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实验感染非细胞病变型牛病毒性腹泻病毒后体外培养牛胚胎中干扰素-α基因的上调。

Upregulation of interferon-alpha gene in bovine embryos produced in vitro in response to experimental infection with noncytophatic bovine-viral-diarrhea virus.

机构信息

Consejo Nacional de Investigaciones Científicas y Técnicas CONICET, Buenos Aires, Argentina.

Laboratorio de Virología Veterinaria, Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible INTA CONICET, Balcarce, Argentina.

出版信息

Mol Biol Rep. 2020 Dec;47(12):9959-9965. doi: 10.1007/s11033-020-05958-7. Epub 2020 Nov 23.

Abstract

In-vitro fertilization is a routine livestock-breeding technique widely used around the world. Several studies have reported the interaction of bovine viral-diarrhea virus (BVDV) with gametes and in-vitro-produced (IVP) bovine embryos. Since, gene expression in BVDV-infected IVP bovine embryos is scarcely addressed. The aim of this work was to evaluate the differential expression of genes involved in immune and inflammatory response. Groups of 20-25 embryos on Day 6 (morula stage) were exposed (infected) or not (control) to an NCP-BVDV strain in SOF medium. After 24 h, embryos that reached expanded blastocyst stage were washed. Total RNA of each embryo group was extracted to determine the transcription levels of 9 specific transcripts related with antiviral and inflammatory response by SYBR Green real time quantitative (RT-qPCR). Culture media and an aliquot of the last embryos wash on Day 7 were analyzed by titration and virus isolation, respectively. A conventional PCR confirmed BVDV presence in IVP embryos. A significantly higher expression of interferon-α was observed in blastocysts exposed to NCP-BVDV compared to the controls (p < 0.05). In this study, the upregulation of INFα and TLR7 genes involved in inflammatory and immune response in BVDV-infected IVP bovine embryos is a new finding in this field. This differential expression suggest that embryonic cells could function in a manner like immune cells by recognizing and responding early to interaction with viral pathogens. These results provide new insights into the action of BVDV on the complex molecular pathways controlling bovine early embryonic development.

摘要

体外受精是一种广泛应用于世界各地的常规家畜繁殖技术。几项研究报告了牛病毒性腹泻病毒(BVDV)与配子和体外生产(IVP)牛胚胎的相互作用。由于 BVDV 感染的 IVP 牛胚胎中的基因表达很少被研究。本工作的目的是评估参与免疫和炎症反应的基因的差异表达。第 6 天(桑葚胚阶段)的 20-25 个胚胎组暴露(感染)或不暴露(对照)于 SOF 培养基中的 NCP-BVDV 株。24 小时后,达到扩展囊胚阶段的胚胎被洗涤。从每组胚胎中提取总 RNA,通过 SYBR Green 实时定量(RT-qPCR)测定 9 种与抗病毒和炎症反应相关的特定转录本的转录水平。培养介质和第 7 天最后一批胚胎洗涤的等分试样分别通过滴定和病毒分离进行分析。常规 PCR 证实了 IVP 胚胎中存在 BVDV。与对照组相比,暴露于 NCP-BVDV 的囊胚中干扰素-α的表达显著升高(p<0.05)。在这项研究中,BVDV 感染的 IVP 牛胚胎中炎症和免疫反应相关的 INFα和 TLR7 基因的上调是该领域的一个新发现。这种差异表达表明胚胎细胞可以通过识别和早期响应与病毒病原体的相互作用,以类似于免疫细胞的方式发挥作用。这些结果为 BVDV 对控制牛早期胚胎发育的复杂分子途径的作用提供了新的见解。

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