Immune stimulation and malaria infection impose reproductive costs in Anopheles gambiae via follicular apoptosis

Abstract

The employment of defense mechanisms is recognized as a costly life-history trait. In the malaria vector Anopheles gambiae, reproductive costs have been associated with both humoral and cellular innate immune responses and also with malaria infection. The resorption of developing oocytes associated with malaria infection is preceded by the programmed cell death, or apoptosis, of follicular cells. Here we demonstrate that apoptosis in ovarian follicular epithelial cells also occurs when mosquitoes are subjected to artificial immune-elicitors that induce a melanization response or humoral antimicrobial activity. Caspases are key cysteine proteases involved in apoptosis. Caspase-like activity was detected in epithelial cells in approximately 4.0% of the developing ovarian follicles of untreated, blood-fed, mosquitoes. Lipopolysaccharide injection resulted in a significant increase in anti-Micrococcus luteus humoral activity and a significant increase of 257.7% of follicles exhibiting apoptosis compared to results after saline injections. Melanization also triggered follicular apoptosis, which increased by 106.25% or 134.37% in Sephadex C-25 or G-25 bead-inoculated mosquitoes, respectively, compared to that in sham-injected ones. Ovaries from Plasmodium yoelii nigeriensis-infected mosquitoes exhibited a significant increase in follicular apoptosis of 440.9% compared to non-infected ones. Thus, at the time point investigated, infection had a much greater effect than artificial immune-elicitors. Death of follicular epithelial cells has been shown to lead to follicle resorption and hence a decrease in egg production. We propose the trade-off between reproductive fitness and immune defense in A. gambiae operates via the induction of apoptosis in ovarian follicles and that different immune responses impose costs via the same pathway.