Unique Tubulin-Based Structures in the Zoonotic Apicomplexan Parasite Cryptosporidium parvum
Abstract
:1. Introduction
2. Materials and Methods
2.1. Antibody Development and Affinity-Purification
2.2. Preparation of Parasite Materials
2.3. qRT-PCR Analysis of CpTubA and CpTubB Gene Transcripts
2.4. Western Blot Analysis
2.5. Indirect Immunofluorescence Assay (IFA)
2.6. Phylogenetic Analysis of Apicomplexan α-, β- and γ-tubulin Proteins
3. Results and Discussion
3.1. The α- and β-tubulin Genes in C. Parvum Are Differentially Expressed during the Parasite Life Cycle
3.2. Tubulin Polymers Form Two Microtubular Filaments in the C. parvum Sporozoites Differing from Those in Other Apicomplexans
3.3. The Two C. Parvum Microtubules in the Sporozoite Stage Are Rigid, and the Long Microtubule Appeared to Be Associated with the Nucleus
3.4. The Two Microtubules in C. Parvum Are Restructured during the Invasion of Sporozoites and Its Transformation into Trophozoites
3.5. The Two Microtubules of C. parvum Are Present in the Asexual Developmental Stages (Merogony) and Rearranged during Parasite Development
3.6. Tubulin-Based Structures Were Present in Microgametes, but Appear Absent in Macrogametes in the Sexual Development of C. parvum
3.7. Lack of Other α- and β-tubulin Orthologs for Alternative Microtubules in the Cryptosporidium Genomes
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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Treatment Group * | Control | Colchicine (50 μM) | Paclitaxel (50 μM) | ||
---|---|---|---|---|---|
Mean ± SD (μm) | Mean ± SD (μm) | p-Value † | Mean ± SD (μm) | p-Value † | |
Long microtubule | 4.13 ± 0.41 | 4.28 ± 0.36 | 0.2079 | 4.33 ± 0.33 | 0.0683 |
Short microtubule | 1.54 ± 0.38 | 1.47 ± 0.34 | 0.6673 | 1.55 ± 0.33 | 0.9985 |
Taxonomic Group | Tubulin Subunit | ||
---|---|---|---|
Alpha | Beta | Gamma | |
Cryptosporidia | 1 | 1 | 1 |
Intestinal coccidia | 1 | 1–2 | 1 |
Cystic coccidia | 3 | 2 | 1 |
Plasmodia | 2 | 1 | 1 |
Piroplasmids | 2 | 1 | 1 |
Gregarines | 2 | 2 | 1 |
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Wang, C.; Wang, D.; Nie, J.; Gao, X.; Yin, J.; Zhu, G. Unique Tubulin-Based Structures in the Zoonotic Apicomplexan Parasite Cryptosporidium parvum. Microorganisms 2021, 9, 1921. https://doi.org/10.3390/microorganisms9091921
Wang C, Wang D, Nie J, Gao X, Yin J, Zhu G. Unique Tubulin-Based Structures in the Zoonotic Apicomplexan Parasite Cryptosporidium parvum. Microorganisms. 2021; 9(9):1921. https://doi.org/10.3390/microorganisms9091921
Chicago/Turabian StyleWang, Chenchen, Dongqiang Wang, Jiawen Nie, Xin Gao, Jigang Yin, and Guan Zhu. 2021. "Unique Tubulin-Based Structures in the Zoonotic Apicomplexan Parasite Cryptosporidium parvum" Microorganisms 9, no. 9: 1921. https://doi.org/10.3390/microorganisms9091921