Abstract
BackgroundOocysts of the malaria parasite form and develop in close proximity to the mosquito midgut basal lamina and it has been proposed that components of this structure play a crucial role in the development and maturation of oocysts that produce infective sporozoites. It is further suggested that oocysts incorporate basal lamina proteins into their capsule and that this provides them with a means to evade recognition by the mosquito's immune system. The site of production of basal lamina proteins in insects is controversial and it is still unclear whether haemocytes or midgut epithelial cells are the main source of components of the mosquito midgut basal lamina. Of the multiple molecules that compose the basal lamina, laminin is known to interact with a number of Plasmodium proteins. In this study, the localisation of mosquito laminin within the capsule and cytoplasm of Plasmodium berghei oocysts and in the midgut epithelial cells of Anopheles stephensi was investigated.ResultsAn ultrastructural examination of midgut sections from infected and uninfected An. stephensi was performed. Post-embedded immunogold labelling demonstrated the presence of laminin within the mosquito basal lamina. Laminin was also detected on the outer surface of the oocyst capsule, incorporated within the capsule and associated with sporozoites forming within the oocysts. Laminin was also found within cells of the midgut epithelium, providing support for the hypothesis that these cells contribute towards the formation of the midgut basal lamina.ConclusionWe suggest that ookinetes may become coated in laminin as they pass through the midgut epithelium. Thereafter, laminin secreted by midgut epithelial cells and/or haemocytes, binds to the outer surface of the oocyst capsule and that some passes through and is incorporated into the developing oocysts. The localisation of laminin on sporozoites was unexpected and the importance of this observation is less clear.
Highlights
Oocysts of the malaria parasite form and develop in close proximity to the mosquito midgut basal lamina and it has been proposed that components of this structure play a crucial role in the development and maturation of oocysts that produce infective sporozoites
We show by semi-quantitative immunogold labelling that, at 14 days post- infection, laminin coats and is incorporated into the capsule of P. berghei oocysts
Immunogold labelling for laminin was observed in sporozoites within maturing oocysts and we suggest a putative role for laminin in protecting both the oocyst and sporozoite from the mosquito defence system
Summary
Oocysts of the malaria parasite form and develop in close proximity to the mosquito midgut basal lamina and it has been proposed that components of this structure play a crucial role in the development and maturation of oocysts that produce infective sporozoites. The current emphasis on the development of new transmission blocking strategies to control malaria, and in particular the genetic manipulation of mosquitoes, make it essential to achieve a better understanding of the interactions between the vector and parasite. The ookinete penetrates the midgut epithelium at the apical junction of two epithelial cells and may transiently traverse several cells before exiting the basolateral membrane of the midgut epithelium. There it stops beneath the basal lamina (BL) and transforms via a took stage [5] into a sessile spherical oocyst [4]
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