Fertility and early embryonic development in a CD46-edited Gir heifer with reduced susceptibility to BVDV.

IF 3.1 2区 生物学 Q2 REPRODUCTIVE BIOLOGY Biology of Reproduction Pub Date : 2024-11-19 DOI:10.1093/biolre/ioae169
Alexandria P Snider, Aspen M Workman, Michael P Heaton, Brian L Vander Ley, Alexandria C Krueger, Tad S Sonstegard
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Abstract

Bovine viral diarrhea virus (BVDV) infection during pregnancy is a significant contributor to reproductive failures in cattle. The bovine receptor for BVDV (CD46) was previously edited with a six amino acid substitution (G82QVLAL to A82LPTFS) and shown to have significantly reduced BVDV susceptibility in a Gir heifer calf. Since a role for CD46 has been proposed in mammalian fertilization, our objective was to assess the edited heifer's fertilization rates, early embryonic development, and germline transmission conformation of the edit. Cumulus oocyte complexes were collected from the edited heifer and unedited females, fertilized with semen from an unedited bull and cultured until the blastocyst stage. Ultrasound examinations and serum progesterone concentration were also monitored to confirm estrous cyclicity in the CD46-edited heifer. Estrous cyclicity was normal with visualization of a corpus luteum and elevated progesterone concentrations. Fertilization rates and blastocyst development were not different in oocytes from edited and unedited controls. Genome sequence analysis of blastocysts confirmed germline transmission of either edited allele from the heifer. Subsequently, the CD46-edited heifer was artificially inseminated with semen from an unedited Gir bull and fertility status was confirmed with a diagnosed conception at day 35 of gestation. Thus, a six amino acid substitution in CD46 did not negatively affect fertilization of edited oocytes or early embryonic development when fertilized with semen from an unedited bull. An edited bull is still needed to similarly evaluate reproductive function of sperm cells carrying this CD46 edit.

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对 BVDV 易感性降低的 CD46 编辑 Gir 小母牛的生育能力和早期胚胎发育。
怀孕期间感染牛病毒性腹泻病毒(BVDV)是导致牛繁殖失败的一个重要原因。以前曾对牛的 BVDV 受体(CD46)进行过 6 个氨基酸的置换编辑(G82QVLAL 到 A82LPTFS),结果表明可显著降低 Gir 小母牛对 BVDV 的易感性。由于 CD46 在哺乳动物受精过程中的作用已被提出,我们的目标是评估编辑后小母牛的受精率、早期胚胎发育以及编辑后的种系传递构象。我们从编辑过的母牛和未编辑过的母牛身上采集了积层卵母细胞复合体,用未编辑过的公牛的精液进行受精,并培养至囊胚期。还监测了超声波检查和血清孕酮浓度,以确认 CD46 编辑母牛的发情周期。发情周期正常,可见黄体,孕酮浓度升高。已编辑和未编辑对照组卵母细胞的受精率和囊胚发育没有差异。囊胚的基因组序列分析证实,小母牛种系传播了其中一个编辑过的等位基因。随后,用未经编辑的 Gir 公牛的精液对 CD46 编辑过的小母牛进行人工授精,并在妊娠第 35 天确诊受孕。因此,CD46 中的六个氨基酸置换不会对编辑过的卵母细胞受精或与未编辑过的公牛精液受精时的早期胚胎发育产生负面影响。要对携带这种 CD46 编辑的精子细胞的生殖功能进行类似的评估,还需要一头经过编辑的公牛。
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来源期刊
Biology of Reproduction
Biology of Reproduction 生物-生殖生物学
CiteScore
6.30
自引率
5.60%
发文量
214
审稿时长
1 months
期刊介绍: Biology of Reproduction (BOR) is the official journal of the Society for the Study of Reproduction and publishes original research on a broad range of topics in the field of reproductive biology, as well as reviews on topics of current importance or controversy. BOR is consistently one of the most highly cited journals publishing original research in the field of reproductive biology.
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