A novel cytoplasmic structure containing DNA networks in Entamoeba histolytica trophozoites (original) (raw)
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Parasitology Research, 1988
We carried out a comparative study of nuclear and trophozoite diameters and of DNA thermal denaturation in eight Entamoeba strains cultured axenically (four of them E. histolytica, two initially designated as E. invadens, one E. moshkovskii, and one E. histolytica-like), as well as an analysis of the overall DNA sequence homology of the non-E. histolytica strains. The average nuclear (N) and trophozoite (T) diameters (in Μm) were, respectively: global averages ±SD for E. histolytica strains, 6.5±2.5 and 28.8±3.7; E. invadens IP101, 5.8 and 27.5, and PZ, 7.8 and 33.6; E. moshkovskii FIC, 4.1 and 12.9; E. histolytica- like Laredo strain, 5.0 and 20.6. The GC content of DNA, estimated by thermal elution in hydroxyapatite, was around 23% in HK9 and its clone HK9-1 and around 27% in the HM2 and HM3 E. histolytica strains; it was 37% in the Laredo strain, 26% in IP101, 35% in PZ, and 33% in FIC. The reassociation kinetics of PZ strain DNA showed that it consists of 40% repeated sequences and 60% unique sequences. By means of DNA association experiments in which one of each pair of DNAs tested had been labeled in vitro with 125I, we found the following overall sequence homology among the strains tested: PZ-FIC, 38%; IP101-Laredo, 38%; IP101-FIC, 47%; PZ-Laredo, 49%; Laredo-FIC, 69%; and IP101-PZ, 83%. We conclude that trophozoites of different E. histolytica strains have similar nuclear size and GC content, whereas these parameters and the nucleotide sequences are clearly different in every other Entamoeba species. Our data also suggest that PZ and IP101 strains do not belong to the same species.
Experimental Parasitology, 1978
We carried out a comparative study of nuclear and trophozoite diameters and of DNA thermal denaturation in eight Entamoeba strains cultured axenically (four of them E. histolytica, two initially designated as E. invadens, one E. moshkovskii, and one E. histolytica-like), as well as an analysis of the overall DNA sequence homology of the non-E, histolytica strains. The average nuclear (N) and trophozoite (T) diameters (in pro) were, respectively: global averages • SD for E. histolytica strains, 6.5 _+ 2.5 and 28.8 _+ 3.7 ; E. invadens IP101, 5.8 and 27.5, and PZ, 7.8 and 33.6; E. moshkovskii FIC, 4.1 and 12.9; E. histolyticalike Laredo strain, 5.0 and 20.6. The GC content of DNA, estimated by thermal elution in hydroxyapatite, was around 23% in HK9 and its clone HK9-1 and around 27% in the HM2 and HM3 E. histolytica strains; it was 37% in the Laredo strain, 26% in IPI01, 35% in PZ, and 33% in FIC. The reassociation kinetics of PZ strain DNA showed that it consists of 40% repeated sequences and 60% unique sequences. By means of DNA association experiments in which one of each pair of DNAs tested had been labeled in vitro with 125i, we found the following overall sequence homology among the strains tested: PZ-FIC, 38%; IPI01-Laredo, 38% ; IP101-FIC, 47% ; PZ-Laredo, 49% ; Laredo-FIC, 69%; and IPI01-PZ, 83%. We conclude that trophozoites of different E. histolytica strains have similar nuclear size and GC content, whereas these parameters and the nucleotide sequences are clearly different in every other Entamoeba species. Our data also suggest that PZ and IPl01 strains do not belong to the same species.
Molecular karyotype of Entamoeba histolytica and Entamoeba invadens
Transactions of the Royal Society of Tropical Medicine and Hygiene, 1990
We report the size-fractionation of Entamoeba histolvtica and E. invadens deoxvribonucleic acid (DNA) by pulsed field gradient electrophoresis. Using 3 different electrophoretic conditions, we were able to resolve 6 to 9 bands between 300 and 2000 kilobases (kb), distributed in 16-22 large and up to 31 small chromosomes for E. histolytica DNA. For E. invadens, 4 to 5 bands between 300 and over 2000 kb were resolved and discriminated in 4 large and 6 small chromosomes. A ribosomal probe from Trypanosoma brucei hybridized with a 1100 kb band in E. histolytica strain HMl:IMSS and with a 1000 kb band in clone A of strain HMl:IMSS. Both cell lines also showed hybridization at the origin. The ribosomal probe hybridized only at the origin of the E. invadens DNA lane. A triosephosphate isomerase DNA probe from T. brucei hybridized only with a 2000 kb band in E. invadens, indicating that banding patterns are chromosomal bands and ruling out the possibility of DNA degradation.
Entamoeba histolytica: DNA carrier vesicles in nuclei and kinetoplast-like organelles (EhkOs)
Molecular Genetics and Genomics, 2002
Entamoeba histolytica, the protozoan responsible for human amoebiasis, has a complex genome, whose linear chromosomes and DNA circles have so far eluded detailed analysis. We report the detection by transmission electron microscopy of nuclear vesicles (0.05-0.3 lm in diameter) carrying DNA in E. histolytica trophozoites. In late anaphase many of these nuclear vesicles were found to be organized in structures of 2.5·1 lm, in association with chromosomes and microtubules. In glutaraldehyde-fixed and detergenttreated trophozoites, nuclear vesicles displayed a nonmembranous envelope. Binding of phosphotungstate stain and recognition by serum from patients with systemic lupus erythematosus indicated that these vesicles contain DNA. Similar DNA carrier vesicles were found in the cytoplasm and in the E. histolytica kinetoplast-like organelle (EhkO). By Feulgen staining, we detected DNA carrier vesicles entering or leaving the nuclei, suggesting a structural relationship between the nuclear vesicles and the vesicles present in the EhkOs.
Microbiology-sgm, 2004
The existence of mitochondrion-related relict organelles (mitosomes) in the amitochondrial human pathogen Entamoeba histolytica and the detection of extranuclear DNA-containing cytoplasmic structures (EhKOs) has led to the suggestion that a remnant genome from the original mitochondrial endosymbiont might have been retained in this organism. This study reports on the mutually exclusive distribution of Cpn60 and extranuclear DNA in E. histolytica and on the distribution of Cpn60-containing mitosomes in this parasite. In situ nick-translation coupled to immunofluorescence microscopy failed to detect the presence of DNA in mitosomes, either in fixed parasite trophozoites or in partially purified organellar fractions. These results indicate that a remnant organellar genome has not been retained in E. histolytica mitosomes and demonstrate unequivocally that EhKOs and mitosomes are distinct and unrelated cellular structures.
Molecular and General Genetics, 1993
A highly variable DNA region (EhVRI), isolated from Entamoeba histolytica clone A, strain HM1:IMSS, is transcribed into several transcripts, which differ in genetically related clones. EhVRI (3.5 kb) is composed of two contiguous fragments; one of these 1.9 kb long, at the 3′ end, identified similar transcripts in clones A, L6 and C2 (all derived from strain HM 1: IMMS), the other of 1.6 kb, at the 5′ end, detected 0.5, 0.6 and 0.7 kb transcripts only in clone A. Variability of the 1.6 kb fragment was found even within the same clone maintained under different conditions. EhVRl was localized to 1.3 and 1.4 Mb linear chromosomes and also found in circular molecules. The sequence of the 1.6 kb fragment revealed the presence of a large number of different repeats, including inverted and palindromic repeats. A p145 sequence, previously detected in episomal DNA of the amoeba, was found at the 5′ end of EhVR1. The presence of EhVRI in linear and circular molecules, its high number of repeats, and its variability in genetically related clones suggest the existence of DNA regions that undergo dynamic non-reciprocal recombination between circular episomes and linear chromosomes, and may thus contribute to variability in the trophozoite genome.
DNA hybridization probe for clinical diagnosis of Entamoeba histolytica
Journal of Clinical Microbiology
As an alternative to microscopic identification of Entamoeba histolytica parasites isolated from -stool, a sensitive and species-specific DNA hybridization probe was made for rapid diagnosis of E. histolytica parasites in clinical samples directly applied to nylon membranes. The DNA hybridization probe was made by screening a genomic library of a virulent HM-1:IMSS strain of E. histolytica to detect recombinant plasmids containing highly repeated parasite DNA sequences. Four plasmid clones that reacted across Entamoeba species coded for highly repeated rRNA genes of E. histolytica. Four other plasmid clones were E. histolytica specific in that they bound to four axenized and nine xenic strains of E. histolytica but did not recognize closely related E. histolytica-like Laredo, Entamoeba moshkovskii, or Entamoeba invadens parasites. The diagnostic clones detected as few as eight cultured amoebae and did not distinguish between pathogenic and nonpathogenic zymodemes of E. histolytica. The diagnostic clones were sequenced and contained 145-base-pair sequences which appear to be tandemly repeated in the genome. No stable transcript which is homologous to the diagnostic DNA was detected. In a study of stool samples from Mexico City shown by microscopy to contain E. histolytica, Entamoeba coli, Giardia lamblia, Endolimax nana, Trichuris trichiuria, and Chilomastix mesnili parasites, the DNA hybridization probe demonstrated a sensitivity of 1.0 and a specificity of 0.93. We conclude that the DNA hybridization probe can be used for rapid and accurate diagnosis of E. histolytica parasites.
Entamoeba histolytica: ultrastructure of trophozoites recovered from experimental liver lesions
Experimental Parasitology, 2004
Ultrastructural studies on Entamoeba histolytica have been carried out mostly with trophozoites cultured for many years. Under these conditions, the availability of nutrients and the absence of environmental stimuli may switch off some phenotypic characteristics of the parasite. As a result, virulence of E. histolytica diminishes with prolonged culture passages, and the ability to form cysts disappears in axenically maintained