Different Stress-Induced Calcium Signatures Are Reported by Aequorin-Mediated Calcium Measurements in Living Cells of Aspergillus fumigatus.
Aspergillus fumigatus is an inhaled fungal pathogen of human lungs, the developmental growth of which is reliant upon Ca2+-mediated signalling. Ca2+ signalling has regulatory significance in all eukaryotic cells but how A. fumigatus uses intracellular Ca2+ signals to respond to stresses imposed by t...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0138008 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850162282635460608 |
|---|---|
| author | Alberto Muñoz Margherita Bertuzzi Jan Bettgenhaeuser Nino Iakobachvili Elaine M Bignell Nick D Read |
| author_facet | Alberto Muñoz Margherita Bertuzzi Jan Bettgenhaeuser Nino Iakobachvili Elaine M Bignell Nick D Read |
| author_sort | Alberto Muñoz |
| collection | DOAJ |
| description | Aspergillus fumigatus is an inhaled fungal pathogen of human lungs, the developmental growth of which is reliant upon Ca2+-mediated signalling. Ca2+ signalling has regulatory significance in all eukaryotic cells but how A. fumigatus uses intracellular Ca2+ signals to respond to stresses imposed by the mammalian lung is poorly understood. In this work, A. fumigatus strains derived from the clinical isolate CEA10, and a non-homologous recombination mutant ΔakuBKU80, were engineered to express the bioluminescent Ca2+-reporter aequorin. An aequorin-mediated method for routine Ca2+ measurements during the early stages of colony initiation was successfully developed and dynamic changes in cytosolic free calcium ([Ca2+]c) in response to extracellular stimuli were measured. The response to extracellular challenges (hypo- and hyper-osmotic shock, mechanical perturbation, high extracellular Ca2+, oxidative stress or exposure to human serum) that the fungus might be exposed to during infection, were analysed in living conidial germlings. The 'signatures' of the transient [Ca2+]c responses to extracellular stimuli were found to be dose- and age-dependent. Moreover, Ca2+-signatures associated with each physico-chemical treatment were found to be unique, suggesting the involvement of heterogeneous combinations of Ca2+-signalling components in each stress response. Concordant with the involvement of Ca2+-calmodulin complexes in these Ca2+-mediated responses, the calmodulin inhibitor trifluoperazine (TFP) induced changes in the Ca2+-signatures to all the challenges. The Ca2+-chelator BAPTA potently inhibited the initial responses to most stressors in accordance with a critical role for extracellular Ca2+ in initiating the stress responses. |
| format | Article |
| id | doaj-art-37ec36f4749d4bd7ba99833af3a7e07c |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-37ec36f4749d4bd7ba99833af3a7e07c2025-08-20T02:22:37ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01109e013800810.1371/journal.pone.0138008Different Stress-Induced Calcium Signatures Are Reported by Aequorin-Mediated Calcium Measurements in Living Cells of Aspergillus fumigatus.Alberto MuñozMargherita BertuzziJan BettgenhaeuserNino IakobachviliElaine M BignellNick D ReadAspergillus fumigatus is an inhaled fungal pathogen of human lungs, the developmental growth of which is reliant upon Ca2+-mediated signalling. Ca2+ signalling has regulatory significance in all eukaryotic cells but how A. fumigatus uses intracellular Ca2+ signals to respond to stresses imposed by the mammalian lung is poorly understood. In this work, A. fumigatus strains derived from the clinical isolate CEA10, and a non-homologous recombination mutant ΔakuBKU80, were engineered to express the bioluminescent Ca2+-reporter aequorin. An aequorin-mediated method for routine Ca2+ measurements during the early stages of colony initiation was successfully developed and dynamic changes in cytosolic free calcium ([Ca2+]c) in response to extracellular stimuli were measured. The response to extracellular challenges (hypo- and hyper-osmotic shock, mechanical perturbation, high extracellular Ca2+, oxidative stress or exposure to human serum) that the fungus might be exposed to during infection, were analysed in living conidial germlings. The 'signatures' of the transient [Ca2+]c responses to extracellular stimuli were found to be dose- and age-dependent. Moreover, Ca2+-signatures associated with each physico-chemical treatment were found to be unique, suggesting the involvement of heterogeneous combinations of Ca2+-signalling components in each stress response. Concordant with the involvement of Ca2+-calmodulin complexes in these Ca2+-mediated responses, the calmodulin inhibitor trifluoperazine (TFP) induced changes in the Ca2+-signatures to all the challenges. The Ca2+-chelator BAPTA potently inhibited the initial responses to most stressors in accordance with a critical role for extracellular Ca2+ in initiating the stress responses.https://doi.org/10.1371/journal.pone.0138008 |
| spellingShingle | Alberto Muñoz Margherita Bertuzzi Jan Bettgenhaeuser Nino Iakobachvili Elaine M Bignell Nick D Read Different Stress-Induced Calcium Signatures Are Reported by Aequorin-Mediated Calcium Measurements in Living Cells of Aspergillus fumigatus. PLoS ONE |
| title | Different Stress-Induced Calcium Signatures Are Reported by Aequorin-Mediated Calcium Measurements in Living Cells of Aspergillus fumigatus. |
| title_full | Different Stress-Induced Calcium Signatures Are Reported by Aequorin-Mediated Calcium Measurements in Living Cells of Aspergillus fumigatus. |
| title_fullStr | Different Stress-Induced Calcium Signatures Are Reported by Aequorin-Mediated Calcium Measurements in Living Cells of Aspergillus fumigatus. |
| title_full_unstemmed | Different Stress-Induced Calcium Signatures Are Reported by Aequorin-Mediated Calcium Measurements in Living Cells of Aspergillus fumigatus. |
| title_short | Different Stress-Induced Calcium Signatures Are Reported by Aequorin-Mediated Calcium Measurements in Living Cells of Aspergillus fumigatus. |
| title_sort | different stress induced calcium signatures are reported by aequorin mediated calcium measurements in living cells of aspergillus fumigatus |
| url | https://doi.org/10.1371/journal.pone.0138008 |
| work_keys_str_mv | AT albertomunoz differentstressinducedcalciumsignaturesarereportedbyaequorinmediatedcalciummeasurementsinlivingcellsofaspergillusfumigatus AT margheritabertuzzi differentstressinducedcalciumsignaturesarereportedbyaequorinmediatedcalciummeasurementsinlivingcellsofaspergillusfumigatus AT janbettgenhaeuser differentstressinducedcalciumsignaturesarereportedbyaequorinmediatedcalciummeasurementsinlivingcellsofaspergillusfumigatus AT ninoiakobachvili differentstressinducedcalciumsignaturesarereportedbyaequorinmediatedcalciummeasurementsinlivingcellsofaspergillusfumigatus AT elainembignell differentstressinducedcalciumsignaturesarereportedbyaequorinmediatedcalciummeasurementsinlivingcellsofaspergillusfumigatus AT nickdread differentstressinducedcalciumsignaturesarereportedbyaequorinmediatedcalciummeasurementsinlivingcellsofaspergillusfumigatus |