Basement membrane proteins as a substrate for efficient Trypanosoma brucei differentiation in vitro
| العنوان: | Basement membrane proteins as a substrate for efficient Trypanosoma brucei differentiation in vitro |
|---|---|
| المؤلفون: | Mathieu Cayla, Federico Rojas, Keith R. Matthews |
| المصدر: | Rojas, F, Cayla, M & Matthews, K R 2021, ' Basement membrane proteins as a substrate for efficient Trypanosoma brucei differentiation in vitro ', PLoS Neglected Tropical Diseases, vol. 15, no. 4 . https://doi.org/10.1371/journal.pntd.0009284 PLoS Neglected Tropical Diseases, Vol 15, Iss 4, p e0009284 (2021) PLoS Neglected Tropical Diseases |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | 0301 basic medicine, Life Cycles, Cellular differentiation, RC955-962, Protozoan Proteins, Matrix (biology), Biochemistry, Parasitic Cell Cycles, Basement Membrane, Extracellular matrix, 0302 clinical medicine, Arctic medicine. Tropical medicine, Protozoans, 0303 health sciences, biology, Chemistry, Cell Cycle, 030302 biochemistry & molecular biology, Eukaryota, Quorum Sensing, Cell Differentiation, Proteases, Cell cycle, Enzymes, Extracellular Matrix, Cell biology, Infectious Diseases, medicine.anatomical_structure, Cellular Structures and Organelles, Public aspects of medicine, RA1-1270, Research Article, Trypanosoma, Parasitic Life Cycles, Trypanosoma brucei brucei, Virulence, Trypanosoma brucei, 03 medical and health sciences, Trypanosoma Brucei, medicine, Animals, Secretion, 030304 developmental biology, Basement membrane, Life Cycle Stages, Organisms, Public Health, Environmental and Occupational Health, Biology and Life Sciences, Proteins, Cell Biology, biology.organism_classification, Parasitic Protozoans, In vitro, Culture Media, 030104 developmental biology, Enzymology, Parasitology, Collagens, 030217 neurology & neurosurgery, Developmental Biology, Trypanosoma Brucei Gambiense |
| الوصف: | The ability to reproduce the developmental events of trypanosomes that occur in their mammalian host in vitro offers significant potential to assist in understanding of the underlying biology of the process. For example, the transition from bloodstream slender to bloodstream stumpy forms is a quorum-sensing response to the parasite-derived peptidase digestion products of environmental proteins. As an abundant physiological substrate in vivo, we studied the ability of a basement membrane matrix enriched gel (BME) in the culture medium to support differentiation of pleomorphic Trypanosoma brucei to stumpy forms. BME comprises extracellular matrix proteins, which are among the most abundant proteins found in connective tissues in mammals and known substrates of parasite-released peptidases. We previously showed that two of these released peptidases are involved in generating a signal that promotes slender-to-stumpy differentiation. Here, we tested the ability of basement membrane extract to enhance parasite differentiation through its provision of suitable substrates to generate the quorum sensing signal, namely oligopeptides. Our results show that when grown in the presence of BME, T. brucei pleomorphic cells arrest at the G0/1 phase of the cell cycle and express the differentiation marker PAD1, the response being restricted to differentiation-competent parasites. Further, the stumpy forms generated in BME medium are able to efficiently proceed onto the next life cycle stage in vitro, procyclic forms, when incubated with cis-aconitate, further validating the in vitro BME differentiation system. Hence, BME provides a suitable in vitro substrate able to accurately recapitulate physiological parasite differentiation without the use of experimental animals. Author summary African trypanosome parasites responsible for human and animal trypanosomiasis, live extracellularly in their mammalian host and exploit environmental information to regulate their virulence and transmissibility. Morphological slender forms that proliferate in the mammalian host transform into the non-proliferative stumpy forms by receiving an extracellular signal generated by the accumulating parasites. Parasites secrete peptidases into the host that degrade extracellular matrix (ECM) proteins. In this study, we evaluated the capacity to generate stumpy forms in vitro by including a basement membrane extract into the culture medium. We show that cells exposed to ECM proteins are able to differentiate efficiently into cell-cycle arrested stumpy forms and that the generated differentiation signal depends on the stumpy induction signalling pathway. This method can be used to generate stumpy forms in vitro, these being suitable for experimental analysis, thereby reducing the use of mice. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d44c5e74428dac6efd24020a69b1d0c0 https://www.pure.ed.ac.uk/ws/files/209116413/journal.pntd.0009284.pdf |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....d44c5e74428dac6efd24020a69b1d0c0 |
| قاعدة البيانات: | OpenAIRE |
| الوصف غير متاح. |