a 2014

Cell motility in sporozoites of Cryptosporidium muris.

VALIGUROVÁ, Andrea, Isabelle FLORENT, Veronika MAZOUROVÁ and Janka MELICHEROVÁ

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

Language

English

Type of outcome

Konferenční abstrakt

Field of Study

10600 1.6 Biological sciences

Country of publisher

Sweden

Confidentiality degree

není předmětem státního či obchodního tajemství

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

Abstract

V originále

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.

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