{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T12:17:47Z","timestamp":1792844267629,"version":"4.5.0"},"reference-count":36,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,9,10]],"date-time":"2021-09-10T00:00:00Z","timestamp":1631232000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National key R&amp;D Program of China","award":["2017YFC1601200"],"award-info":[{"award-number":["2017YFC1601200"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Cryptosporidium parasites are known to be highly divergent from other apicomplexan species at evolutionary and biological levels. Here we provide evidence showing that the zoonotic Cryptosporidium parvum also differs from other apicomplexans, such as Toxoplasma gondii, by possessing only two tubulin-based filamentous structures, rather than an array of subpellicular microtubules. Using an affinity-purified polyclonal antibody against C. parvum \u03b2-tubulin (CpTubB), we observed a long and a short microtubule that are rigid and stable in the sporozoites and restructured during the intracellular parasite development. In asexual development (merogony), the two restructuring microtubules are present in pairs (one pair per nucleus or merozoites). In sexual developmental stages, tubulin-based structures are detectable only in microgametes, but undetectable in macrogametes. These observations indicate that C. parvum parasites use unique microtubule structures that differ from other apicomplexans as part of their cytoskeletal elements.<\/jats:p>","DOI":"10.3390\/microorganisms9091921","type":"journal-article","created":{"date-parts":[[2021,9,12]],"date-time":"2021-09-12T21:45:57Z","timestamp":1631483157000},"page":"1921","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Unique Tubulin-Based Structures in the Zoonotic Apicomplexan Parasite Cryptosporidium parvum"],"prefix":"10.3390","volume":"9","author":[{"given":"Chenchen","family":"Wang","sequence":"first","affiliation":[{"name":"Key Laboratory for Zoonoses Research of the Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dongqiang","family":"Wang","sequence":"additional","affiliation":[{"name":"Key Laboratory for Zoonoses Research of the Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiawen","family":"Nie","sequence":"additional","affiliation":[{"name":"Key Laboratory for Zoonoses Research of the Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Gao","sequence":"additional","affiliation":[{"name":"Key Laboratory for Zoonoses Research of the Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jigang","family":"Yin","sequence":"additional","affiliation":[{"name":"Key Laboratory for Zoonoses Research of the Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3888-0659","authenticated-orcid":false,"given":"Guan","family":"Zhu","sequence":"additional","affiliation":[{"name":"Key Laboratory for Zoonoses Research of the Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,9,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Goodson, H.V., and Jonasson, E.M. (2018). Microtubules and Microtubule-Associated Proteins. Cold Spring Harb Perspect. Biol., 10.","DOI":"10.1101\/cshperspect.a022608"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1240","DOI":"10.1177\/1535370219867296","article-title":"Microtubules: Evolving roles and critical cellular interactions","volume":"244","author":"Logan","year":"2019","journal-title":"Exp. Biol. Med. (Maywood)"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e56635","DOI":"10.7554\/eLife.56635","article-title":"Essential function of the alveolin network in the subpellicular microtubules and conoid assembly in Toxoplasma gondii","volume":"9","author":"Tosetti","year":"2020","journal-title":"Elife"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"979","DOI":"10.1016\/j.pt.2020.09.001","article-title":"The Riveting Cellular Structures of Apicomplexan Parasites","volume":"36","author":"Harding","year":"2020","journal-title":"Trends Parasitol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"688","DOI":"10.1016\/j.pt.2020.05.001","article-title":"Evolution, Composition, Assembly, and Function of the Conoid in Apicomplexa","volume":"36","author":"Tosetti","year":"2020","journal-title":"Trends Parasitol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1128\/MMBR.66.1.21-38.2002","article-title":"Cytoskeleton of apicomplexan parasites","volume":"66","author":"Morrissette","year":"2002","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1038\/nrmicro.2017.86","article-title":"Gliding motility powers invasion and egress in Apicomplexa","volume":"15","author":"Frenal","year":"2017","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1186\/s44149-021-00002-y","article-title":"Current status and challenges in drug discovery against the globally important zoonotic cryptosporidiosis","volume":"1","author":"Zhu","year":"2021","journal-title":"Anim. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1007\/s40588-021-00159-7","article-title":"Cryptosporidium: Host-Parasite Interactions and Pathogenesis","volume":"8","author":"Pinto","year":"2021","journal-title":"Curr. Clin. Microbiol. Rep."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1016\/j.pt.2019.12.016","article-title":"A One Health Approach to Tackle Cryptosporidiosis","volume":"36","author":"Innes","year":"2020","journal-title":"Trends Parasitol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1111\/pim.12350","article-title":"Cryptosporidium in humans and animals-a one health approach to prophylaxis","volume":"38","author":"Ryan","year":"2016","journal-title":"Parasite Immunol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1007\/BF00536023","article-title":"Ultrastructure of Cryptosporidium muris (strain RN 66) parasitizing the murine stomach","volume":"74","author":"Uni","year":"1987","journal-title":"Parasitol. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1017\/S003118200700385X","article-title":"Electron microscopic observation of cytoskeletal frame structures and detection of tubulin on the apical region of Cryptosporidium parvum sporozoites","volume":"135","author":"Matsubayashi","year":"2008","journal-title":"Parasitology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"995","DOI":"10.1645\/GE-495R.1","article-title":"Distribution of Cryptosporidium parvum sporozoite apical organelles during attachment to and internalization by cultured biliary epithelial cells","volume":"91","author":"Huang","year":"2005","journal-title":"J. Parasitol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1110","DOI":"10.1093\/infdis\/jix689","article-title":"The Existing Drug Vorinostat as a New Lead Against Cryptosporidiosis by Targeting the Parasite Histone Deacetylases","volume":"217","author":"Guo","year":"2018","journal-title":"J. Infect. Dis."},{"key":"ref_16","first-page":"173","article-title":"An improved protocol for coupling synthetic peptides to carrier proteins for antibody production using DMF to solubilize peptides","volume":"18","author":"Lateef","year":"2007","journal-title":"J. Biomol. Tech."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Greenfield, E.A. (2020). Standard Immunization of Rabbits. Cold Spring Harb Protoc., 2020.","DOI":"10.1101\/pdb.prot100305"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1007\/978-1-59745-542-8_22","article-title":"Affinity purification of autoantibodies from an antigen strip excised from a nitrocellulose protein blot","volume":"536","author":"Kurien","year":"2009","journal-title":"Methods Mol. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1016\/j.ijpara.2006.02.023","article-title":"Quantitative and qualitative comparisons of Cryptosporidium faecal purification procedures for the isolation of oocysts suitable for proteomic analysis","volume":"36","author":"Truong","year":"2006","journal-title":"Int. J. Parasitol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1007\/978-1-4939-9748-0_18","article-title":"High-Throughput Screening of Drugs against the Growth of Cryptosporidium parvum In Vitro by qRT-PCR","volume":"2052","author":"Zhang","year":"2020","journal-title":"Methods Mol. Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1753","DOI":"10.1128\/IAI.05862-11","article-title":"Involvement of host cell integrin alpha2 in Cryptosporidium parvum infection","volume":"80","author":"Zhang","year":"2012","journal-title":"Infect. Immun."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Zhang, H., and Zhu, G. (2015). Quantitative RT-PCR assay for high-throughput screening (HTS) of drugs against the growth of Cryptosporidium parvum in vitro. Front. Microbiol., 6.","DOI":"10.3389\/fmicb.2015.00991"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Mauzy, M.J., Enomoto, S., Lancto, C.A., Abrahamsen, M.S., and Rutherford, M.S. (2012). The Cryptosporidium parvum transcriptome during in vitro development. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0031715"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1111\/jeu.12272","article-title":"Differential Gene Expression and Protein Localization of Cryptosporidium parvum Fatty Acyl-CoA Synthetase Isoforms","volume":"63","author":"Guo","year":"2016","journal-title":"J. Eukaryot. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Zhang, H., Guo, F., and Zhu, G. (2015). Cryptosporidium Lactate Dehydrogenase Is Associated with the Parasitophorous Vacuole Membrane and Is a Potential Target for Developing Therapeutics. PLoS Pathog., 11.","DOI":"10.1371\/journal.ppat.1005250"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Zhang, H., Guo, F., Zhou, H., and Zhu, G. (2012). Transcriptome analysis reveals unique metabolic features in the Cryptosporidium parvum Oocysts associated with environmental survival and stresses. BMC Genom., 13.","DOI":"10.1186\/1471-2164-13-647"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1083\/jcb.200112086","article-title":"A novel polymer of tubulin forms the conoid of Toxoplasma gondii","volume":"156","author":"Hu","year":"2002","journal-title":"J. Cell Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1021\/pr900699a","article-title":"Post-translational modifications to Toxoplasma gondii alpha- and beta-tubulins include novel C-terminal methylation","volume":"9","author":"Xiao","year":"2010","journal-title":"J. Proteome Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1407","DOI":"10.1016\/j.chom.2021.07.002","article-title":"Cryptosporidium rhoptry effector protein ROP1 injected during invasion targets the host cytoskeletal modulator LMO7","volume":"29","author":"Guerin","year":"2021","journal-title":"Cell Host Microbe"},{"key":"ref_30","first-page":"2795","article-title":"Functional coupling of microtubules to membranes\u2014Implications for membrane structure and dynamics","volume":"125","author":"Stephens","year":"2012","journal-title":"J. Cell Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"a025817","DOI":"10.1101\/cshperspect.a025817","article-title":"Microtubule-Based Transport and the Distribution, Tethering, and Organization of Organelles","volume":"9","author":"Barlan","year":"2017","journal-title":"Cold Spring Harb Perspect Biol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Gallardo, P., Barrales, R.R., Daga, R.R., and Salas-Pino, S. (2019). Nuclear Mechanics in the Fission Yeast. Cells, 8.","DOI":"10.3390\/cells8101285"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1016\/j.ceb.2020.03.001","article-title":"The role of the cell nucleus in mechanotransduction","volume":"63","author":"Janota","year":"2020","journal-title":"Curr. Opin. Cell Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1038\/nrmicro3184","article-title":"Cell division in apicomplexan parasites","volume":"12","author":"Francia","year":"2014","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2226","DOI":"10.1038\/s41564-019-0539-x","article-title":"Life cycle progression and sexual development of the apicomplexan parasite Cryptosporidium parvum","volume":"4","author":"Tandel","year":"2019","journal-title":"Nat. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1128\/EC.00225-14","article-title":"Targeting Toxoplasma tubules: Tubulin, microtubules, and associated proteins in a human pathogen","volume":"14","author":"Morrissette","year":"2015","journal-title":"Eukaryot. Cell"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/9\/9\/1921\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:00:06Z","timestamp":1760166006000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/9\/9\/1921"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,9,10]]},"references-count":36,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2021,9]]}},"alternative-id":["microorganisms9091921"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms9091921","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,9,10]]}}}