Rapid determination of nematode cell and organ susceptibility to toxic treatments
In research focused on the intestine of parasitic nematodes, we recently identified small molecule inhibitors toxic to intestinal cells of larval Ascaris suum (nematode intestinal toxins/toxicants; “NITs”). Some NITs had anthelmintic activity across the phylogenetic diversity of the Nematoda. The wh...
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doaj-91e4c6a205704d8c82dd783b5524d2262020-12-17T04:48:09ZengElsevierInternational Journal for Parasitology: Drugs and Drug Resistance2211-32072020-12-0114167182Rapid determination of nematode cell and organ susceptibility to toxic treatmentsDouglas P. Jasmer0Bruce A. Rosa1Rahul Tyagi2Makedonka Mitreva3Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, 99164, USADivision of Infectious Diseases, Department of Medicine, Washington University School of Medicine, St. Louis, MO, 63110, USADivision of Infectious Diseases, Department of Medicine, Washington University School of Medicine, St. Louis, MO, 63110, USADivision of Infectious Diseases, Department of Medicine, Washington University School of Medicine, St. Louis, MO, 63110, USA; Department of Genetics, Washington University School of Medicine, St. Louis, MO 63110, St. Louis, MO, 63110, USA; McDonnell Genome Institute, Washington University School of Medicine, St. Louis, Missouri, 63108, USA; Corresponding author. Division of Infectious Diseases, Department of Medicine, Washington University School of Medicine, St. Louis, MO, 63110, USA.In research focused on the intestine of parasitic nematodes, we recently identified small molecule inhibitors toxic to intestinal cells of larval Ascaris suum (nematode intestinal toxins/toxicants; “NITs”). Some NITs had anthelmintic activity across the phylogenetic diversity of the Nematoda. The whole-worm motility inhibition assay quantified anthelmintic activity, but worm responses to NITs in relation to pathology or affected molecular pathways was not acquired. In this study we extended this research to more comprehensively determine in whole larval A. suum the cells, organ systems, molecular targets, and potential cellular pathways involved in mechanisms of toxicity leading to cell death. The experimental system utilized fluorescent nuclear probes (bisbenzimide, propidium iodide), NITs, an A. suum larval parasite culture system and transcriptional responses (RNA-seq) to NITs. The approach provides for rapid resolution of NIT-induced cell death among organ systems (e.g. intestine, excretory, esophagus, hypodermis and seam cells, and nervous), discriminates among NITs based on cell death profiles, and identifies cells and organ systems with the greatest NIT sensitivity (e.g. intestine and apparent neuronal cells adjacent to the nerve ring). Application was extended to identify cells and organs sensitive to several existing anthelmintics. This approach also resolved intestinal cell death and irreparable damage induced in adult A. suum by two NITs, establishing a new model to elucidate relevant pathologic mechanisms in adult worms. RNA-seq analysis resolved A. suum genes responsive to treatments with three NITs, identifying dihydroorotate dehydrogenase (uridine synthesis) and RAB GTPase(s) (vesicle transport) as potential targets/pathways leading to cell death. A set of genes induced by all three NITs tested suggest common stress or survival responses activated by NITs. Beyond the presented specific lines of research, elements of the overall experimental system presented in this study have broad application toward systematic development of new anthelmintics.http://www.sciencedirect.com/science/article/pii/S2211320720300373NematodeIntestineAnthelminticsMicroscopyPathologyRNA-Seq |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Douglas P. Jasmer Bruce A. Rosa Rahul Tyagi Makedonka Mitreva |
spellingShingle |
Douglas P. Jasmer Bruce A. Rosa Rahul Tyagi Makedonka Mitreva Rapid determination of nematode cell and organ susceptibility to toxic treatments International Journal for Parasitology: Drugs and Drug Resistance Nematode Intestine Anthelmintics Microscopy Pathology RNA-Seq |
author_facet |
Douglas P. Jasmer Bruce A. Rosa Rahul Tyagi Makedonka Mitreva |
author_sort |
Douglas P. Jasmer |
title |
Rapid determination of nematode cell and organ susceptibility to toxic treatments |
title_short |
Rapid determination of nematode cell and organ susceptibility to toxic treatments |
title_full |
Rapid determination of nematode cell and organ susceptibility to toxic treatments |
title_fullStr |
Rapid determination of nematode cell and organ susceptibility to toxic treatments |
title_full_unstemmed |
Rapid determination of nematode cell and organ susceptibility to toxic treatments |
title_sort |
rapid determination of nematode cell and organ susceptibility to toxic treatments |
publisher |
Elsevier |
series |
International Journal for Parasitology: Drugs and Drug Resistance |
issn |
2211-3207 |
publishDate |
2020-12-01 |
description |
In research focused on the intestine of parasitic nematodes, we recently identified small molecule inhibitors toxic to intestinal cells of larval Ascaris suum (nematode intestinal toxins/toxicants; “NITs”). Some NITs had anthelmintic activity across the phylogenetic diversity of the Nematoda. The whole-worm motility inhibition assay quantified anthelmintic activity, but worm responses to NITs in relation to pathology or affected molecular pathways was not acquired. In this study we extended this research to more comprehensively determine in whole larval A. suum the cells, organ systems, molecular targets, and potential cellular pathways involved in mechanisms of toxicity leading to cell death. The experimental system utilized fluorescent nuclear probes (bisbenzimide, propidium iodide), NITs, an A. suum larval parasite culture system and transcriptional responses (RNA-seq) to NITs. The approach provides for rapid resolution of NIT-induced cell death among organ systems (e.g. intestine, excretory, esophagus, hypodermis and seam cells, and nervous), discriminates among NITs based on cell death profiles, and identifies cells and organ systems with the greatest NIT sensitivity (e.g. intestine and apparent neuronal cells adjacent to the nerve ring). Application was extended to identify cells and organs sensitive to several existing anthelmintics. This approach also resolved intestinal cell death and irreparable damage induced in adult A. suum by two NITs, establishing a new model to elucidate relevant pathologic mechanisms in adult worms. RNA-seq analysis resolved A. suum genes responsive to treatments with three NITs, identifying dihydroorotate dehydrogenase (uridine synthesis) and RAB GTPase(s) (vesicle transport) as potential targets/pathways leading to cell death. A set of genes induced by all three NITs tested suggest common stress or survival responses activated by NITs. Beyond the presented specific lines of research, elements of the overall experimental system presented in this study have broad application toward systematic development of new anthelmintics. |
topic |
Nematode Intestine Anthelmintics Microscopy Pathology RNA-Seq |
url |
http://www.sciencedirect.com/science/article/pii/S2211320720300373 |
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