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技术说明:牛输卵管和子宫内膜芯片版本1:研究牛子宫内膜生物学和病理生理学的定制工具。

Technical note: Bovine oviduct and endometrium array version 1: a tailored tool for studying bovine endometrium biology and pathophysiology.

作者信息

Bauersachs S, Mitko K, Blum H, Wolf E

机构信息

Institute of Molecular Animal Breeding and Biotechnology, Ludwig-Maximilians University Munich, D-81377 Munich, Germany.

出版信息

J Dairy Sci. 2007 Sep;90(9):4420-3. doi: 10.3168/jds.2007-0132.

Abstract

Fertility problems are the main reason for slaughter of high-performance milk cows, because elongated calving intervals result in financial losses for the farmer and retard genetic progress. Genetic improvement of fertility would be of great benefit, but functional traits for effective selection are missing. Recent advances in functional genomics tools like DNA microarrays could be the key to identify gene expression patterns in the endometrium that correlate with maternal fertility. Therefore, a first version of a bovine oviduct and endometrium cDNA array was established that contains a set of 1,440 cDNA clones and long oligonucleotides representing 950 different genes. The major part of these genes has been identified in a series of differential gene expression studies in endometrium (different stages of the estrous cycle, d 18 pregnant vs. nonpregnant) and in oviduct epithelial cells (different stages of the estrous cycle) using a combination of subtracted cDNA libraries and cDNA array hybridization. Furthermore, cDNA clones of genes, which showed no changes in their mRNA levels in the analyzed tissues, were added as controls. Reproducibility of the array hybridization, a comparison with the Affymetrix bovine genome array, and confirmation of differential gene expression with reverse transcription-quantitative PCR is shown. Potential future applications include systematic studies of interactions between metabolic status and functionality of the endometrium to identify genes that could be used for differential diagnosis of fertility problems. Further, endometrium transcriptome profiles may serve as novel traits to improve fertility by genetic selection.

摘要

繁殖问题是高性能奶牛被屠宰的主要原因,因为产犊间隔延长会给养殖户带来经济损失,并阻碍遗传进展。繁殖力的遗传改良将大有裨益,但缺乏用于有效选择的功能性状。DNA微阵列等功能基因组学工具的最新进展可能是识别子宫内膜中与母畜繁殖力相关的基因表达模式的关键。因此,构建了牛输卵管和子宫内膜cDNA阵列的第一版,其中包含一组1440个cDNA克隆和代表950个不同基因的长寡核苷酸。这些基因中的大部分已在一系列子宫内膜(发情周期的不同阶段、妊娠第18天的妊娠与未妊娠)和输卵管上皮细胞(发情周期的不同阶段)的差异基因表达研究中得到鉴定,研究采用了消减cDNA文库和cDNA阵列杂交相结合的方法。此外,还添加了在所分析组织中mRNA水平无变化的基因的cDNA克隆作为对照。展示了阵列杂交的可重复性、与Affymetrix牛基因组阵列的比较以及用逆转录定量PCR对差异基因表达的验证。未来潜在的应用包括对子宫内膜代谢状态与功能之间相互作用的系统研究,以识别可用于繁殖问题鉴别诊断的基因。此外,子宫内膜转录组图谱可作为通过遗传选择提高繁殖力的新性状。

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