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An Insight into the proteome of Crithidia fasciculata choanomastigotes as a comparative approach to axenic growth, peanut lectin agglutination and differentiation of Leishmania spp. promastigotes.

Alcolea PJ, Alonso A, García-Tabares F, Toraño A, Larraga V - PLoS ONE (2014)

Bottom Line: A ground-breaking analysis of differential protein abundance in Crithidia fasciculata is reported herein.The comparison of the outcome with previous gene expression profiling studies developed in the related human pathogens of the genus Leishmania has revealed substantial differences between the motile stages of these closely related organisms in abundance of proteins involved in catabolism, redox homeostasis, intracellular signalling, and gene expression regulation.The result is that choanomastigotes are able to agglutinate with peanut lectin and a non-agglutinating subpopulation can be also isolated.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Microbiology and Biology of Infections and Service of Proteomics and Genomics, Centro de Investigaciones Biológicas (Consejo Superior de Investigaciones Científicas), Madrid, Spain.

ABSTRACT
The life cycle of the trypanosomatid Crithidia fasciculata is monogenetic, as the unique hosts of these parasites are different species of culicids. The comparison of these non-pathogenic microorganisms evolutionary close to other species of trypanosomatids that develop digenetic life cycles and cause chronic severe sickness to millions of people worldwide is of outstanding interest. A ground-breaking analysis of differential protein abundance in Crithidia fasciculata is reported herein. The comparison of the outcome with previous gene expression profiling studies developed in the related human pathogens of the genus Leishmania has revealed substantial differences between the motile stages of these closely related organisms in abundance of proteins involved in catabolism, redox homeostasis, intracellular signalling, and gene expression regulation. As L. major and L. infantum agglutinate with peanut lectin and non-agglutinating parasites are more infective, the agglutination properties were evaluated in C. fasciculata. The result is that choanomastigotes are able to agglutinate with peanut lectin and a non-agglutinating subpopulation can be also isolated. As a difference with L. infantum, the non-agglutinating subpopulation over-expresses the whole machinery for maintenance of redox homeostasis and the translation factors eIF5a, EF1α and EF2, what suggests a relationship between the lack of agglutination and a differentiation process.

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Life cycle, growth kinetics and peanut lectin agglutination of C. fasciculata choanomastigotes.(A) The monogenetic life cycle of C. fasciculata involves a culicid host, where amastigotes attached to the gut epithelium and voided in faeces are disseminated in the environment and orally passed to other hosts at any of the developmental stages. Choanomastigotes are the motile stage that allows the colonization of the gut of the host. GE: gut epithelium. Adapted from Olsen, 1974. (B) Average growth curve of three C. fasciculata choanomastigote cultures (three biological replicates). Total proteins were extracted every day until the culture reached the stationary phase. N is the average cell density. (C) and (D) 10% Giemsa staining of the PNA+ and PNA-C. fasciculata choanomastigote subpopulations within the stationary phase of axenic culture, respectively.
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pone-0113837-g001: Life cycle, growth kinetics and peanut lectin agglutination of C. fasciculata choanomastigotes.(A) The monogenetic life cycle of C. fasciculata involves a culicid host, where amastigotes attached to the gut epithelium and voided in faeces are disseminated in the environment and orally passed to other hosts at any of the developmental stages. Choanomastigotes are the motile stage that allows the colonization of the gut of the host. GE: gut epithelium. Adapted from Olsen, 1974. (B) Average growth curve of three C. fasciculata choanomastigote cultures (three biological replicates). Total proteins were extracted every day until the culture reached the stationary phase. N is the average cell density. (C) and (D) 10% Giemsa staining of the PNA+ and PNA-C. fasciculata choanomastigote subpopulations within the stationary phase of axenic culture, respectively.

Mentions: Although these parasites are polymorphic, two stages are clearly distinguished. Choanomastigotes are free-swimming stumpy cells characteristic of this genus that are round in their posterior part and truncated in the apical pole by the funnel-shaped flagellar pocket close to the kinetoplast, which is slightly anterior to the nucleus. Amastigotes are non-motile round cells with a flagellum non-emergent from the cellular body. Therefore, they are morphologically similar to amastigotes of the genus Leishmania, although they are extracellular (reviewed in [2]). The life cycle of C. fasciculata is developed in the gut of the culicid, which becomes infected by ingestion of amastigotes voided with feces of other hosts. Then, amastigotes undergo a differentiation process into choanomastigotes to ensure proper colonization of the gut. Choanomastigotes differentiate back into non-motile round amastigotes that are attached to the gut epithelium by hemidesmosomes [3] frequently leading to damage [4]. Infected adult mosquitoes contaminate aquatic environments with amastigotes as well as flowers when they feed on nectar, thus providing chances for transmission of the parasite. Amastigotes are released within the feces or the entire body of the dead insect. Eventually, the larval and pupal instars of mosquitoes get infected in the aquatic habitat and finally amastigotes are transmitted to the adult mosquito through the metamorphosing gut [2] leading to completion of the life cycle (Fig. 1A).


An Insight into the proteome of Crithidia fasciculata choanomastigotes as a comparative approach to axenic growth, peanut lectin agglutination and differentiation of Leishmania spp. promastigotes.

Alcolea PJ, Alonso A, García-Tabares F, Toraño A, Larraga V - PLoS ONE (2014)

Life cycle, growth kinetics and peanut lectin agglutination of C. fasciculata choanomastigotes.(A) The monogenetic life cycle of C. fasciculata involves a culicid host, where amastigotes attached to the gut epithelium and voided in faeces are disseminated in the environment and orally passed to other hosts at any of the developmental stages. Choanomastigotes are the motile stage that allows the colonization of the gut of the host. GE: gut epithelium. Adapted from Olsen, 1974. (B) Average growth curve of three C. fasciculata choanomastigote cultures (three biological replicates). Total proteins were extracted every day until the culture reached the stationary phase. N is the average cell density. (C) and (D) 10% Giemsa staining of the PNA+ and PNA-C. fasciculata choanomastigote subpopulations within the stationary phase of axenic culture, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263474&req=5

pone-0113837-g001: Life cycle, growth kinetics and peanut lectin agglutination of C. fasciculata choanomastigotes.(A) The monogenetic life cycle of C. fasciculata involves a culicid host, where amastigotes attached to the gut epithelium and voided in faeces are disseminated in the environment and orally passed to other hosts at any of the developmental stages. Choanomastigotes are the motile stage that allows the colonization of the gut of the host. GE: gut epithelium. Adapted from Olsen, 1974. (B) Average growth curve of three C. fasciculata choanomastigote cultures (three biological replicates). Total proteins were extracted every day until the culture reached the stationary phase. N is the average cell density. (C) and (D) 10% Giemsa staining of the PNA+ and PNA-C. fasciculata choanomastigote subpopulations within the stationary phase of axenic culture, respectively.
Mentions: Although these parasites are polymorphic, two stages are clearly distinguished. Choanomastigotes are free-swimming stumpy cells characteristic of this genus that are round in their posterior part and truncated in the apical pole by the funnel-shaped flagellar pocket close to the kinetoplast, which is slightly anterior to the nucleus. Amastigotes are non-motile round cells with a flagellum non-emergent from the cellular body. Therefore, they are morphologically similar to amastigotes of the genus Leishmania, although they are extracellular (reviewed in [2]). The life cycle of C. fasciculata is developed in the gut of the culicid, which becomes infected by ingestion of amastigotes voided with feces of other hosts. Then, amastigotes undergo a differentiation process into choanomastigotes to ensure proper colonization of the gut. Choanomastigotes differentiate back into non-motile round amastigotes that are attached to the gut epithelium by hemidesmosomes [3] frequently leading to damage [4]. Infected adult mosquitoes contaminate aquatic environments with amastigotes as well as flowers when they feed on nectar, thus providing chances for transmission of the parasite. Amastigotes are released within the feces or the entire body of the dead insect. Eventually, the larval and pupal instars of mosquitoes get infected in the aquatic habitat and finally amastigotes are transmitted to the adult mosquito through the metamorphosing gut [2] leading to completion of the life cycle (Fig. 1A).

Bottom Line: A ground-breaking analysis of differential protein abundance in Crithidia fasciculata is reported herein.The comparison of the outcome with previous gene expression profiling studies developed in the related human pathogens of the genus Leishmania has revealed substantial differences between the motile stages of these closely related organisms in abundance of proteins involved in catabolism, redox homeostasis, intracellular signalling, and gene expression regulation.The result is that choanomastigotes are able to agglutinate with peanut lectin and a non-agglutinating subpopulation can be also isolated.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Microbiology and Biology of Infections and Service of Proteomics and Genomics, Centro de Investigaciones Biológicas (Consejo Superior de Investigaciones Científicas), Madrid, Spain.

ABSTRACT
The life cycle of the trypanosomatid Crithidia fasciculata is monogenetic, as the unique hosts of these parasites are different species of culicids. The comparison of these non-pathogenic microorganisms evolutionary close to other species of trypanosomatids that develop digenetic life cycles and cause chronic severe sickness to millions of people worldwide is of outstanding interest. A ground-breaking analysis of differential protein abundance in Crithidia fasciculata is reported herein. The comparison of the outcome with previous gene expression profiling studies developed in the related human pathogens of the genus Leishmania has revealed substantial differences between the motile stages of these closely related organisms in abundance of proteins involved in catabolism, redox homeostasis, intracellular signalling, and gene expression regulation. As L. major and L. infantum agglutinate with peanut lectin and non-agglutinating parasites are more infective, the agglutination properties were evaluated in C. fasciculata. The result is that choanomastigotes are able to agglutinate with peanut lectin and a non-agglutinating subpopulation can be also isolated. As a difference with L. infantum, the non-agglutinating subpopulation over-expresses the whole machinery for maintenance of redox homeostasis and the translation factors eIF5a, EF1α and EF2, what suggests a relationship between the lack of agglutination and a differentiation process.

Show MeSH
Related in: MedlinePlus