Myotropic effects of FMRFamide containing peptides on the heart of the mosquito Anopheles gambiae

• The Anopheles gambiae FMRFamide gene is alternatively spliced and encodes eight putative peptides.
• Peak transcription of FMRFamide and its putative receptor occurs in second instar larvae and around eclosion.
• In adult mosquitoes, FMRFamide and its putative receptor are primarily transcribed in the thorax and head.
• Exposure to 1 × 10−6 M of FMRFamide or SALDKNFMRFamide increases mosquito heart rates.
• Exposure to 1 × 10−2 M of FMRFamide or SALDKNFMRFamide decreases mosquito heart rates.

FMRFamide-like peptides (FLPs) are produced by invertebrate and vertebrate animals, and regulate diverse physiological processes. In insects, several FLPs modulate heart physiology, with some increasing and others decreasing dorsal vessel contraction dynamics. Here, we describe the FMRFamide gene structure in the mosquito, Anopheles gambiae, quantify the developmental and spatial expression of FMRFamide and its putative receptor (FMRFamideR), and show that the peptides FMRFamide and SALDKNFMRFamide have complex myotropic properties. RACE sequencing showed that the FMRFamide gene encodes eight putative FLPs and is alternatively spliced. Of the eight FLPs, only one is shared by A. gambiae, Aedes aegypti and Culex quinquefasciatus: SALDKNFMRFamide. Quantitative PCR showed that peak expression of FMRFamide and FMRFamideR occurs in second instar larvae and around eclosion. In adults, FMRFamide is primarily transcribed in the head and thorax, and FMRFamideR is primarily transcribed in the thorax. Intravital video imaging of mosquitoes injected FMRFamide and SALDKNFMRFamide revealed that at low doses these peptides increase heart contraction rates. At high doses, however, these peptides decrease heart contraction rates and alter the proportional directionality of heart contractions. Taken altogether, these data describe the FMRFamide gene in A. gambiae, and show that FLPs are complex modulators of mosquito circulatory physiology.

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