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Copper-transporting ATPase is important for malaria parasite fertility.

Kenthirapalan S, Waters AP, Matuschewski K, Kooij TW - Mol. Microbiol. (2013)

Bottom Line: We show that male and female gametocytes are abundant in cutp(-) parasites, but activation of male microgametes and exflagellation were strongly impaired.A cross-fertilization assay demonstrated that female fertility was also severely abrogated.In conclusion, we provide experimental genetic and pharmacological evidence that a healthy copper homeostasis is critical to malaria parasite fertility of both genders of gametocyte and, hence, to transmission to the mosquito vector.

View Article: PubMed Central - PubMed

Affiliation: Max Planck Institute for Infection Biology, Parasitology Unit, 10117, Berlin, Germany.

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Related in: MedlinePlus

Cutp− parasites display reduced female fertility. Cross-fertilization assay to determine fertility of female and male cutp− gametes. The fertilization partners are p48/45− (pink), which produces only healthy female gametes and p47− (blue), which produces only healthy males. As positive and negative controls, a p47− × p48/45− cross-fertilization and homologous ookinete cultures were included. Ookinete cultures of cutp− parasites (black) yielded only 2% of the reference culture. A similar low level of ookinetes resulting from cross-fertilization with healthy male gametes (p47−) suggests a defect in female fertility of cutp− parasites. Partial rescue in cross-fertilization with healthy females (p48/45−) corroborates male gamete deficiency of cutp− parasites. Results were obtained from three independent experiments (repeated measures analysis of variance with Bonferroni's post-test; **P < 0.01; ns, non-significant).
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fig07: Cutp− parasites display reduced female fertility. Cross-fertilization assay to determine fertility of female and male cutp− gametes. The fertilization partners are p48/45− (pink), which produces only healthy female gametes and p47− (blue), which produces only healthy males. As positive and negative controls, a p47− × p48/45− cross-fertilization and homologous ookinete cultures were included. Ookinete cultures of cutp− parasites (black) yielded only 2% of the reference culture. A similar low level of ookinetes resulting from cross-fertilization with healthy male gametes (p47−) suggests a defect in female fertility of cutp− parasites. Partial rescue in cross-fertilization with healthy females (p48/45−) corroborates male gamete deficiency of cutp− parasites. Results were obtained from three independent experiments (repeated measures analysis of variance with Bonferroni's post-test; **P < 0.01; ns, non-significant).

Mentions: So far, we determined a ∼ 10-fold reduction in microgamete formation, yet observed a ∼ 100-fold reduced oocyst burden in cutp− parasites. To confirm a male fertility defect and test female fertility, we performed a cross-fertilization assay and quantified the resulting ookinetes, a functional read-out for gamete fertility (van Dijk et al., 2001; 2010,). As published previously, no ookinetes were detectable in cultures containing p48/45− or p47− parasites alone, whereas in co-cultures p47− male microgametes were able to productively fertilize p48/45− females and ookinetes were readily formed (Fig. 7). Next, we tested ookinete formation in cultures containing cutp− parasites alone. Ookinetes were observed only infrequently, approximating 2% of WT ookinete numbers (Fig. 7). This prominent reduction correlates with the very low oocyst formation (Fig. 4B), and apparently exceeds reduced exflagellation rates (Fig. 5A).


Copper-transporting ATPase is important for malaria parasite fertility.

Kenthirapalan S, Waters AP, Matuschewski K, Kooij TW - Mol. Microbiol. (2013)

Cutp− parasites display reduced female fertility. Cross-fertilization assay to determine fertility of female and male cutp− gametes. The fertilization partners are p48/45− (pink), which produces only healthy female gametes and p47− (blue), which produces only healthy males. As positive and negative controls, a p47− × p48/45− cross-fertilization and homologous ookinete cultures were included. Ookinete cultures of cutp− parasites (black) yielded only 2% of the reference culture. A similar low level of ookinetes resulting from cross-fertilization with healthy male gametes (p47−) suggests a defect in female fertility of cutp− parasites. Partial rescue in cross-fertilization with healthy females (p48/45−) corroborates male gamete deficiency of cutp− parasites. Results were obtained from three independent experiments (repeated measures analysis of variance with Bonferroni's post-test; **P < 0.01; ns, non-significant).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4016742&req=5

fig07: Cutp− parasites display reduced female fertility. Cross-fertilization assay to determine fertility of female and male cutp− gametes. The fertilization partners are p48/45− (pink), which produces only healthy female gametes and p47− (blue), which produces only healthy males. As positive and negative controls, a p47− × p48/45− cross-fertilization and homologous ookinete cultures were included. Ookinete cultures of cutp− parasites (black) yielded only 2% of the reference culture. A similar low level of ookinetes resulting from cross-fertilization with healthy male gametes (p47−) suggests a defect in female fertility of cutp− parasites. Partial rescue in cross-fertilization with healthy females (p48/45−) corroborates male gamete deficiency of cutp− parasites. Results were obtained from three independent experiments (repeated measures analysis of variance with Bonferroni's post-test; **P < 0.01; ns, non-significant).
Mentions: So far, we determined a ∼ 10-fold reduction in microgamete formation, yet observed a ∼ 100-fold reduced oocyst burden in cutp− parasites. To confirm a male fertility defect and test female fertility, we performed a cross-fertilization assay and quantified the resulting ookinetes, a functional read-out for gamete fertility (van Dijk et al., 2001; 2010,). As published previously, no ookinetes were detectable in cultures containing p48/45− or p47− parasites alone, whereas in co-cultures p47− male microgametes were able to productively fertilize p48/45− females and ookinetes were readily formed (Fig. 7). Next, we tested ookinete formation in cultures containing cutp− parasites alone. Ookinetes were observed only infrequently, approximating 2% of WT ookinete numbers (Fig. 7). This prominent reduction correlates with the very low oocyst formation (Fig. 4B), and apparently exceeds reduced exflagellation rates (Fig. 5A).

Bottom Line: We show that male and female gametocytes are abundant in cutp(-) parasites, but activation of male microgametes and exflagellation were strongly impaired.A cross-fertilization assay demonstrated that female fertility was also severely abrogated.In conclusion, we provide experimental genetic and pharmacological evidence that a healthy copper homeostasis is critical to malaria parasite fertility of both genders of gametocyte and, hence, to transmission to the mosquito vector.

View Article: PubMed Central - PubMed

Affiliation: Max Planck Institute for Infection Biology, Parasitology Unit, 10117, Berlin, Germany.

Show MeSH
Related in: MedlinePlus