Molecular responses of calcifying and non-calcifying Antarctic benthic species to Ocean Acidification - Superoxide dismutase activity
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| Main Authors: | , , , , , , , , , |
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| Format: | Dataset Open Access |
| Language: | en |
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PANGAEA
2023
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| _version_ | 1867170118209896448 |
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| author | Servetto, Natalia Ruiz, Micaela Belen Martinez, Mariano Harms, Lars de Aranzamendi, M C Alurralde, Gastón Giménez, D Abele, Doris Held, Christoph Sahade, Ricardo José |
| author_facet | Servetto, Natalia Ruiz, Micaela Belen Martinez, Mariano Harms, Lars de Aranzamendi, M C Alurralde, Gastón Giménez, D Abele, Doris Held, Christoph Sahade, Ricardo José |
| collection | Datos científicos de ciencias marinas y ambientales |
| contents | Southern Ocean organisms are thought to be particularly vulnerable to ocean acidification, as they inhabit cold waters where calcite-aragonite saturation states are naturally low. It is also generally assumed that calcifying animals would be more affected by ocean acidification than non-calcifying ones. In this context, we aimed to study the impacts of reduced pH on the ascidia Cnemidocarpa verrucosa sp. A. Here, we used gene expression profiling and enzymatic activity to study the responses of that Antarctic benthic species to ocean acidification. We sampled Cnemidocarpa verrucosa sp. A. by scuba diving at approximately 15 m depth at Carlini station, Potter Cove, King George Island, Antarctica. Caspases 3/7 activity as indicators of apoptosis intensity was measured using the Caspase-Glow 3/7 Assay kit (Promega, USA) following the manufacturer's instructions. Samples were homogenized (16-33 mg) in lysis buffer consisting in 25 mM HEPES, 5 mM MgCl₂·6H₂O, 1 mM EGTA, 1 μg/mL pepstatin, 1 μg/mL leupectin, and 1 μg/mL aprotinin at a ratio 1:100 (Rivera-Ingraham et al., 2013) using a Precellys homogenizer (2 cycles at 5,500 x g at 4°C for 20 s). Homogenates were centrifuged at 13,000 x g at 4°C for 15 min and the supernatant was used to measure luminescence using Tristar LB941 plate reader (Berthold Technologies GmbH & Co. KG, Bad Wildbad, Germany). The total protein content of the samples was measured using the method of Bradford (1976). Caspase/Apoptotic activity was expressed as relative light units (RLU) per μg of protein × 104. |
| format | Dataset Open Access |
| id | pangaea_https___doi_org_10_1594_PANGAEA_956185 |
| institution | PANGAEA |
| language | en |
| publishDate | 2023 |
| publisher | PANGAEA |
| record_format | pangaea |
| spellingShingle | Molecular responses of calcifying and non-calcifying Antarctic benthic species to Ocean Acidification - Superoxide dismutase activity Servetto, Natalia Ruiz, Micaela Belen Martinez, Mariano Harms, Lars de Aranzamendi, M C Alurralde, Gastón Giménez, D Abele, Doris Held, Christoph Sahade, Ricardo José Ant_PotterCove_2015; Antarctica; apoptosis; Bradford method (1976); Buffer; Calculated; Calculated average/mean values; Caspase; Change of extinction; Cnemidocarpa verrucosa, superoxide dismutase, in extract; Cnemidocarpa verrucosa, superoxide dismutase, per wet mass; Cnemidocarpa verrucosa, superoxide dismutase, unit per protein mass; Cnemidocarpa verrucosa sp. A; Date/time end, experiment; Date/time start, experiment; DIVER; Event label; laboratory study; Potter Cove; Potter Cove, King George Island, Antarctic Peninsula; Proteins; Sample, wet mass; Sample code/label; Sample volume; Sampling by diver; Species; Spectrophotometer UV/Vis, Beckman Coulter, DU800; Temperature, water; Treatment; Tunicata; Type of study Southern Ocean organisms are thought to be particularly vulnerable to ocean acidification, as they inhabit cold waters where calcite-aragonite saturation states are naturally low. It is also generally assumed that calcifying animals would be more affected by ocean acidification than non-calcifying ones. In this context, we aimed to study the impacts of reduced pH on the ascidia Cnemidocarpa verrucosa sp. A. Here, we used gene expression profiling and enzymatic activity to study the responses of that Antarctic benthic species to ocean acidification. We sampled Cnemidocarpa verrucosa sp. A. by scuba diving at approximately 15 m depth at Carlini station, Potter Cove, King George Island, Antarctica. Caspases 3/7 activity as indicators of apoptosis intensity was measured using the Caspase-Glow 3/7 Assay kit (Promega, USA) following the manufacturer's instructions. Samples were homogenized (16-33 mg) in lysis buffer consisting in 25 mM HEPES, 5 mM MgCl₂·6H₂O, 1 mM EGTA, 1 μg/mL pepstatin, 1 μg/mL leupectin, and 1 μg/mL aprotinin at a ratio 1:100 (Rivera-Ingraham et al., 2013) using a Precellys homogenizer (2 cycles at 5,500 x g at 4°C for 20 s). Homogenates were centrifuged at 13,000 x g at 4°C for 15 min and the supernatant was used to measure luminescence using Tristar LB941 plate reader (Berthold Technologies GmbH & Co. KG, Bad Wildbad, Germany). The total protein content of the samples was measured using the method of Bradford (1976). Caspase/Apoptotic activity was expressed as relative light units (RLU) per μg of protein × 104. |
| title | Molecular responses of calcifying and non-calcifying Antarctic benthic species to Ocean Acidification - Superoxide dismutase activity |
| topic | Ant_PotterCove_2015; Antarctica; apoptosis; Bradford method (1976); Buffer; Calculated; Calculated average/mean values; Caspase; Change of extinction; Cnemidocarpa verrucosa, superoxide dismutase, in extract; Cnemidocarpa verrucosa, superoxide dismutase, per wet mass; Cnemidocarpa verrucosa, superoxide dismutase, unit per protein mass; Cnemidocarpa verrucosa sp. A; Date/time end, experiment; Date/time start, experiment; DIVER; Event label; laboratory study; Potter Cove; Potter Cove, King George Island, Antarctic Peninsula; Proteins; Sample, wet mass; Sample code/label; Sample volume; Sampling by diver; Species; Spectrophotometer UV/Vis, Beckman Coulter, DU800; Temperature, water; Treatment; Tunicata; Type of study |
| url | https://doi.org/10.1594/PANGAEA.956185 |