丰度。

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