title: Nitric oxide synthase in filariae: demonstration of nitric oxide production by embryos in Brugia malayi and Acanthocheilonema viteae.
abstract: The radical gas nitric oxide (NO) is synthesized by nitric oxide synthase (NOS) from l-arginine and molecular oxygen. Nitric oxide is an important signaling molecule in invertebrate and vertebrate systems. Previously we have shown that NOS is localized to more tissues in Brugia malayi than has been reported in Ascaris suum. In this paper, we analyze the distribution of NOS in Acanthocheilonema viteae, a filarial nematode that differs from B. malayi in that A. viteae females release microfilariae without a sheath. A. viteae is also one of a few filarial parasites without the Wolbachia intracellular endosymbiont. By use of a specific antibody, NOS was demonstrated in extracts of A. viteae and Dirofilaria immitis. The localization pattern of NOS in A. viteae was similar to that seen in B. malayi, with the enzyme localized to the body wall muscles of both sexes, developing spermatozoa, intrauterine sperm, and early embryos. By use of DAF-2, a fluorescent indicator specific for nitric oxide, the embryos of B. malayi and A. viteae were demonstrated to produce NO ex utero. The near identical staining patterns seen in A. viteae and B. malayi argue that NO is not produced by Wolbachia, nor is it produced by the nematodes in response to the infection. Localization of NOS to the sperm of filarial nematodes suggests a role for NO during fertilization as has been described for sea urchin and ascidian fertilization. Demonstration of the activity of embryonic NOS supports our earlier hypothesis that NO is a signaling molecule during embryogenesis in filarial nematodes.
authors.affiliation: Department of Biology, Tufts University, Medford, Massachusetts 02155, USA.
authors.forename: K M
authors.initials: KM
authors.lastname: Pfarr
authors.forename: S
authors.initials: S
authors.lastname: Qazi
authors.forename: J A
authors.initials: JA
authors.lastname: Fuhrman
mesh_terms: D000818:Animals; D017178:Brugia malayi / Q000196:embryology / Q000201:enzymology*; D004153:Dipetalonema / Q000196:embryology / Q000201:enzymology*; D005260:Female; D005849:Gerbillinae; D007150:Immunohistochemistry; D008297:Male; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D009569:Nitric Oxide; D019001:Nitric Oxide Synthase