Influence of Micropatterned Grill Lines on Entamoeba histolytica Trophozoites Morphology and Migration

Abstract

Entamoeba histolytica, the causal agent of human amoebiasis, has two morphologically different phases: a resistant cyst and a trophozoite responsible for the invasion of the host tissues such as the colonic mucosa and the intestinal epithelium. During in vitro migration, trophozoites usually produce protuberances such as pseudopods and rarely filopodia, structures that have been observed in the interaction of trophozoites with human colonic epithelial tissue. To study the different membrane projections produced by the trophozoites, including pseudopods, filopodia, uropods, blebs, and others, we designed an induction system using erythrocyte extract or fibronectin (FN) in micropatterned grill lines (each micro-line containing multiple micro-portions of FN or erythrocyte extract) on which the trophozoites were placed in culture for migration assays. Using light, confocal, and scanning electron microscopy, we established that E. histolytica trophozoites frequently produce short and long filopodia, large retractile uropods in the rear, pseudopods, blebs, and others structures, also showing continuous migration periods. The present study provides a simple migration method to induce trophozoites to generate abundant membrane protrusion structures that are rarely obtained in normal or induced cultures, such as long filopodia; this method will allow a–better understanding of the interactions of trophozoites with FN and cell debris. E. histolytica trophozoites motility plays an important role in invasive amoebiasis. It has been proposed that both physical forces and chemical signals are involved in the trophozoite motility and migration. However, the in vivo molecules that drive the chemotactic migration remain to be determined. We propose the present assay to study host molecules that guide chemotactic behavior because the method is highly reproducible, and a live image of cell movement and migration can be quantified.

Lysine Methyltransferase EhPKMT2 Is Involved in the In Vitro Virulence of Entamoeba histolytica

“…Cell migration is positively or negatively regulated by different PKMTs; knockdown of SMYD3 provokes loss of cell migration [ 49 ], whereas knockout of SET7/9 improves it [ 37 ]. We evaluated the effect of EhPKMT2 knockdown on migration of E. histolytica trophozoites because this process is required to invade different tissues after disruption of the intestinal epithelium [ 50 ]. Interestingly, diminishing of EhPKMT2 expression induced an increase in cell migration.…”

Phytochemical Analysis and Amoebicidal Evaluation of Different Agave Species

“…In addition, it has been demonstrated that the motility of E. histolytica depends on the sustained instability of the intracellular hydrostatic pressure that drives the cyclic generation and healing of membrane blebs [69]. Furthermore, actin filaments constitute the physical backbone of membrane protrusions [70]. The prevalence of the filopodial protrusions that A. maximiliana caused in trophozoites could be related to alterations in intracellular hydrostatic pressure and the actin cytoskeleton.…”

Analisis Faktor Resiko Kontaminasi Entamoeba Histolytica pada Air Sumur Masayarakat Desa Ujong Blang Kecamatan Banda Sakti Kota Lhokseumawe

“…Hasil laboratorium pada penelitian ini menunjukkan bahwa tidak ditemukan adanya kista Entamoeba histolytica pada seluruh sampel air sumur meskipun suhu air sumur tidak ekstrim (≤ 32˚C) dan pH air sumur ≤ 8. Pada pemeriksaan mikroskopik menggunakan cairan lugol tidak ditemukan adanya kemiripan secara morfologi dan morfometri dengan kista Entamoeba histolytica akan tetapi didapatkan adanya telur Hookworm, larva Rabdiform nematoda dan ciliata pada beberapa sampel air sumur (10).…”