Primary Neuron Culture Of Newborn Mice And Rats And Hantavirus Infection Of Neuron Culture In Vitro

Hantavirus belongs to Bunyaviridae family. Infections with certain hantavirus species can cause human disease as hantavirus pulmonary syndrome (HPS) and hemorrhagic fever with renal syndrome (HFRS). HFRS is caused by pathogenic Hantavirus species like Hantaan virus (HTNV), Seoul virus, Dobrava virus, and Puumala virus (PUUV). HTV infected abroad range of organs and cells in vivo, and brain is one of the sensitive target organs. Our previous work showed that HTV can lethally infect newborn suckling mice, the viral antigen and RNA can be detected in the cytoplasm of neurons in cerebral cortex and hippocampus, as well as Purkinje cell of cerebella cortex. To explore the direct effect of HTV infection on neuron, the primary cortical neuron cultures of newborn mice and rats, less than 24hr of age, was prepared for HTV L99 strain infection in vitro. After 1d, 2d, 3d, 4d, 5d, 6d, 7d, 8d, and 9d of virus infection, the cultures were harvested for immunohistochemical detection of virus nucleocapsid protein and morphological observation. The results showed that the HTV NP can be detected as early as 2d in neuron culture of newborn suckling mice and 1d in newborn suckling rats after infection, and localized in the cytoplasm with fine granular pattern. The positive rates and staining densities for NP were progressively and significantly increased in a time-dependent manner (P<0.01). The cytopathogenic effect can be found at 2d after infection, as demonstrated by...