Plasmodium berghei ookinetes induce nitric oxide production in Anopheles pseudopunctipennis midguts cultured in vitro.
The Anopheles pseudopunctipennis nitric oxide synthase gene (ApNOS) was identified and its partial sequence showed high homology with NOS from A. stephensi, A. gambiae (putative sequence), and Drosophila melanogaster. ApNOS was mainly expressed in male and female adult mosquitoes and was induced by a blood meal. Nitric oxide (NO) was produced by in vitro-cultured mosquito midguts inoculated by enema with Plasmodium berghei ookinetes, Saccharomyces cerevisiae, Gram-positive bacteria (Micrococcus luteus), but not with Gram-negative bacteria (Klebsiella pneumoniae, Escherichia coli or Serratia marcescens). Dihydroxyphenylalanine (L-DOPA) oxidation induced the generation of NO in midguts in vitro, and hydrogen peroxide generated during its oxidation induced ApNOS expression. P. berghei ookinetes exposed in vitro to L-DOPA and sodium nitroprusside (a NO generator) were killed. These observations demonstrate that reactive oxygen and nitrogen intermediates constitute a part of the cytotoxic arsenal employed by Anopheles mosquitoes against microbial pathogens and Plasmodium ookinetes.
Herrera-Ortíz
Antonia
A
Centro de Investigaciones Sobre Enfermedades Infecciosas, Instituto Nacional de Salud Pública, Av. Universidad 655, Col. Sta. María Ahuacatitlan, CP 62508 Cuernavaca, Morelos, México.
Lanz-Mendoza
Humberto
H
Martínez-Barnetche
Jesús
J
Hernández-Martínez
Salvador
S
Villarreal-Treviño
Cuauhtémoc
C
Aguilar-Marcelino
Liliana
L
Rodríguez
Mario H
MH
D000595:Amino Acid Sequence; D000818:Animals; D000852:Anopheles / Q000382:microbiology / Q000469:parasitology*; D004064:Digestive System / Q000201:enzymology / Q000382:microbiology; D005260:Female; D020869:Gene Expression Profiling; D006090:Gram-Negative Bacteria / Q000472:pathogenicity; D006094:Gram-Positive Bacteria / Q000472:pathogenicity; D007980:Levodopa / Q000502:physiology; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000096:biosynthesis* / Q000502:physiology; D019001:Nitric Oxide Synthase / Q000378:metabolism; D010962:Plasmodium berghei / Q000472:pathogenicity / Q000502:physiology*; D017382:Reactive Oxygen Species; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015053:Zygote / Q000502:physiology
D017382:Reactive Oxygen Species; D009569:Nitric Oxide; D007980:Levodopa; D019001:Nitric Oxide Synthase