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神经营养因子4促进孤雌激活后滋养外胚层谱系的特化并增强猪早期胚胎发育。

Neurotrophin-4 promotes the specification of trophectoderm lineage after parthenogenetic activation and enhances porcine early embryonic development.

作者信息

Kim Mirae, Lee Joohyeong, Cai Lian, Choi Hyerin, Oh Dongjin, Jawad Ali, Hyun Sang-Hwan

机构信息

Veterinary Medical Center and College of Veterinary Medicine, Laboratory of Veterinary Embryology and Biotechnology (VETEMBIO), Chungbuk National University, Cheongju, Republic of Korea.

Institute of Stem Cell and Regenerative Medicine (ISCRM), Chungbuk National University, Cheongju, Republic of Korea.

出版信息

Front Cell Dev Biol. 2023 Jul 13;11:1194596. doi: 10.3389/fcell.2023.1194596. eCollection 2023.

DOI:10.3389/fcell.2023.1194596
PMID:37519302
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10373506/
Abstract

Neurotrophin-4 (NT-4), a neurotrophic factor, appears to affect early embryonic development because it is secreted not only by neurons but also by oviductal and uterine epithelial cells. However, no studies have characterized the effects of NT-4 on early embryonic development in pigs. In this study, we applied the experimental model of parthenogenetic-activation (PA)-derived embryos. Herein, we investigated the effect of NT-4 supplementation during the culture (IVC) of embryos, analyzed the transcription levels of specific genes, and outlined the first cell lineage specification for porcine PA-derived blastocysts. We confirmed that NT-4 and its receptor proteins were localized in both the inner cell mass (ICM) and trophectoderm (TE) in porcine blastocysts. Across different concentrations (0, 1, 10, and 100 ng/mL) of NT-4 supplementation, the optimal concentration of NT-4 to improve the developmental competence of porcine parthenotes was 10 ng/mL. NT-4 supplementation during porcine IVC significantly ( < 0.05) increased the proportion of TE cells by inducing the transcription of TE lineage markers (, , and transcripts). NT-4 also reduced blastocyst apoptosis by regulating the transcription of apoptosis-related genes ( and transcripts) and improved blastocyst quality via the interaction of neurotrophin-, Hippo-yes-associated protein (Hippo-YAP) and mitogen-activated protein kinase/extracellular regulated kinase (MAPK/ERK) pathway. Additionally, NT-4 supplementation during IVC significantly ( < 0.05) increased transcript levels and significantly ( < 0.01) decreased transcript levels, respectively, in the porcine PA-derived blastocysts. We also confirmed through fluorescence intensity that the YAP1 protein was significantly ( < 0.001) increased in the NT-4-treated blastocysts compared with that in the control. NT-4 also promoted differentiation into the TE lineage rather than into the ICM lineage during porcine early embryonic development. In conclusion, 10 ng/mL NT-4 supplementation enhanced blastocyst quality by regulating the apoptosis- and TE lineage specification-related genes and interacting with neurotrophin-, Hippo-YAP-, and MAPK/ERK signaling pathway during porcine embryo development.

摘要

神经营养因子4（NT-4）是一种神经营养因子，似乎会影响胚胎早期发育，因为它不仅由神经元分泌，输卵管和子宫上皮细胞也会分泌。然而，尚无研究描述NT-4对猪胚胎早期发育的影响。在本研究中，我们应用了孤雌激活（PA）胚胎的实验模型。在此，我们研究了胚胎培养（IVC）期间补充NT-4的效果，分析了特定基因的转录水平，并概述了猪PA胚胎来源囊胚的首次细胞谱系特化。我们证实NT-4及其受体蛋白定位于猪囊胚的内细胞团（ICM）和滋养外胚层（TE）中。在补充不同浓度（0、1、10和100 ng/mL）的NT-4后，提高猪孤雌胚胎发育能力的最佳NT-4浓度为10 ng/mL。猪IVC期间补充NT-4通过诱导TE谱系标记物（、和转录本）的转录，显著（<0.05）增加了TE细胞的比例。NT-4还通过调节凋亡相关基因（和转录本）的转录减少了囊胚凋亡，并通过神经营养因子、Hippo-Yes相关蛋白（Hippo-YAP）和丝裂原活化蛋白激酶/细胞外调节激酶（MAPK/ERK）途径的相互作用提高了囊胚质量。此外，IVC期间补充NT-4在猪PA胚胎来源的囊胚中分别显著（<0.05）增加了转录本水平，并显著（<0.01）降低了转录本水平。我们还通过荧光强度证实，与对照组相比，NT-4处理的囊胚中YAP1蛋白显著（<0.001）增加。在猪早期胚胎发育过程中，NT-4还促进分化为TE谱系而非ICM谱系。总之，在猪胚胎发育过程中，补充10 ng/mL NT-4通过调节与凋亡和TE谱系特化相关的基因，并与神经营养因子、Hippo-YAP和MAPK/ERK信号通路相互作用，提高了囊胚质量。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cd/10373506/b1d12283f03b/fcell-11-1194596-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cd/10373506/87f7ef247cf8/fcell-11-1194596-g001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cd/10373506/b1d12283f03b/fcell-11-1194596-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cd/10373506/87f7ef247cf8/fcell-11-1194596-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cd/10373506/c8d6f13a143b/fcell-11-1194596-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cd/10373506/218220a5051b/fcell-11-1194596-g003.jpg
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