1989 ◽  
Vol 109 (5) ◽  
pp. 2323-2335 ◽  
Author(s):  
D A Peattie ◽  
R A Alonso ◽  
A Hein ◽  
J P Caulfield

The giardins are a group of 29-38-kD proteins in the ventral disk of the protozoan parasite Giardia lamblia. The disk attaches the parasite to the host's intestinal epithelium and is composed of parallel, coiled microtubules that are adjacent to the ventral plasma membrane and from which processes called microribbons extend into the cytoplasm; the microribbons are connected by crossbridges. G. lamblia cytoskeletons, consisting of disks and attached flagella, were isolated and used to show that the 29-38-kD proteins separate into five bands by one-dimensional electrophoresis and into 23 species by two-dimensional analysis. Rabbit antibodies raised against a 33-kD protein band, purified by one-dimensional gel electrophoresis and shown to contain three proteins by two-dimensional electrophoresis, recognized 17 proteins by two-dimensional immunoblot analysis. By immunofluorescence these antibodies reacted with the ventral disk but not with the flagella in isolated cytoskeletons. Electron microscopy revealed that the anti-giardin antibodies bound to the edges of the microribbons but not to the microtubules, crossbridges, or other, nondisk structures. Antibodies to tubulin reacted with both the disk and flagella in isolated cytoskeletons but bound only to the microtubules in these structures. The amino-terminal sequence of the 33-kD immunogen was determined and used to construct a DNA oligomer, and the oligomer was used to isolate the alpha giardin gene. The gene was used to hybrid select RNA, and the in vitro translation product from this RNA was precipitated by the antibodies against the 33-kD immunogen. The gene sequence was a single open reading frame of 885 nucleotides that predicted a protein of 33.8 kD. The protein sequence is unique, having no significant homology to two other giardin sequences or to any sequences within the Protein Identification Resource. It is predicted to be 82% alpha helical. The downstream sequence of the gene indicates that the sequence AGT-PuAA is located six to nine nucleotides beyond the stop codon in all protein-encoding genes of G. lamblia that have been sequenced and reported to date.


2020 ◽  
Vol 13 (12) ◽  
pp. 440
Author(s):  
Verónica Yadira Ochoa-Maganda ◽  
Itzia Azucena Rangel-Castañeda ◽  
Daniel Osmar Suárez-Rico ◽  
Rafael Cortés-Zárate ◽  
José Manuel Hernández-Hernández ◽  
...  

Giardia lamblia is a flagellated protozoan responsible for giardiasis, a worldwide diarrheal disease. The adverse effects of the pharmacological treatments and the appearance of drug resistance have increased the rate of therapeutic failures. In the search for alternative therapeutics, drug repositioning has become a popular strategy. Acetylsalicylic acid (ASA) exhibits diverse biological activities through multiple mechanisms. However, the full spectrum of its activities is incompletely understood. In this study we show that ASA displayed direct antigiardial activity and affected the adhesion and growth of trophozoites in a time-dose-dependent manner. Electron microscopy images revealed remarkable morphological alterations in the membrane, ventral disk, and caudal region. Using mass spectrometry and real-time quantitative reverse transcription (qRT-PCR), we identified that ASA induced the overexpression of heat shock protein 70 (HSP70). ASA also showed a significant increase of five ATP-binding cassette (ABC) transporters (giABC, giABCP, giMDRP, giMRPL and giMDRAP1). Additionally, we found low toxicity on Caco-2 cells. Taken together, these results suggest an important role of HSPs and ABC drug transporters in contributing to stress tolerance and protecting cells from ASA-induced stress.


1999 ◽  
Vol 85 (12) ◽  
pp. 951-955 ◽  
Author(s):  
Adriana Lanfredi-Rangel ◽  
José A. Diniz Jr ◽  
Wanderley de Souza
Keyword(s):  

1981 ◽  
Vol 47 (1) ◽  
pp. 139-166 ◽  
Author(s):  
D.V. Holberton ◽  
A.P. Ward

The sucking disk of Giardia is supported by a large, plate-like organelle: the ventral disk cytoskeleton. Extraction by Triton-X 100 of Giardia trophozoites from the mouse gut, or of G. duodenalis or G. lamblia grown from cultures, yields cell-free disk cytoskeletons. Up to 8 flagellar axonemes may be attached to an isolated disk. Disks are seen in the electron microscope to be composed of concentrically coiled microtubules bonded to microribbons. Microribbons are large, laminated structures, linked by dense networks of crossbridges. They are made up of regularly arranged subunits. Microtubules and microribbons are preserved in Triton for long periods, but crossbridges are slowly dissolved. Whereas the addition of ATP causes axonemes to resume bending, active movements were not detected in disks. It seems more likely that a disk is a passive effector, which may be acted upon by other contractile structures of the cytoplasm. It is highly specialized and quintessential cytoskeleton. Disks and axonemes will dissolve in sodium dodecyl sulphate (SDS), and after SDS-gel electrophoresis 2 prominent bands are apparent. One, corresponding to tubulin, migrates in low ionic strength, high pH buffers as 2 closely spaced bands of equal staining density. The other, a smaller protein, is a complex polypeptide band of molecular weight 30 000 Daltons. Allowing for staining differences, the 2 proteins are probably present in cytoskeletons in roughly equal amounts. Because of their size microribbons account for at least 75% of the structured material stained by electron stains in pellets of cytoskeletons. Disk and axoneme microtubules comprise the minority fraction. This result suggests that, like microtubules, microribbons are a source of structural tubulin and probably also contain the 30 000 mol. wt protein.


Zootaxa ◽  
2017 ◽  
Vol 4282 (2) ◽  
pp. 337 ◽  
Author(s):  
SOO JEONG LEE ◽  
JIN-KOO KIM ◽  
YOSHIAKI KAI ◽  
SHIN’ICHIRO IKEGUCHI ◽  
TETSUJI NAKABO

A taxonomic review of the dwarf species of the genus Eumicrotremus (Actinopterygii: Cottoidei: Cyclopteridae), previously recognized as belonging to Lethotremus, established the existence of three species, viz. Eumicrotremus awae (Jordan and Snyder, 1902), Eumicrotremus uenoi sp. nov., and Eumicrotremus jindoensis sp. nov., from the western North Pacific. Eumicrotremus awae, known from the Pacific coast of Honshu Is. (Chiba southward to Mie), is characterized by the absence of spiny tubercles or fleshy papillae on the body, the anterior three mandibular pores each with a barbel-like tube, interorbital and suborbital pores usually absent, the opercular flap rounded, many papillae present on the ventral disk and the caudal-fin relatively short, 21.0–30.5% of SL (standard length). Eumicrotremus uenoi, known from the western coast of Honshu Is. and the southern coast of Korean Peninsula, is characterized by the absence of spiny tubercles or fleshy papillae on the body (or weak if present), the anterior three mandibular pores each with a barbel-like tube, interorbital and suborbital pores usually present, the opercular flap rounded, the ventral disk with many papillae and the caudal-fin relatively short, 19.5–31.4% of SL. Eumicrotremus jindoensis, known from the southwestern coast of Korean Peninsula, and the coasts of Yellow Sea, is characterized by a smooth soft body, triangular opercular flap, absence of papillae on the ventral disk and a long caudal fin, 32.7–42.1% of SL. Sequence variations of the mictochondrial DNA cytochrome c oxidase subunit I (mtDNA COI) and the nuclear myh6 genes supported the validity of the three species. 


2020 ◽  
Vol 31 (15) ◽  
pp. 1611-1622
Author(s):  
Kelly M. Hennessey ◽  
Germain C. M. Alas ◽  
Ilse Rogiers ◽  
Renyu Li ◽  
Ethan A. Merritt ◽  
...  

Here we study the role of Nek8445 in regulating cell division and microtubule array organization in Giardia. Depletion of Nek8445 results in 87% of cells being stalled or blocked in cytokinesis. Nek8445 regulates ventral disk organization, funis formation, axoneme exit, and cell shape, all of which contribute to the observed cytokinesis defects.


BIOCELL ◽  
2003 ◽  
Vol 27 (3) ◽  
pp. 329-346
Author(s):  
ALBERTO J. SOLARI ◽  
MONICA I. RAHN ◽  
ALICIA SAURA ◽  
HUGO D. LUJAN

1973 ◽  
Vol 13 (1) ◽  
pp. 11-41 ◽  
Author(s):  
D. V. HOLBERTON

The topography of the Giardia trophozoite is dominated by the large domed sucking disk of the ventral surface. Attached to the host duodenal epithelium, the rim of this disk penetrates the enteric surface coat and interdigitates with microvilli of the epithelial cells, approaching to within 20 nm of the host surface membrane. Distortion of the host brush border within the disk suggests an applied suction force. A mechanical explanation of disk action is sought in a detailed description of the fine structure of components of the ventral surface - but is found to be untenable. The disk is supported by a platform of modified 25-nm microtubules, linked to the ventral membrane by side arms and bearing heavily cross-linked vertical dense ribbons. It is argued that such is the architecture of rigidity rather than relative movement. Around the disk a mobile cytoplasmic flange is supported by 2 lateral plates of periodic substructure. The flange has no clear mechanical role in attachment; a likely evolutionary origin from a component of the anterior axonemal axis is suggested. The cavity of the ventral disk leads posteriorly through a portal into the ventrocaudal groove: a shallow depression that houses the ventral flagella. Observation of isolated living trophozoites suggests that attachment depends on the continuing activity of the ventral flagella, which normally beat synchronously in a sinusoidal waveform. Electron micrographs confirm that this waveform is maintained in situ on the host epithelium. Of the 4 pairs of flagella, the ultrastructure of the ventral flagella is notable for additional components in the flagellar shaft, including an intraflagellar dense rod linked to 3 axonemal doublets by fine connectives. From a consideration of analogous macroscopic systems, a preliminary hydrodynamic analysis is advanced in which the suction force of attachment follows from the pattern of fluid flow induced by the beating ventral flagella. The significance of the conclusion that cytoplasmic microtubules (or structures derived from them) apparently maintain cell shape in the face of an applied external force is discussed.


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