A fast and cost-effective microsampling protocol incorporating reduced animal usage for time-series transcriptomics in rodent malaria parasites
Abstract Background The transcriptional regulation that occurs in malaria parasites during the erythrocytic stages of infection can be studied in vivo with rodent malaria parasites propagated in mice. Time-series transcriptome profiling commonly involves the euthanasia of groups of mice at specific...
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ftdoajarticles:oai:doaj.org/article:76ee734b6c2e43039524fbe5e0e76d9e 2023-05-15T15:09:35+02:00 A fast and cost-effective microsampling protocol incorporating reduced animal usage for time-series transcriptomics in rodent malaria parasites Abhinay Ramaprasad Amit Kumar Subudhi Richard Culleton Arnab Pain 2019-01-01T00:00:00Z https://doi.org/10.1186/s12936-019-2659-4 https://doaj.org/article/76ee734b6c2e43039524fbe5e0e76d9e EN eng BMC http://link.springer.com/article/10.1186/s12936-019-2659-4 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-019-2659-4 1475-2875 https://doaj.org/article/76ee734b6c2e43039524fbe5e0e76d9e Malaria Journal, Vol 18, Iss 1, Pp 1-11 (2019) Rodent malaria parasites Transcriptomics Time-series Plasmodium Malaria Microsampling Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2019 ftdoajarticles https://doi.org/10.1186/s12936-019-2659-4 2023-01-08T01:28:31Z Abstract Background The transcriptional regulation that occurs in malaria parasites during the erythrocytic stages of infection can be studied in vivo with rodent malaria parasites propagated in mice. Time-series transcriptome profiling commonly involves the euthanasia of groups of mice at specific time points followed by the extraction of parasite RNA from whole blood samples. Current methodologies for parasite RNA extraction involve several steps and when multiple time points are profiled, these protocols are laborious, time-consuming, and require the euthanization of large cohorts of mice. Results A simplified protocol has been designed for parasite RNA extraction from blood volumes as low as 20 μL (microsamples), serially bled from mice via tail snips and directly lysed with TRIzol reagent. Gene expression data derived from microsampling using RNA-seq were closely matched to those derived from larger volumes of leucocyte-depleted and saponin-treated blood obtained from euthanized mice with high reproducibility between biological replicates. Transcriptome profiling of microsamples taken at different time points during the intra-erythrocytic developmental cycle of the rodent malaria parasite Plasmodium vinckei revealed the transcriptional cascade commonly observed in malaria parasites. Conclusions Microsampling is a quick, robust and cost-efficient approach to sample collection for in vivo time-series transcriptomic studies in rodent malaria parasites. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 18 1 |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Rodent malaria parasites Transcriptomics Time-series Plasmodium Malaria Microsampling Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
spellingShingle |
Rodent malaria parasites Transcriptomics Time-series Plasmodium Malaria Microsampling Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Abhinay Ramaprasad Amit Kumar Subudhi Richard Culleton Arnab Pain A fast and cost-effective microsampling protocol incorporating reduced animal usage for time-series transcriptomics in rodent malaria parasites |
topic_facet |
Rodent malaria parasites Transcriptomics Time-series Plasmodium Malaria Microsampling Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background The transcriptional regulation that occurs in malaria parasites during the erythrocytic stages of infection can be studied in vivo with rodent malaria parasites propagated in mice. Time-series transcriptome profiling commonly involves the euthanasia of groups of mice at specific time points followed by the extraction of parasite RNA from whole blood samples. Current methodologies for parasite RNA extraction involve several steps and when multiple time points are profiled, these protocols are laborious, time-consuming, and require the euthanization of large cohorts of mice. Results A simplified protocol has been designed for parasite RNA extraction from blood volumes as low as 20 μL (microsamples), serially bled from mice via tail snips and directly lysed with TRIzol reagent. Gene expression data derived from microsampling using RNA-seq were closely matched to those derived from larger volumes of leucocyte-depleted and saponin-treated blood obtained from euthanized mice with high reproducibility between biological replicates. Transcriptome profiling of microsamples taken at different time points during the intra-erythrocytic developmental cycle of the rodent malaria parasite Plasmodium vinckei revealed the transcriptional cascade commonly observed in malaria parasites. Conclusions Microsampling is a quick, robust and cost-efficient approach to sample collection for in vivo time-series transcriptomic studies in rodent malaria parasites. |
format |
Article in Journal/Newspaper |
author |
Abhinay Ramaprasad Amit Kumar Subudhi Richard Culleton Arnab Pain |
author_facet |
Abhinay Ramaprasad Amit Kumar Subudhi Richard Culleton Arnab Pain |
author_sort |
Abhinay Ramaprasad |
title |
A fast and cost-effective microsampling protocol incorporating reduced animal usage for time-series transcriptomics in rodent malaria parasites |
title_short |
A fast and cost-effective microsampling protocol incorporating reduced animal usage for time-series transcriptomics in rodent malaria parasites |
title_full |
A fast and cost-effective microsampling protocol incorporating reduced animal usage for time-series transcriptomics in rodent malaria parasites |
title_fullStr |
A fast and cost-effective microsampling protocol incorporating reduced animal usage for time-series transcriptomics in rodent malaria parasites |
title_full_unstemmed |
A fast and cost-effective microsampling protocol incorporating reduced animal usage for time-series transcriptomics in rodent malaria parasites |
title_sort |
fast and cost-effective microsampling protocol incorporating reduced animal usage for time-series transcriptomics in rodent malaria parasites |
publisher |
BMC |
publishDate |
2019 |
url |
https://doi.org/10.1186/s12936-019-2659-4 https://doaj.org/article/76ee734b6c2e43039524fbe5e0e76d9e |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 18, Iss 1, Pp 1-11 (2019) |
op_relation |
http://link.springer.com/article/10.1186/s12936-019-2659-4 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-019-2659-4 1475-2875 https://doaj.org/article/76ee734b6c2e43039524fbe5e0e76d9e |
op_doi |
https://doi.org/10.1186/s12936-019-2659-4 |
container_title |
Malaria Journal |
container_volume |
18 |
container_issue |
1 |
_version_ |
1766340749633257472 |