Related Experiment Videos
Immunocytochemical identification of spore proteins in two microsporidia, with emphasis on extrusion apparatus
F Delbac1, F Duffieux, D David
1LBCP, UPESA CNRS 6023, Université Blaise Pascal, Aubière, France.
Abstract:
Microsporidia can form small spores with a unique invasive apparatus featuring a long polar tube whose extrusion allows entry of infectious sporoplasm into a host cell. The reactivity of mouse polyclonal antibodies raised against sporal proteins from two microsporidian species belonging to different genera (Glugea atherinae and Encephalitozoon cuniculi) was studied by western blotting and indirect immunofluorescence. Whole protein antisera provided a few cross-reactions relatable to some proteins of the spore envelope or polar tube. Ultrastructural immunocytochemistry with murine antibodies against protein bands separated by sodium dodecylsulphate polyacrylamide gel electrophoresis allowed the assignment of several proteins to the polar tube (34, 75 and 170 kDa in Glugea, 35, 55 and 150 kDa in Encephalitozoon). Antigenic similarities were detected for the Glugea 34 kDa and Encephalitozoon 35 kDa polar tube proteins. Species-specific proteins were shown to be located in either the lamellar polaroplast of Glugea or the spore envelope of Encephalitozoon.
Insights
This study characterized microsporidian spore proteins using mouse antibodies. Similarities were found in polar tube proteins between Glugea and Encephalitozoon species.
Area of Science:
- * Parasitology
- * Molecular Biology
- * Immunology
Background:
- * Microsporidia are obligate intracellular parasites forming spores with a unique polar tube for host cell invasion.
- * Understanding the proteins involved in spore structure and function is crucial for parasite biology.
- * Mouse polyclonal antibodies were generated against sporal proteins of Glugea atherinae and Encephalitozoon cuniculi.
Purpose of the Study:
- * To investigate the antigenic similarities and differences of sporal proteins between two distinct microsporidian genera.
- * To identify and localize specific proteins within the spore's polar tube and envelope using immunocytochemistry.
- * To characterize the reactivity of antibodies raised against whole sporal proteins.
Main Methods:
- * Western blotting and indirect immunofluorescence assays were employed to assess antibody reactivity.
- * Ultrastructural immunocytochemistry was used to localize proteins within the microsporidian spores.
- * Sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) was utilized to separate protein bands.
Main Results:
- * Whole protein antisera showed cross-reactions with spore envelope and polar tube proteins.
- * Specific polar tube proteins were identified: 34, 75, and 170 kDa in Glugea; 35, 55, and 150 kDa in Encephalitozoon.
- * Antigenic similarity was observed between Glugea 34 kDa and Encephalitozoon 35 kDa polar tube proteins.
Conclusions:
- * Mouse antibodies effectively identified conserved and species-specific proteins in microsporidian spores.
- * Key polar tube proteins exhibit antigenic similarities between different microsporidian genera.
- * Species-specific proteins are localized in the polaroplast (Glugea) and spore envelope (Encephalitozoon).