Hayashi Ken-Go, Hosoe Misa, Kizaki Keiichiro, Fujii Shiori, Kanahara Hiroko, Takahashi Toru, Sakumoto Ryosuke
Division of Animal Breeding and Reproduction Research, Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization, Tsukuba, 305-0901, Japan.
Division of Animal Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, Tsukuba, 305-8602, Japan.
Reprod Biol Endocrinol. 2017 Mar 23;15(1):20. doi: 10.1186/s12958-017-0237-6.
Repeat breeding directly affects reproductive efficiency in cattle due to an increase in services per conception and calving interval. This study aimed to investigate whether changes in endometrial gene expression profile are involved in repeat breeding in cows. Differential gene expression profiles of the endometrium were investigated during the mid-luteal phase of the estrous cycle between repeat breeder (RB) and non-RB cows using microarray analysis.
The caruncular (CAR) and intercaruncular (ICAR) endometrium of both ipsilateral and contralateral uterine horns to the corpus luteum were collected from RB (inseminated at least three times but not pregnant) and non-RB cows on Day 15 of the estrous cycle (4 cows/group). Global gene expression profiles of these endometrial samples were analyzed with a 15 K custom-made oligo-microarray for cattle. Immunohistochemistry was performed to investigate the cellular localization of proteins of three identified transcripts in the endometrium.
Microarray analysis revealed that 405 and 397 genes were differentially expressed in the CAR and ICAR of the ipsilateral uterine horn of RB, respectively when compared with non-RB cows. In the contralateral uterine horn, 443 and 257 differentially expressed genes were identified in the CAR and ICAR of RB, respectively when compared with non-RB cows. Gene ontology analysis revealed that genes involved in development and morphogenesis were mainly up-regulated in the CAR of RB cows. In the ICAR of both the ipsilateral and contralateral uterine horns, genes related to the metabolic process were predominantly enriched in the RB cows when compared with non-RB cows. In the analysis of the whole uterus (combining the data above four endometrial compartments), RB cows showed up-regulation of 37 genes including PRSS2, GSTA3 and PIPOX and down-regulation of 39 genes including CHGA, KRT35 and THBS4 when compared with non-RB cows. Immunohistochemistry revealed that CHGA, GSTA3 and PRSS2 proteins were localized in luminal and glandular epithelial cells and stroma of the endometrium.
The present study showed that endometrial gene expression profiles are different between RB and non-RB cows. The identified candidate endometrial genes and functions in each endometrial compartment may contribute to bovine reproductive performance.
重复配种由于每次受孕的配种次数增加和产犊间隔延长,直接影响奶牛的繁殖效率。本研究旨在调查子宫内膜基因表达谱的变化是否与奶牛的重复配种有关。利用微阵列分析,研究了发情周期黄体中期重复配种母牛(RB)和非RB母牛子宫内膜的差异基因表达谱。
在发情周期的第15天,从RB(至少输精3次但未怀孕)和非RB母牛的黄体同侧和对侧子宫角收集肉阜(CAR)和肉阜间(ICAR)子宫内膜(每组4头母牛)。使用定制的15K牛寡核苷酸微阵列分析这些子宫内膜样本的整体基因表达谱。进行免疫组织化学以研究三种鉴定出的转录本的蛋白质在子宫内膜中的细胞定位。
微阵列分析显示,与非RB母牛相比,RB同侧子宫角的CAR和ICAR中分别有405个和397个基因差异表达。在对侧子宫角,与非RB母牛相比,RB的CAR和ICAR中分别鉴定出443个和257个差异表达基因。基因本体分析显示,参与发育和形态发生的基因在RB母牛的CAR中主要上调。与非RB母牛相比,在同侧和对侧子宫角的ICAR中,与代谢过程相关的基因在RB母牛中显著富集。在整个子宫(结合上述四个子宫内膜区域的数据)分析中,与非RB母牛相比,RB母牛显示37个基因上调,包括PRSS2、GSTA3和PIPOX,39个基因下调,包括CHGA、KRT35和THBS4。免疫组织化学显示,CHGA、GSTA3和PRSS2蛋白定位于子宫内膜的腔上皮和腺上皮细胞及基质中。
本研究表明,RB和非RB母牛的子宫内膜基因表达谱不同。在每个子宫内膜区域鉴定出的候选子宫内膜基因及其功能可能有助于牛的繁殖性能。