Performing Immunohistochemistry in Mosquito Salivary Glands.

Studying protein localization in mosquito salivary glands provides novel insights on the function and physiological relevance of salivary proteins and also provides an avenue to study interactions between mosquitoes and pathogens. Salivary proteins display compartmentalization. For example, proteins involved in blood feeding are stored in the medial and distal lateral lobes, whereas proteins related to sugar metabolism localize to the proximal portion of the lateral lobes. Immunohistochemistry assays use antibodies raised against recombinant salivary proteins to reveal the protein localization and interactions within the tissue. In this assay, permeabilization of the salivary glands allows the antibodies to enter the cells and bind their target proteins. The primary antibody-antigen complexes are later marked with fluorescently labeled secondary antibodies. Antibodies that recognize pathogen-specific proteins can also be incorporated in these assays, providing information about pathogen localization within the salivary glands or pathogen interactions with mosquito salivary proteins. Here, we introduce immunohistochemistry assays for use in mosquito salivary glands.