Spatial distribution of orally administered viral fusolin protein in the insect midgut and possible synergism between fusolin and digestive proteases to disrupt the midgut peritrophic matrix

Wataru Mitsuhashi, Sachiko Shimura, Kazuhisa Miyamoto, Takafumi N. Sugimoto

Oral inoculation of entomopoxvirus spindles, microstructures composed of fusolin protein, causes disruption of the peritrophic matrix (PM), a physical barrier against microbe infection, in the insect midgut. Although the atomic structure of fusolin has been determined, little has been directly elucidated of the mechanism of disruption of the PM. In the present study, we first performed an immunohistochemical examination to determine whether fusolin acts on the PM directly or indirectly in the midgut of Bombyx mori larvae that were inoculated with spindles of Anomala cuprea entomopoxvirus. This revealed that the PM, rather than the midgut cells, was the attachment site for fusolin. Fusolin broadly attached to the PM from the anterior to the posterior region, both to its ectoperitrophic and endoperitrophic surfaces and within the PM. These results likely explain why the whole of the PM is rapidly disintegrated. Second, we administered protease inhibitors mixed with spindles and observed decreased midgut protease activity and reduced disruption of the PM. This suggests that midgut protease(s) is also positively involved in PM disruption. Based on the present results, we propose an overall mechanism for the disruption of the PM by administration of fusolin.