ApiLoc - A database of published protein sub-cellular localisation in Apicomplexa

version 3 (curated until May 28, 2011)

In vitro induction of Neospora caninum bradyzoites in vero cells reveals differential antigen expression, localization, and host-cell recognition of tachyzoites and bradyzoites.

Vonlaufen, N., Guetg, N., Naguleswaran, A., Muller, N., Bjorkman, C., Schares, G., von Blumroeder, D., Ellis, J., Hemphill, A. (2004 Jan, Infect Immun)

We report on an optimized method for the in vitro culture of tissue cyst-forming Neospora caninum bradyzoites in Vero cells and the separation of viable parasites from host cells. Treatment of tachyzoite-infected Vero cell cultures with 17 microM sodium nitroprusside for 8 days severely scaled down parasite proliferation, led to reduced expression of tachyzoite surface antigens, and induced the expression of the bradyzoite marker NcBAG1 and the cyst wall antigen recognized by the monoclonal antibody MAbCC2. Transmission electron microscopy demonstrated that intracellular parasites were located within parasitophorous vacuoles that were surrounded by a cyst wall-like structure, and the dense granule antigens NcGRA1, NcGRA2, and NcGRA7 were incorporated into the cyst wall. Adhesion-invasion assays employing purified tachyzoites and bradyzoites showed that tachyzoites adhered to, and invaded, Vero cells with higher efficiency than bradyzoites. However, removal of terminal sialic acid residues from either the host cell or the parasite surface increased the invasion of Vero cells by bradyzoites, but not tachyzoites.

PubMed: 14688139, full text

Localisation information

NCLIV_036400 (GRA1) dense granule protein 1 / major antigenp24, putative

Experimental localisation: cyst wall during bradyzoite, posterior cytoplasm during tachyzoite, anterior cytoplasm during tachyzoite
  • Species: Neospora caninum
  • Quote inferring localisation: "Granular-type immunolabeling was found predominantly within the parasite cytoplasm at the anterior and posterior ends of the tachyzoites. In contrast, bradyzoites exhibited staining for NcGRA1 (see Fig. 4D to F) and NcGRA7 (see Fig. 4J to L) that had largely shifted toward the periphery of the vacuole. For NcGRA2 (see Fig. 4G to I), peripheral labeling was not as pronounced but was clearly evident as well."
  • Microscopy type: light, EM
  • Microscopy method: polylonal antibody
  • Strain:
  • Gene model mapping comments: blast from AAK16239, annotation matches, incorrect gene model?
  • Localisation record: cyst wall during bradyzoite, posterior cytoplasm and anterior cytoplasm during tachyzoite

NCLIV_045650 (GRA2) 28 kDa antigen, putative

Experimental localisation: cyst wall during bradyzoite, posterior cytoplasm during tachyzoite, anterior cytoplasm during tachyzoite
  • Species: Neospora caninum
  • Quote inferring localisation: "Granular-type immunolabeling was found predominantly within the parasite cytoplasm at the anterior and posterior ends of the tachyzoites. In contrast, bradyzoites exhibited staining for NcGRA1 (see Fig. 4D to F) and NcGRA7 (see Fig. 4J to L) that had largely shifted toward the periphery of the vacuole. For NcGRA2 (see Fig. 4G to I), peripheral labeling was not as pronounced but was clearly evident as well."
  • Microscopy type: light, EM
  • Microscopy method: polylonal antibody
  • Strain:
  • Gene model mapping comments: blast from AF196293, annotation matches
  • Localisation record: cyst wall during bradyzoite, posterior cytoplasm and anterior cytoplasm during tachyzoite
  • Comment: EM not shown in paper

NCLIV_021640 (GRA7, NCDG1) dense granule protein 7, putative

Experimental localisation: cyst wall during bradyzoite, posterior cytoplasm during tachyzoite, anterior cytoplasm during tachyzoite
  • Species: Neospora caninum
  • Quote inferring localisation: "Granular-type immunolabeling was found predominantly within the parasite cytoplasm at the anterior and posterior ends of the tachyzoites. In contrast, bradyzoites exhibited staining for NcGRA1 (see Fig. 4D to F) and NcGRA7 (see Fig. 4J to L) that had largely shifted toward the periphery of the vacuole. For NcGRA2 (see Fig. 4G to I), peripheral labeling was not as pronounced but was clearly evident as well."
  • Microscopy type: light, EM
  • Microscopy method: polylonal antibody
  • Strain:
  • Gene model mapping comments: blast from AF176649, annotation doesn't match
  • Localisation record: cyst wall during bradyzoite, posterior cytoplasm and anterior cytoplasm during tachyzoite
  • Comment: EM not shown in paper