Malarial development in mosquito and vaccine development. Mechanism of malarial sporozoite accumulation in the salivary glands of Anopheline mosquitoes.

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We studied the mechanism of malarial sporozoite accumulation of Plasmodium berghei in the salivery gland of Anopheles stephensi.1. We got following results with the observation of midgut infected with Plasmodium berghei by transmission electron microscope (TEM).(1) Ookinate reached the basal lamina of midgut through the cell junction 24 hours after blood was ingested. (2) Two days after infection, ookinate whose diameter was 6 um has stopped moving and became round. (3) Eight days after infection, there were many nuclei which developed into sporozoites inside the oocyst. (4) Fourteen days after infection, oocyst developed matured oocyst, and many sparozoite exsisted around the core.2. To observe the penetration of the sporozoites into salivary glands, we fixed living mosquitoes by 2.5% osmium and dissected them to remove the salivary glands after 2 hours. We got following results by scanning electron microscope and TEM.(1) Sporozoites exsisted on the surface and some of them were penetrating in the salivary glands. (2) Inside of the salivary glands, sporozoites entered the cytoplasm of the cell from outside and gathered in the secretory cavity.3. We extracted mRNA from salivery gland of mosquito and synthesized cDNA.The cDNA was ligated lambdagtll phage and cDNA libraries were produced. These were screened by the monoclonal antibody of salivery gland and 4 possitive clones were selected. The size of insert of 4 clones were 1450bp, 360bp, 220bp and 790bp.