Probing Plasmodium falciparum sexual commitment at the single-cell level

Background: Malaria parasites go through major transitions during their complex life cycle, yet the underlying differentiation pathways remain obscure. Here we apply single cell transcriptomics to unravel the program inducing sexual differentiation in Plasmodium falciparum. Parasites have to make this essential life-cycle decision in preparation for human-to-mosquito transmission. Methods: By combining transcriptional profiling with quantitative imaging and genetics, we defined a transcriptional signature in sexually committed cells. Results: We found this transcriptional signature to be distinct from general changes in parasite metabolism that can be observed in response to commitment-inducing conditions. Conclusions: This proof-of-concept study provides a template to capture transcriptional diversity in parasite populations containing complex mixtures of different life-cycle stages and developmental programs, with important implications for our understanding of parasite biology and the ongoing malaria elimination campaign

The developmental cell biology of Trypanosoma brucei

Trypanosoma brucei provides an excellent system for studies of many aspects of cell biology, including cell structure and morphology, organelle positioning, cell division and protein trafficking. However, the trypanosome has a complex life cycle in which it must adapt either to the mammalian bloodstream or to different compartments within the tsetse fly. These differentiation events require stage-specific changes to basic cell biological processes and reflect responses to environmental stimuli and programmed differentiation events that must occur within a single cell. The organization of cell structure is fundamental to the trypanosome throughout its life cycle. Modulations of the overall cell morphology and positioning of the specialized mitochondrial genome, flagellum and associated basal body provide the classical descriptions of the different life cycle stages of the parasite. The dependency relationships that govern these morphological changes are now beginning to be understood and their molecular basis identified. The overall picture emerging is of a highly organized cell in which the rules established for cell division and morphogenesis in organisms such as yeast and mammalian cells do not necessarily apply. Therefore, understanding the developmental cell biology of the African trypanosome is providing insight into both fundamentally conserved and fundamentally different aspects of the organization of the eukaryotic cell

Essentialist Reasoning and Knowledge Effects on Biological Reasoning in Young Children

Biological kinds undergo a variety of changes during their life span, and these changes vary in degree by organism. Understanding that an organism, such as a caterpillar, maintains category identity over its life span despite dramatic changes is a key concept in biological reasoning. At present, we know little about the developmental trajectory of children’s understanding of dramatic life-cycle changes and how this might relate to their understanding of evolution. We suggest that this understanding is a key precursor to later understanding of evolutionary change. Two studies examined the impact of age and knowledge on children’s biological reasoning about living kinds that undergo a range of natural life-span changes—from subtle to dramatic. The participants, who were 3, 4, and 7 years old, were shown paired pictures of juvenile and adult animals and asked to endorse biological or nonbiological causal mechanisms to account for life-span change. Additionally, reasoning of 3- and 4-year-old participants was compared before and after exposure to caterpillars transforming into butterflies. The results are framed in terms of a developmental trajectory in essentialist reasoning, a cognitive bias that has been associated with difficulties in understanding and accepting evolution

Discovery of essential kinetoplastid-insect adhesion proteins and their function in Leishmania -sand fly interactions

Leishmania species, members of the kinetoplastid parasites, cause leishmaniasis, a neglected tropical disease, in millions of people worldwide. Leishmania has a complex life cycle with multiple developmental forms, as it cycles between a sand fly vector and a mammalian host; understanding their life cycle is critical to understanding disease spread. One of the key life cycle stages is the haptomonad form, which attaches to insect tissues through its flagellum. This adhesion, conserved across kinetoplastid parasites, is implicated in having an important function within their life cycles and hence in disease transmission. Here, we discover the kinetoplastid-insect adhesion proteins (KIAPs), which localise in the attached Leishmania flagellum. Deletion of these KIAPs impairs cell adhesion in vitro and prevents Leishmania from colonising the stomodeal valve in the sand fly, without affecting cell growth. Additionally, loss of parasite adhesion in the sand fly results in reduced physiological changes to the fly, with no observable damage of the stomodeal valve and reduced midgut swelling. These results provide important insights into a comprehensive understanding of the Leishmania life cycle, which will be critical for developing transmission-blocking strategies

Comparative embryology without a microscope: using genomic approaches to understand the evolution of development

Until recently, understanding developmental conservation and change has relied on embryological comparisons and analyses of single genes. Several studies, including one recently published in BMC Biology, have now taken a genomic approach to this classical problem, providing insights into how selection operates differentially across the life cycle

Proteomic differences between developmental stages of Toxoplasma gondii revealed by iTRAQ-based quantitative proteomics

Toxoplasma gondii has a complex two-host life-cycle between intermediate host and definitive host. Understanding proteomic variations across the life-cycle stages of T. gondii may improve the understanding of molecular adaption mechanism of T. gondii across life-cycle stages, and should have implications for the development of new treatment and prevention interventions against T. gondii infection. Here, we utilized LC–MS/MS coupled with iTRAQ labeling technology to identify differentially expressed proteins (DEPs) specific to tachyzoite (T), bradyzoites-containing cyst (C) and sporulated oocyst (O) stages of the cyst-forming T. gondii Prugniuad (Pru) strain. A total of 6285 proteins were identified in the three developmental stages of T. gondii. Our analysis also revealed 875, 656, and 538 DEPs in O vs. T, T vs. C, and C vs. O, respectively. The up- and down-regulated proteins were analyzed by Gene Ontology enrichment, KEGG pathway and STRING analyses. Some virulence-related factors and ribosomal proteins exhibited distinct expression patterns across the life-cycle stages. The virulence factors expressed in sporulated oocysts and the number of up-regulated virulence factors in the cyst stage were about twice as many as in tachyzoites. Of the 79 ribosomal proteins identified in T. gondii, the number of up-regulated ribosomal proteins was 33 and 46 in sporulated oocysts and cysts, respectively, compared with tachyzoites. These results support the hypothesis that oocyst and cystic stages are able to adapt to adverse environmental conditions and selection pressures induced by the host’s immune response, respectively. These findings have important implications for understanding of the developmental biology of T. gondii, which may facilitate the discovery of novel therapeutic targets to better control toxoplasmosis

Identification of transcription factors coupling the cell cycle machinery with environmental and developmental signals

In the recent years many studies contributed to our understanding of the mechanisms that drives the cell cycle machinery. Studies performed in yeast, animals, worms, flies and plants revealed that despite the evolutionary distance between these species, an universal picture can be drawn on how the basic cell cycle machinery is regulated. However, in spite of their highly conserved cell cycle machinery, it is remarkable how plants and animals have integrated the control of cell cycle differently into their specific developmental programs. In contrast to animals, plants develop mostly post-embryonic, which is characterized by continuous growth and organ formation during their entire life-span. This developmental style relies on the existence of stem cell niches within the root and shoot apical meristems, continuously supplying new cells. Next to this, plants are able to form organs de novo, like lateral roots, requiring cell cycle reactivation within already differentiated cells. Additionally, besides cell proliferation, endoreduplication plays as well an important role during different developmental processes. The plant body shows an amazing flexibility rendering them the ability to cope with different environmental and developmental signals. However, although different reports describe the influence of different environmental and developmental cues on cell cycle progression and endoreduplication, we have currently only limited knowledge on how these signals connect to the core cell cycle machinery. The aim of this project was to gain insight in how these intrinsic and extrinsic signals are integrated with the regulation of the cell cycle machinery. Although different core cell cycle genes display developmental and cell cycle-phase dependent transcriptional regulation, it is intriguing how little is known on their transcriptional regulators. Therefore the work was focused on determining new transcriptional regulators of core cell cycle genes and to try to connect these with specific environmental or developmental processes

Identification and stage-specific association with the translational apparatus of TbZFP3, a CCCH protein that promotes trypanosome life-cycle development

The post-transcriptional control of gene expression is becoming increasingly important in the understanding of regulated events in eukaryotic cells. The parasitic kinetoplastids have a unique reliance on such processes, because their genome is organized into polycistronic transcription units in which adjacent genes are not coordinately regulated. Indeed, the number of RNA-binding proteins predicted to be encoded in the genome of kinetoplastids is unusually large, invoking the presence of unique RNA regulators dedicated to gene expression in these evolutionarily ancient organisms. Here, we report that a small CCCH zinc finger protein, TbZFP3, enhances development between life-cycle stages in Trypanosoma brucei. Moreover, we demonstrate that this protein interacts both with the translational machinery and with other small CCCH proteins previously implicated in trypanosome developmental control. Antibodies to this protein also co-immunoprecipitate EP procyclin mRNA and encode the major surface antigen of insect forms of T. brucei. Strikingly, although TbZFP3 is constitutively expressed, it exhibits developmentally regulated association with polyribosomes, and mutational analysis demonstrates that this association is essential for the expression of phenotype. TbZFP3 is therefore a novel regulator of developmental events in kinetoplastids that acts at the level of the post-transcriptional control of gene expression

Systems biology approach to model the life cycle of Trypanosoma cruzi

Due to recent advances in reprogramming cell phenotypes, many efforts have been dedicated to developing reverse engineering procedures for the identification of gene regulatory networks that emulate dynamical properties associated with the cell fates of a given biological system. In this work, we propose a systems biology approach for the reconstruction of the gene regulatory network underlying the dynamics of the Trypanosoma cruzi's life cycle. By means of an optimisation procedure, we embedded the steady state maintenance, and the known phenotypic transitions between these steady states in response to environmental cues, into the dynamics of a gene network model. In the resulting network architecture we identified a small subnetwork, formed by seven interconnected nodes, that controls the parasite's life cycle. The present approach could be useful for better understanding other single cell organisms with multiple developmental stages.Revisión disponible en http://sedici.unlp.edu.ar/handle/10915/87345Centro Regional de Estudios Genómico

Applying family life cycle concepts in psychological practice with children and young people

Family life cycle models offer a framework for understanding family development and defining ‘normative’ transitions within this developmental progression. As such, family life cycle models have the potential to inform clinical practice: by allowing case presentations to be contextualised in terms of broader systemic and temporal processes, and possible disruptions to expected transitions. This article critically considers how family life cycle models, and the notion of normative transition, might be used to support psychological understanding and intervention when working clinically with children and young people. Models of normative development and transition may be helpfully integrated with broader psychological models to inform idiographic case formulation, and thereby influence practice. Although normative models can be used indicatively, care should be taken not to use these models prescriptively