Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

High osmotic pressure forPneumocystic carinii London Resin White embedding enables fine immunocytochemistry studies: I. Golgi complex and cell-wall synthesis

  • 35 Accesses

  • 6 Citations

Abstract

A method for embeddingPneumocystis carinii in hydrophilic resin (London Resin White) has been developed for immunocytochemistry studies. Using high osmotic pressure (about 850 mosmol) from fixation to embedding, this method improved the preservation of the fine structure as well as the antigenicity of rabbit-and SCID mouse-derivedP. carinii. Cytochemistry studies were performed using colloidal gold-conjugated lectins (concanavalin A, glycine max,Ulex europaeus) that reacted with the cytoplasmic components (endoplasmic reticulum, Golgi vesicles). Colloidal gold-conjugated streptavidin was also tested and was found to be reactive with the parasite cell wall and cytoplasmic components, which precludes its indiscriminate use inP. carinii immunocytochemistry studies.

This is a preview of subscription content, log in to check access.

References

  1. Alon R, Bayer E, Wilcheck M (1990) Streptavidin contains an RYD sequence which mimics the RGD receptor domain of fibronectin. Biochem Biophys Res Commun 170:1236–1241

  2. Blumenfeld W, Mandrell R, Jarvis G, McLeod Griffiss J (1990) Localization of host immunoglobulin G to the surface ofPneumocystis carinii. Infect Immun 58:456–463

  3. Cushion M, De Stephano J, Walzer P (1988)Pneumocystis carinii: surface reactive carbohydrates detected by lectin probes. Exp Parasitol 67:137–147

  4. De Stephano J, Cushion M, Sleight R, Walzer P (1990) Analysis ofPneumocystis carinii cyst wall: I. Evidence for an outer surface membrane. J Protozool 37:428–435

  5. Kaneshiro E, Cushion M, Walzer P, Jayasimhulu K (1989) Analysis ofPneumocystis carinii fatty acids. J Protozool 36:69–72

  6. Pallnault F, Dei-Cas E, Slomianny C, Soulez B, Camus D (1990) Golgi complex and lysosomes in rabbit-derivedPneumocystis carinii. Biol Cell 70:73–82

  7. Palluault F, Slomianny C, Soulerz B, Dei-Cas E, Camus D (1992) High osmotic pressure enables fine ultrastructural and cytochemical studies onPneumocystis carinii. I Epon embedding. Parasitol Res (in press)

  8. Pesanti E, Shanley J (1988) Glycoproteins ofPneumocystis carinii: characterization by electrophoresis and microscopy. J Infect Dis 158:1353–1359

  9. Roth J, Taatjes D, Tokuyasu K (1990) Contrasting of Lowicryl K4M thin sections. Histochemistry 95:123–136

  10. Soulez B, Dei-Cas E, Charet P, Mougeot G, Cailaux M, Camus D (1989) The young rabbit: a nonimmunosuppressed model forPneumocystis carinii pneumonia. J Infect. Dis 160:355–356

  11. Soulez B, Palluault F, Cesbron J, Dei-Cas E, Capron A, Camus D (1992) Introduction ofPneumocystis carinii, in a colony of SCID mice. J Protozool (in press)

  12. Stryer L (1981) Biochemistry, 2nd edn. W.H. Freeman and Company, San Francisco

  13. Walzer P (1991) Immunopathogenesis ofPneumocystis carinii infection. J Lab Clin 118:206–216

  14. Weber W, Ohlendorf D, Wendoloski J, Salemme F (1989) Structural origins of high affinity of biotin binding to streptavidin. Science 243:85–88

  15. Yoshikawa H, Tegoshi T, Yoshida Y (1987) Detection of carbohydrates onPneumocystis carinii by fluorescein-conjugated lectins. Parasitol Res 74:43–49

  16. Yoshikawa H, Morioka H, Yoshida Y (1988) Ultrastructural detection of carbohydrates in the pellicle ofPneumocystis carinii. Parasitol Res 74:537–543

  17. Yoshikawa H, DeBault L, Graves D (1991) Ultrastructural localization of epitopes recognized by monoclonal antibodies produced to ratPneumocys is carinii. J Protozool 38:185–205

Download references

Author information

Correspondence to F. Palluault.

Additional information

This investigation was supported in part by the Agence Nationale de Recherche sur le SIDA (ANRS). The senior author (F.P.) was awarded a scholarship from the Conseil Régional Nord-Pas de Calais

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Palluault, F., Soulez, B., Slomianny, C. et al. High osmotic pressure forPneumocystic carinii London Resin White embedding enables fine immunocytochemistry studies: I. Golgi complex and cell-wall synthesis. Parasitol Res 78, 482–488 (1992). https://doi.org/10.1007/BF00931568

Download citation

Keywords

  • Cell Wall
  • Glycine
  • Endoplasmic Reticulum
  • Fine Structure
  • Osmotic Pressure