Three-Dimensional Investigations of Virus-Associated Structures in the Nuclei with White Spot Syndrome Virus (WSSV) Infection in Red Swamp Crayfish (Procambarus clarkii)

SIMPLE SUMMARY: Infections of white spot syndrome virus could be a lethal disease in shrimp culture systems. The 3D structures of the virus-associated structures were obtained through electron tomography. The relationships between the mature and immature particles were connected by the localizations of viral proteins. Our study provided new structural information on the virus through 3D, which extends the limited 2D knowledge of pathogen assembling in the white spot disease, further contributing to the development of a strategy for therapy or prevention. ABSTRACT: White spot syndrome virus (WSSV) has been reported to cause severe economic loss in the shrimp industry. With WSSV being a large virus still under investigation, the 3D structure of its assembly remains unclear. The current study was planned to clarify the 3D structures of WSSV infections in the cell nucleus of red swamp crayfish (Procambarus clarkii). The samples from various tissues were prepared on the seventh day post-infection. The serial sections of the intestinal tissue were obtained for electron tomography after the ultrastructural screening. After 3D reconstruction, the WSSV-associated structures were further visualized, and the expressions of viral proteins were confirmed with immuno-gold labeling. While the pairs of sheet-like structures with unknown functions were observed in the nucleus, the immature virions could be recognized by the core units of nucleocapsids on a piece of the envelope. The maturation of the particle could include the elongation of core units and the filling of empty nucleocapsids with electron-dense materials. Our observations may bring to light a possible order of WSSV maturation in the cell nucleus of the crayfish, while more investigations remain necessary to visualize the detailed viral–host interactions.