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An atlas for Schistosoma mansoni organs and life-cycle stages using cell type-specific markers and confocal microscopy.

Collins JJ, King RS, Cogswell A, Williams DL, Newmark PA - PLoS Negl Trop Dis (2011)

Bottom Line: This analysis uncovered more than 20 new markers that label most cercarial tissues, including the tegument, the musculature, the protonephridia, the secretory system and the nervous system.Examining the effectiveness of a subset of these markers in S. mansoni adults and miracidia, we demonstrate the value of these tools for labeling tissues in a variety of life-cycle stages.The methodologies described here will facilitate functional analyses aimed at understanding fundamental biological processes in these parasites.

View Article: PubMed Central - PubMed

Affiliation: Howard Hughes Medical Institute, Department of Cell and Developmental Biology, Neuroscience Program, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.

ABSTRACT
Schistosomiasis (bilharzia) is a tropical disease caused by trematode parasites (Schistosoma) that affects hundreds of millions of people in the developing world. Currently only a single drug (praziquantel) is available to treat this disease, highlighting the importance of developing new techniques to study Schistosoma. While molecular advances, including RNA interference and the availability of complete genome sequences for two Schistosoma species, will help to revolutionize studies of these animals, an array of tools for visualizing the consequences of experimental perturbations on tissue integrity and development needs to be made widely available. To this end, we screened a battery of commercially available stains, antibodies and fluorescently labeled lectins, many of which have not been described previously for analyzing schistosomes, for their ability to label various cell and tissue types in the cercarial stage of S. mansoni. This analysis uncovered more than 20 new markers that label most cercarial tissues, including the tegument, the musculature, the protonephridia, the secretory system and the nervous system. Using these markers we present a high-resolution visual depiction of cercarial anatomy. Examining the effectiveness of a subset of these markers in S. mansoni adults and miracidia, we demonstrate the value of these tools for labeling tissues in a variety of life-cycle stages. The methodologies described here will facilitate functional analyses aimed at understanding fundamental biological processes in these parasites.

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The nervous system of the cercarial tail.(A) Extensive neural projections within the tail visualized by β-tubulin immunostaining. Overlay with phalloidin and DAPI show the position of the nerves relative to the tail musculature and nuclei, respectively. (B) Superficial neural projections (green) laying outside muscle layer (magenta). Arrowhead indicates a sensory papilla. (C) Longitudinal nerve cord (white arrowhead) running along a longitudinal muscle within the tail. Scale bars, 10 µm. Anterior faces left in all panels.
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pntd-0001009-g005: The nervous system of the cercarial tail.(A) Extensive neural projections within the tail visualized by β-tubulin immunostaining. Overlay with phalloidin and DAPI show the position of the nerves relative to the tail musculature and nuclei, respectively. (B) Superficial neural projections (green) laying outside muscle layer (magenta). Arrowhead indicates a sensory papilla. (C) Longitudinal nerve cord (white arrowhead) running along a longitudinal muscle within the tail. Scale bars, 10 µm. Anterior faces left in all panels.

Mentions: In addition to anti-synapsin staining, we found that anti-β-tubulin was useful for detecting the nervous system within the tail (Figure 5A, B, C). However, unlike anti-synapsin that labels synaptic regions, anti-β-tubulin stained microtubule-rich neural projections throughout the tail. These fine projections could be observed transversely projecting through the interior of the tail (Figure 5A), superficially between the tegument and the muscles (Figure 5B), and within a pair of cords beneath the longitudinal muscle fibers (Figure 5C).


An atlas for Schistosoma mansoni organs and life-cycle stages using cell type-specific markers and confocal microscopy.

Collins JJ, King RS, Cogswell A, Williams DL, Newmark PA - PLoS Negl Trop Dis (2011)

The nervous system of the cercarial tail.(A) Extensive neural projections within the tail visualized by β-tubulin immunostaining. Overlay with phalloidin and DAPI show the position of the nerves relative to the tail musculature and nuclei, respectively. (B) Superficial neural projections (green) laying outside muscle layer (magenta). Arrowhead indicates a sensory papilla. (C) Longitudinal nerve cord (white arrowhead) running along a longitudinal muscle within the tail. Scale bars, 10 µm. Anterior faces left in all panels.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3050934&req=5

pntd-0001009-g005: The nervous system of the cercarial tail.(A) Extensive neural projections within the tail visualized by β-tubulin immunostaining. Overlay with phalloidin and DAPI show the position of the nerves relative to the tail musculature and nuclei, respectively. (B) Superficial neural projections (green) laying outside muscle layer (magenta). Arrowhead indicates a sensory papilla. (C) Longitudinal nerve cord (white arrowhead) running along a longitudinal muscle within the tail. Scale bars, 10 µm. Anterior faces left in all panels.
Mentions: In addition to anti-synapsin staining, we found that anti-β-tubulin was useful for detecting the nervous system within the tail (Figure 5A, B, C). However, unlike anti-synapsin that labels synaptic regions, anti-β-tubulin stained microtubule-rich neural projections throughout the tail. These fine projections could be observed transversely projecting through the interior of the tail (Figure 5A), superficially between the tegument and the muscles (Figure 5B), and within a pair of cords beneath the longitudinal muscle fibers (Figure 5C).

Bottom Line: This analysis uncovered more than 20 new markers that label most cercarial tissues, including the tegument, the musculature, the protonephridia, the secretory system and the nervous system.Examining the effectiveness of a subset of these markers in S. mansoni adults and miracidia, we demonstrate the value of these tools for labeling tissues in a variety of life-cycle stages.The methodologies described here will facilitate functional analyses aimed at understanding fundamental biological processes in these parasites.

View Article: PubMed Central - PubMed

Affiliation: Howard Hughes Medical Institute, Department of Cell and Developmental Biology, Neuroscience Program, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.

ABSTRACT
Schistosomiasis (bilharzia) is a tropical disease caused by trematode parasites (Schistosoma) that affects hundreds of millions of people in the developing world. Currently only a single drug (praziquantel) is available to treat this disease, highlighting the importance of developing new techniques to study Schistosoma. While molecular advances, including RNA interference and the availability of complete genome sequences for two Schistosoma species, will help to revolutionize studies of these animals, an array of tools for visualizing the consequences of experimental perturbations on tissue integrity and development needs to be made widely available. To this end, we screened a battery of commercially available stains, antibodies and fluorescently labeled lectins, many of which have not been described previously for analyzing schistosomes, for their ability to label various cell and tissue types in the cercarial stage of S. mansoni. This analysis uncovered more than 20 new markers that label most cercarial tissues, including the tegument, the musculature, the protonephridia, the secretory system and the nervous system. Using these markers we present a high-resolution visual depiction of cercarial anatomy. Examining the effectiveness of a subset of these markers in S. mansoni adults and miracidia, we demonstrate the value of these tools for labeling tissues in a variety of life-cycle stages. The methodologies described here will facilitate functional analyses aimed at understanding fundamental biological processes in these parasites.

Show MeSH
Related in: MedlinePlus