miR-142-5p Disrupts Neuronal Morphogenesis Underlying Porcine Hemagglutinating Encephalomyelitis Virus Infection by Targeting Ulk1

Li, Zi; Lan, Yungang; Zhao, Kui; Lv, Xiaoling; Ding, Ning; Lu, Huijun; Zhang, Jing; Yue, Huiqing; Shi, Junchao; Song, Deguang; Gao, Feng; He, Wenqi

MicroRNAs are involved in post-transcriptional regulation, including development, differentiation, apoptosis, proliferation, and other processes (Bartel, 2009; Shukla et al., 2011) . Over the past decade, numerous insights into the roles of microRNAs have greatly changed our conceptualization of the virus/host relationship (Friedman et al., 2009; Feldman and Tibbetts, 2015; Mizuguchi et al., 2015 Mizuguchi et al., , 2016 Ruiz and Russell, 2015) . For example, miR-27 was found to be degraded in latent Herpesvirus saimiri (HVS) infection, and as a destroyer caused FIGURE 9 | miR-142-5p antagomir blocked PHEV proliferation in mice. (A) Design and optimization of the appropriate treatment strategy for PHEV-infected mice. qRT-PCR showed that endogenous miR-142-5p was specially inhibited by antagomirs, and that Ulk1 expression was significantly enhanced. (B) The freshly isolated cerebral cortex lysate was harvested for Western blot at 6 dpi, and Ulk1 protein was enhanced after miR-142-5p treatment. (C) Mice were monitored daily for clinical signs, and survival curves are presented per time point. n =10 mice per group for three independent experiments. (D) qRT-PCR of PHEV mRNA showed viral replication clearly blocked in the antagomir-treated mice. (E) Visualization of PHEV-infected cerebral cortexes and hippocampi from mice at 5 dpi. Upper panels show the mice incubated with PHEV alone, middle panels represent the antagomir-treated mice after PHEV infection and the lower panels represent the healthy mice. (F) Quantitative analyses of the PHEV staining intensity averaged (AFU) over the area showed statistically attenuated signals in the antagomir-treated mice. Error bars for all experiments represent the geometric means ± s.d. Asterisks indicate differences that were statistically significant (*P < 0.05, **P < 0.01, ***P < 0.001).