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A study of the cell biology of motility in Eimeria tenella sporozoites

Bruce, David Robert; (2000) A study of the cell biology of motility in Eimeria tenella sporozoites. Doctoral thesis (Ph.D), UCL (University College London). Green open access

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Abstract

Eimeria tenella is an obligate intracellular parasite within the phylum Apicomplexa. It is the causative agent of coccidiosis in domesticated chickens and under modem farming conditions can have a considerable economic impact. Motility is employed by the sporozoite to effect release from the sporocyst and enable invasion of appropriate host cells and occurs at an average speed of 16.7 6 ms-1. Frame by frame video analysis of gliding motility shows it to be an erratic non-substrate specific process and this observation was confirmed by studies of bead translocation across the cell surface occurring at an average speed of 16.9 7.6 ums-1. Incubation with cytochalasin D, 2,3-butanedione monoxime and colchicine, known inhibitors of the motility associated proteins actin, myosin and tubulin respectively, indicated that it is an actomyosin complex which generates the force to power sporozoite motility. Western blotting analysis confirmed the presence of actin with an apparent molecular weight of 43kDa and an unconventional myosin with an apparent molecular weight of 93kDa. Antibodies against the actin binding proteins spectrin, vinculin, filamin, -actinin, cofilin and tropomyosin failed to recognise any polypeptides in whole cell extracts. Immunofluoresence studies showed actin was found predominantly in the anterior third of the sporozoite. Myosin appeared to have a more widespread distribution, with a strong signal found at the margins of the cell. Genomic DNA samples were prepared and two degenerate primers against highly conserved regions of the myosin head were used in a polymerase chain reaction (PCR) to probe for the presence of myosin genes. These PCR products were inserted into suitable plasmids followed by amplification in bacteria. Selection of appropriate bacterial colonies and subsequent DNA sequence analysis identified a clone with significant homology to a Homo sapiens myosin II gene previously described.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: A study of the cell biology of motility in Eimeria tenella sporozoites
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Thesis digitised by ProQuest.
Keywords: Biological sciences; Eimeria tenella
URI: https://discovery.ucl.ac.uk/id/eprint/10098395
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