Fig. 7: Sexual and exo-erythrocytic stage development of PbGSKO parasites.
From: Distinct evolution of type I glutamine synthetase in Plasmodium and its species-specific requirement
a Number of male and female gametocytes observed in Giemsa-stained smears of Balb/c mice (n = 6) prepared on day 8 post-infection. b Number of exflagellation centers observed in glutamine-free (Gln-) and glutamine-containing (Gln+) exflagellation medium for the blood collected on day 8 post-infection. n = 7 different Balb/c mice. c In vitro ookinete formation in glutamine-free (Gln-) and glutamine-containing (Gln+) medium. n = 3 different Balb/c mice. For a–c (mean ± SD; n.s - not significant, unpaired t-test; two-sided). d Giemsa-stained images for in vivo ookinetes observed in smears prepared from the blood bolus collected at 21 h post-fed mosquito guts. Images were captured using 100x objective. Scale bar = 5 μM. e In vivo ookinete formation in the mosquito guts dissected at 21 h post-feeding. f Mercurochrome staining for the day 10 post-fed mosquito guts. Black arrows indicate oocysts. Images were captured using 20x objective. Scale bar = 20 μM. g In vivo oocyst formation in the mosquito guts dissected on day 10 post-feeding. h Bright field images of salivary glands from day 16 post-fed mosquitoes. Black arrows indicate sporozoites. Scale bar = 20 μM. i In vivo sporozoite formation in the mosquito salivary glands dissected on day 17 post-feeding. For d–i n = 30 mosquitoes from three independent batches. (n.s - not significant, **P < 0.01, unpaired t-test; two-sided) j Growth curve analysis performed for Balb/c mice infected with sporozoites assessing the ability of PbGSKO sporozoites to undergo exo-erythrocytic stage development. Appearance of blood stage parasites was monitored by Giemsa smears prepared form peripheral blood. n = 3. (mean ± SD; n.s - not significant, Two-way ANOVA). k–m Immunofluorescence analysis of PbGSKO ookinete (k), sporozoite (l) and exo-erythrocytic stage (m) with PbGS antibodies. n = 2 independent experiments. Scale bar for ookinete and sporozoite = 5 μM. UIS4 antibody was used to identify the exo-erythrocytic stages. Scale bar for exo-erythrocytic stage = 20 μM. Images were captured using 60x/100x objective. Source data are provided as a Source Data file.
