Cell motility in sporozoites of Cryptosporidium muris.

VALIGUROVÁ, Andrea, Isabelle FLORENT, Veronika MAZOUROVÁ and Janka MELICHEROVÁ. Cell motility in sporozoites of Cryptosporidium muris. In The 5th. International Giardia and Cryptosporidium Conference 2014. 2014.

Basic information

• Original name: Cell motility in sporozoites of Cryptosporidium muris.
• Authors: VALIGUROVÁ, Andrea (703 Slovakia, guarantor, belonging to the institution), Isabelle FLORENT (250 France), Veronika MAZOUROVÁ (203 Czech Republic) and Janka MELICHEROVÁ (703 Slovakia).
• Edition: The 5th. International Giardia and Cryptosporidium Conference 2014, 2014.

Other information

• Original language: English
• Type of outcome: Conference abstract
• Field of Study: 10600 1.6 Biological sciences
• Country of publisher: Sweden
• Confidentiality degree: is not subject to a state or trade secret
• RIV identification code: RIV/00216224:14310/14:00073932
• Organization unit: Faculty of Science
• Keywords (in Czech): Apicomplexa; Cryptosporidium; motility; sporozoite; actin; mysoin; tubulin
• Keywords in English: Apicomplexa; Cryptosporidium; motility; sporozoite; actin; mysoin; tubulin
• Tags: Actin, Apicomplexa, Cryptosporidium, motility, myosin, sporozoite, tubulin

Abstract

Although the motility of cryptosporidian sporozoites is considered as the main mechanism facilitating the host cell invasion, our observations show that motility of C. muris sporozoites is very limited and featureless, and differs from other apicomplexan zoites. As cryptosporidian sporozoites possess a single rhoptry, they have only one attempt for successful attachment to the host cell. Within the host organism, released sporozoites of C. muris rapidly penetrate deeply into the bottom of the pits of the gastric glands to avoid the adverse conditions in the host stomach. In cell cultures, using various media, including those enriched by BSA and vitamins, the activity of freshly released sporozoites decreases very rapidly and after several minutes, sporozoites do not show any signs of vitality. The apical region of invasive sporozoites is obviously prolonged and their three-layered pellicle is smooth lacking any grooves or folds. Using immunofluorescence we were able to obtain myosin labelling, which is considered an essential part of apicomplexan motility motor. Labelling of actin with a specific antibody recognizing the actin in Toxoplasma gondii and Plasmodium falciparum was not successful despite multiple repetitions of staining procedure of C. muris sporozoites. However, immunoblotting assays of sporozoites soluble proteins indicated the presence of actin (42 kDa) in relatively low concentrations. In addition, we were able to amplify by PCR and sequence the C. muris actin gene from genomic DNA. Using the both immunofluorescence and immunoblotting, we were able to detect alfa-tubulin (50 kDa), which represents an elemental component of subpellicular microtubules. Therefore, the currently obtained data support the presence, in C. muris, of basic mechanism of apicomplexan motility that is expected to be based on the orientation of the actomyosin motor by subpellicular microtubules.

Abstract (in Czech)

Although the motility of cryptosporidian sporozoites is considered as the main mechanism facilitating the host cell invasion, our observations show that motility of C. muris sporozoites is very limited and featureless, and differs from other apicomplexan zoites. As cryptosporidian sporozoites possess a single rhoptry, they have only one attempt for successful attachment to the host cell. Within the host organism, released sporozoites of C. muris rapidly penetrate deeply into the bottom of the pits of the gastric glands to avoid the adverse conditions in the host stomach. In cell cultures, using various media, including those enriched by BSA and vitamins, the activity of freshly released sporozoites decreases very rapidly and after several minutes, sporozoites do not show any signs of vitality. The apical region of invasive sporozoites is obviously prolonged and their three-layered pellicle is smooth lacking any grooves or folds. Using immunofluorescence we were able to obtain myosin labelling, which is considered an essential part of apicomplexan motility motor. Labelling of actin with a specific antibody recognizing the actin in Toxoplasma gondii and Plasmodium falciparum was not successful despite multiple repetitions of staining procedure of C. muris sporozoites. However, immunoblotting assays of sporozoites soluble proteins indicated the presence of actin (42 kDa) in relatively low concentrations. In addition, we were able to amplify by PCR and sequence the C. muris actin gene from genomic DNA. Using the both immunofluorescence and immunoblotting, we were able to detect alfa-tubulin (50 kDa), which represents an elemental component of subpellicular microtubules. Therefore, the currently obtained data support the presence, in C. muris, of basic mechanism of apicomplexan motility that is expected to be based on the orientation of the actomyosin motor by subpellicular microtubules.

Links

• GAP505/11/1163, research and development project: Name: Protizánětlivá aktivita extraktů z rostlin Indonésie a jejich účinek na průběh oportunních parazitóz

Investor: Czech Science Foundation

• GPP506/10/P372, research and development project: Name: Srovnávací morfologie a imunohistochemie v hodnocení fylogenetických vztahů mezi zástupci raných linií kmene Apicomplexa

Investor: Czech Science Foundation