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Background    Brugia malayi , like most human filarial parasite species, harbors an endosymbiotic bacterium of the genus  Wolbachia . Elimination of the endosymbiont leads to sterilization of the adult female. Previous biochemical and genetic studies have established that communication with its endobacterium is essential for survival of the worm.    Methodology/Principal findings  We used electron microscopy to examine the effects of antibiotic treatment on  Wolbachia  cell structure. We have also used microarray and quantitative RT-PCR analyses to examine the regulation of the  B. malayi  transcripts altered in response to the anti- Wolbachia  treatment. Microscopy of worms taken from animals treated with tetracycline for 14 and 21 days (14 d and 21 d) demonstrated substantial morphologic effects on the  Wolbachia  endobacterium by 14 d and complete degeneration of the endobacterial structures by 21 d. We observed upregulation of transcripts primarily encoding proteins involved in amino acid synthesis and protein translation, and downregulation of transcripts involved in cuticle biosynthesis after both 7 d and 14 d of treatment. In worms exposed to tetracycline in culture, substantial effects on endobacteria morphology were evident by day 3, and extensive death of the endobacteria was observed by day 5. In a detailed examination of the expression kinetics of selected signaling genes carried out on such cultured worms, a bimodal pattern of regulation was observed. The selected genes were upregulated during the early phase of antibiotic treatment and quickly downregulated in the following days. These same genes were upregulated once more at 6 days post-treatment.    Conclusions/Significance  Upregulation of protein translation and amino acid synthesis may indicate a generalized stress response induced in  B. malayi  due to a shortage of essential nutrients/factors that are otherwise supplied by  Wolbachia . Downregulation of transcripts involved in cuticle biosynthesis perhaps reflects a disruption in the normal embryogenic program. This is confirmed by the expression pattern of transcripts that may be representative of the worms' response to  Wolbachia  in different tissues; the early peak potentially reflects the effect of bacteria death on the embryogenic program while the second peak may be a manifestation of the adult worm response to the affected bacteria within the hypodermis.

Brugia malayi Gene Expression in Response to the Targeting of the Wolbachia Endosymbiont by Tetracycline Treatment