P. falciparum gametocytes exhibit increased sensitivity to mitochondrial tricarboxylic acid (TCA)-cycle inhibitors. Ring-infected red blood cells (RBCs), gametocytes, and uninfected RBCs were cultured in the presence or absence of sodium fluoroacetate (NaFAc), and the metabolic and morphological effects were assessed. (A) After culturing in the presence or absence of 1 mmol/l NaFAc, metabolites were extracted from schizont-infected and uninfected RBCs (at 38 hours) and gametocytes (at 24 hours) and analyzed by gas chromatography–mass spectrometry (GC-MS). Bars represent abundance of metabolites in drug-treated cells compared with a drug-free control. Results are the mean of three to four biological replicates. Abbreviations: 3PGA, 3-phosphoglycerate; α-GP, α-glycerophosphate; α-KG, α-ketoglutarate; Ala, alanine; Asp, aspartate. Cit, citrate; F6P, fructose 6-phosphate; Fum, fumarate; G6P, glucose 6-phosphate; GABA, γ-aminobutyric acid; Glu, glutamate; Lac, lactic acid; Mal, malate; Pyr, pyruvate; R5P, ribose 5-phosphate; Ru5P, ribulose 5-phosphate; sH7P, sedo-heptulose 7-phosphate; Suc, succinate, (B) Gametocytes were cultured in standard culture medium with or without the addition of 1 or 10 mmol/l NaFAc. Stage distribution and parasitemia levels were assessed in smears made on days 0 to 7, and the percentage of fully mature (Stage V) gametocytes calculated (see Additional file 6 for representative smears). Error bars represent SEM, where n = 3. Day 7 gametocytes were labeled with (C) rhodamine-123 (Rho123) or (D) BODIPY-TR-ceramide. In untreated gametocytes, these dyes accumulated (white arrows) in (C) the reticulate mitochondrion and (D) intracellular membranes, but redistributed to the parasite plasma membrane in treated parasites (yellow arrows). Bright field (BF) images are shown. Scale bar = 3 μm.