TY - JOUR
T1 - Dramatic changes in gene expression in different forms of Crithidia fasciculata reveal potential mechanisms for insect-specific adhesion in kinetoplastid parasites
AU - Filosa, John N.
AU - Berry, Corbett T.
AU - Ruthel, Gordon
AU - Beverley, Stephen M.
AU - Warren, Wesley C.
AU - Tomlinson, Chad
AU - Myler, Peter J.
AU - Dudkin, Elizabeth A.
AU - Povelones, Megan L.
AU - Povelones, Michael
N1 - Funding Information:
This work was supported by the National Science Foundation under grant number MCB-1651517 to MLP. https://www.nsf.gov. The PennVet Imaging Core (GR) is supported by a National Institutes of Health Shared Instrument Grant S10 OD021633-01 to Bruce Freedman. This study was financially supported in part by NIH grant AI29646 to SMB, NIH-NHGRI grant 5U54HG00307907 to Richard K. Wilson, Director of the Genome Institute at Washington University, and NIH-NIAID grant AI103858 to PJM. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors thank Emmanual Tetaud for providing the pNUS plasmids. We acknowledge Natalia S. Akopyants and George Lye, who prepared and performed DNA quality control (Washington University, St. Louis) for genome sequencing. We thank Gowthaman Ramasamy and Sacha Steinbiss for annotation of the C. fasciculata genome. We thank Ana Misic for help with library preparation for RNAseq, Daniel Beiting and the Center for Host-Microbial Interactions for help with RNAseq data generation and analysis, and Henry Neeb for assistance with R. We thank David S. Roos for input on the project. We thank Omar S. Harb for help in obtaining the KEGG gene sets from TriTrypDB. Thanks to Madeline Malfara, John DiMaio, and Michael Yoder for helpful comments on the manuscript. We thank Elizabeth B. Edgerton for help with mosquito rearing and maintenance.
Publisher Copyright:
© 2019 Filosa et al.
PY - 2019/7
Y1 - 2019/7
N2 - Kinetoplastids are a group of parasites that includes several medically-important species. These human-infective species are transmitted by insect vectors in which the parasites undergo specific developmental transformations. For each species, this includes a stage in which parasites adhere to insect tissue via a hemidesmosome-like structure. Although this structure has been described morphologically, it has never been molecularly characterized. We are using Crithidia fasciculata, an insect parasite that produces large numbers of adherent parasites inside its mosquito host, as a model kinetoplastid to investigate both the mechanism of adherence and the signals required for differentiation to an adherent form. An advantage of C. fasciculata is that adherent parasites can be generated both in vitro, allowing a direct comparison to cultured swimming forms, as well as in vivo within the mosquito. Using RNAseq, we identify genes associated with adherence in C. fasciculata. As almost all of these genes have orthologs in other kinetoplastid species, our findings may reveal shared mechanisms of adherence, allowing investigation of a crucial step in parasite development and disease transmission. In addition, dual-RNAseq allowed us to explore the interaction between the parasites and the mosquito. Although the infection is well-tolerated, anti-microbial peptides and other components of the mosquito innate immune system are upregulated. Our findings indicate that C. fasciculata is a powerful model system for probing kinetoplastid-insect interactions.
AB - Kinetoplastids are a group of parasites that includes several medically-important species. These human-infective species are transmitted by insect vectors in which the parasites undergo specific developmental transformations. For each species, this includes a stage in which parasites adhere to insect tissue via a hemidesmosome-like structure. Although this structure has been described morphologically, it has never been molecularly characterized. We are using Crithidia fasciculata, an insect parasite that produces large numbers of adherent parasites inside its mosquito host, as a model kinetoplastid to investigate both the mechanism of adherence and the signals required for differentiation to an adherent form. An advantage of C. fasciculata is that adherent parasites can be generated both in vitro, allowing a direct comparison to cultured swimming forms, as well as in vivo within the mosquito. Using RNAseq, we identify genes associated with adherence in C. fasciculata. As almost all of these genes have orthologs in other kinetoplastid species, our findings may reveal shared mechanisms of adherence, allowing investigation of a crucial step in parasite development and disease transmission. In addition, dual-RNAseq allowed us to explore the interaction between the parasites and the mosquito. Although the infection is well-tolerated, anti-microbial peptides and other components of the mosquito innate immune system are upregulated. Our findings indicate that C. fasciculata is a powerful model system for probing kinetoplastid-insect interactions.
UR - http://www.scopus.com/inward/record.url?scp=85071348664&partnerID=8YFLogxK
U2 - 10.1371/journal.pntd.0007570
DO - 10.1371/journal.pntd.0007570
M3 - Article
C2 - 31356610
AN - SCOPUS:85071348664
SN - 1935-2727
VL - 13
JO - PLoS Neglected Tropical Diseases
JF - PLoS Neglected Tropical Diseases
IS - 7
M1 - e0007570
ER -