Giardia flagellar motility is not directly required to maintain attachment to surfaces

Autor: Jonathan K Pham, David J. Richter, Scott C. Dawson, Susan A. House
Přispěvatelé: Sibley, L David
Jazyk: angličtina
Rok vydání: 2011
Předmět:
Zdroj: PLoS Pathogens, Vol 7, Iss 8, p e1002167 (2011)
PLoS Pathogens
PLoS pathogens, vol 7, iss 8
ISSN: 1553-7374
1553-7366
Popis: Giardia trophozoites attach to the intestinal microvilli (or inert surfaces) using an undefined “suction-based” mechanism, and remain attached during cell division to avoid peristalsis. Flagellar motility is a key factor in Giardia's pathogenesis and colonization of the host small intestine. Specifically, the beating of the ventral flagella, one of four pairs of motile flagella, has been proposed to generate a hydrodynamic force that results in suction-based attachment via the adjacent ventral disc. We aimed to test this prevailing “hydrodynamic model” of attachment mediated by flagellar motility. We defined four distinct stages of attachment by assessing surface contacts of the trophozoite with the substrate during attachment using TIRF microscopy (TIRFM). The lateral crest of the ventral disc forms a continuous perimeter seal with the substrate, a cytological indication that trophozoites are fully attached. Using trophozoites with two types of molecularly engineered defects in flagellar beating, we determined that neither ventral flagellar beating, nor any flagellar beating, is necessary for the maintenance of attachment. Following a morpholino-based knockdown of PF16, a central pair protein, both the beating and morphology of flagella were defective, but trophozoites could still initiate proper surface contacts as seen using TIRFM and could maintain attachment in several biophysical assays. Trophozoites with impaired motility were able to attach as well as motile cells. We also generated a strain with defects in the ventral flagellar waveform by overexpressing a dominant negative form of alpha2-annexin::GFP (D122A, D275A). This dominant negative alpha2-annexin strain could initiate attachment and had only a slight decrease in the ability to withstand normal and shear forces. The time needed for attachment did increase in trophozoites with overall defective flagellar beating, however. Thus while not directly required for attachment, flagellar motility is important for positioning and orienting trophozoites prior to attachment. Drugs affecting flagellar motility may result in lower levels of attachment by indirectly limiting the number of parasites that can position the ventral disc properly against a surface and against peristaltic flow.
Author Summary Giardia is a widespread, single-celled, intestinal parasite that infects millions of people and animals each year. Colonization of the small intestine is a critical part of Giardia's life cycle in any host. This colonization is initiated when cells attach to the intestinal wall via a specialized suction cup-like structure, the ventral disc. In the host, Giardia moves by beating four pairs of flagella; movement of the ventral pair has been implicated in attachment. This study shows that the beating of the flagella is not important for attachment, but rather for positioning Giardia close to the intestinal wall prior to attachment, and thus disproves the commonly held model of giardial attachment. This work implies that drugs targeting Giardia motility could prevent or slow attachment, leading to lower rates of infection.
Databáze: OpenAIRE
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