BMP4 knockdown of NCSCs leads to aganglionosis in the middle embryonic stage

Abstract

Transplacental bone morphogenetic protein (BMP)4 RNA interference (RNAi) is a technique used to knockdown genes in embryos. BMP4 are essential for the development of nervous system in the differentiation of neural crest stem cells (NCSCs). The failure of differentiation and migration of NCSCs may lead to aganglionosis. In the present study, pregnant mice were divided into three groups: Ringer's group, pSES group and RNAi‑BMP4 group. In order to silence the BMP4 gene in the first generation (F1), 11.5 day pregnant mice were injected with the small interfering RNA BMP4 plasmid, pSES or Ringer's solution via the tail vein. Semi‑quantitative reverse transcriptase‑polymerase chain reaction (RT‑PCR)and western blotting were employed to ensure the downregulation of BMP4. Finally, X‑rays were performed following a barium enema. Aganglionosis was diagnosed by general anatomy and immunohistochemistry. Compared with the control group, transplacental RNAi was able to downregulate the BMP4‑Smad4 of 11.5 day embryos, as determined by semi‑quantitative RT‑PCR and western blotting. The megacolons of the mice were demonstrated by X‑ray and confirmed by general anatomy. Aganglionosis of colonic mucosa and submucosa were diagnosed by pathology, and immunohistochemistry. Knockdown of BMP4 in pregnant mice at the middle embryonic stage led to aganglionosis. It was therefore demonstrated that BMP‑Smad was essential to the NCSCs of middle stage embryos. BMP‑Smad served important roles in the generation of aganglionosis. This technique of knockdown BMP4 gene may be used to establish an aganglionosis mouse model.

Functional Diversification of SRSF Protein Kinase to Control Ubiquitin-Dependent Neurodevelopmental Signaling

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Comprehensive Transcriptome Analysis of mRNA Expression Patterns of Early Embryo Development in Goat under Hypoxic and Normoxic Conditions

“…Functional enrichment analysis demonstrated that these DEGs were mainly related to biological processes and function regulation. For example, CDC25B, CCNA1, WEE1, PLK2, and CKS1B are related to cell cycle [ 48 , 49 , 50 , 51 , 52 ], while HSPB1, ZBTB33, BMP4, and PDGFA are involved in regulating embryonic growth and development [ 53 , 54 , 55 , 56 ].…”

Deciphering the molecular pathogenesis behind neurological manifestations of SARS-CoV-2 and drug repurposing, a systems biology approach

“…The most critical genes included six up-regulated shared genes (IL6, TNF, HOXA5, POU2F2, ITGB3, and STAT1) and 12 down-regulated shared genes (YY1, E2F6, ESR1, FOXO3, FOXO1, MEF2A, ATF3, ATF4, DDIT3, TCF4, BCL2L2, and BMP4). Several other studies have previously reported the mentioned critical genes to be involved in neural proliferation and differentiation (neurodevelopment), neurotransmission, synaptic plasticity, and myelination (Each molecule is separately described and referred to in Table 9) (Yang, Lindholm et al 2002, Nakanishi, Niidome et al 2007, Ragel, Couldwell et al 2007, He and Casaccia-Bonne l 2008, Imamura, Satoh et al 2008, Lange, Chavez et al 2008, Islam, Gong et al 2009 Raivich et al 2012, Pozo, Cingolani et al 2012, Cosker, Pazyra-Murphy et al 2013, Wang, Choi et al 2013, Ma, Tang et al 2014, Mazalouskas, Jessen et al 2015, Varney, Polston et al 2015, Doan, Kinyua et al 2016, Kennedy, Rahn et al 2016, Chen, Gao et al 2017, Lizen, Moens et al 2017, Higashi, Tanaka et al 2018, Li, Jin et al 2018, Liu, Amar et al 2018, Zhu, Carmichael et al 2018, Liu, Yu et al 2019, Majidi, Reddy et al 2019, Masgutova, Harris et al 2019, Wu and Donohoe 2019, Hartman and Czyz 2020, Pennycook, Vesela et al 2020. Below, we have hypothesized how some of these in-silico identi ed critical genes can play roles in neural manifestations of COVID-19 pathogenesis.…”